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   2017| July-September  | Volume 13 | Issue 51  
    Online since October 11, 2017

 
 
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ORIGINAL ARTICLES
Bioactive constituents, radical scavenging, and antibacterial properties of the leaves and stem essential oils from Peperomia pellucida (L.) kunth
Sunday O Okoh, Benson C Iweriebor, Omobola O Okoh, Anthony I Okoh
July-September 2017, 13(51):392-400
DOI:10.4103/pm.pm_106_17  PMID:29142389
Background: Peperomia pellucida is an annual herbaceous ethnomedicinal plant used in the treatment of a variety of communicable and noncommunicable diseases in the Amazon region. Objective: The study aimed at profiling the bioactive constituents of the leaves and stem essential oils (LEO and SEO) of P. pellucida, their in vitro antibacterial and radical scavenging properties as probable lead constituents in the management of oxidative stress and infectious diseases. Materials and Methods: The EOs were obtained from the leaves and stem P. pellucida using modified Clevenger apparatus and characterized by a high-resolution gas chromatography-mass spectrometry, while the radicals scavenging and antibacterial effects on four oxidants and six reference bacteria strains were examined by spectrophotometric and agar diffusion techniques, respectively. Results: The EOs exhibited strong antibacterial activities against six bacteria (Escherichia coli [180], Enterobacter cloacae, Mycobacterium smegmatis, Listeria ivanovii, Staphylococcus aureus, Streptococcus uberis, and Vibrio paraheamolyticus) strains. The SEO antibacterial activities were not significantly different (P < 0.05) from the LEO against most of the test bacteria with minimum inhibitory concentration ranging between 0.15 and 0.20 mg/mL for both EOs. The two oils were bactericidal at 0.20 mg/mL against S. aureus while the minimum bactericidal concentration (0.15 mg/mL) of LEO against L. ivanovii was lower than of SEO (0.20 mg/mL) after 24 h. The LEO IC50value (1.67 mg/mL) revealed more radical scavenging activity than the SEO (2.83 mg/mL) and reference compounds against 2,2-diphenyl-1-picrylhydrazyl radical. The EOs also scavenged three other different radicals (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical , lipid peroxyl radical, and nitric oxide radical) in concentration-dependent manner. Conclusion: Our results suggest that apart from the indigenous uses of the plant extracts, the EO contains strong bioactive compounds with antibacterial and radicals scavenging properties and may be good alternative candidates in the search for novel potent antibiotics in this present era of increasing multidrug-resistant bacterial strains as well as effective antioxidants agents. Abbreviations used: GC-MS: Gas chromatography-mass spectrometry, DPPH: 2,2-diphenyl-1-picrylhydrazyl, ABTS: 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, DMSO: Dimethyl sulfoxide, LP: Lipid peroxide radical, NO: Nitric oxide radical, LEO: Leaf essential oil, SEO: Stem essential oil, RC: Reference compound, TBARS: Thiobarbituric acid
  4,804 275 5
Chemical composition of Moringa oleifera ethyl acetate fraction and its biological activity in diabetic human dermal fibroblasts
Sivapragasam Gothai, Katyakyini Muniandy, Mazni Abu Zarin, Tan Woan Sean, S Suresh Kumar, Murugan A Munusamy, Sharida Fakurazi, Palanisamy Arulselvan
July-September 2017, 13(51):462-469
DOI:10.4103/pm.pm_368_16  PMID:29142400
Background: Moringa oleifera (MO), commonly known as the drumstick tree, is used in folklore medicine for the treatment of skin disease. Objective: The objective of this study is to evaluate the ethyl acetate (EtOAc) fraction of MO leaves for in vitro antibacterial, antioxidant, and wound healing activities and conduct gas chromatography-mass spectrometry (GC-MS) analysis. Materials and Methods: Antibacterial activity was evaluated against six Gram-positive bacteria and 10 Gram-negative bacteria by disc diffusion method. Free radical scavenging activity was assessed by 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radical hydrogen peroxide scavenging and total phenolic content (TPC). Wound healing efficiency was studied using cell viability, proliferation, and scratch assays in diabetic human dermal fibroblast (HDF-D) cells. Results: The EtOAc fraction showed moderate activity against all bacterial strains tested, and the maximum inhibition zone was observed against Streptococcus pyogenes (30 mm in diameter). The fraction showed higher sensitivity to Gram-positive strains than Gram-negative strains. In the quantitative analysis of antioxidant content, the EtOAc fraction was found to have a TPC of 65.81 ± 0.01. The DPPH scavenging activity and the hydrogen peroxide assay were correlated with the TPC value, with IC50values of 18.21 ± 0.06 and 59.22 ± 0.04, respectively. The wound healing experiment revealed a significant enhancement of cell proliferation and migration of HDF-D cells. GC-MS analysis confirmed the presence of 17 bioactive constituents that may be the principal factors in the significant antibacterial, antioxidant, and wound healing activity. Conclusion: The EtOAc fraction of MO leaves possesses remarkable wound healing properties, which can be attributed to the antibacterial and antioxidant activities of the fraction. Abbreviations used: MO: Moringa oleifera; EtOAc: Ethyl acetate; GC-MS: Gas Chromatography-Mass Spectrometry; HDF-D: Diabetic Human Dermal Fibroblast cells.
  4,399 276 -
Effects of Plantago major extracts and its chemical compounds on proliferation of cancer cells and cytokines production of lipopolysaccharide-activated THP-1 macrophages
Kartini , Suratsawadee Piyaviriyakul, Suchitra Thongpraditchote, Pongpun Siripong, Omboon Vallisuta
July-September 2017, 13(51):393-399
DOI:10.4103/pm.pm_406_16  PMID:28839362
Background: Plantago major has been reported to have anticancer and anti-inflammatory properties. However, its antiproliferative and anti-inflammatory mechanisms have not been fully elucidated. Moreover, which plant parts are more suitable as starting materials has not been explored. Objectives: To investigate the antiproliferative activity of P. major extracts against MCF-7, MDA-MB-231, HeLaS3, A549, and KB cancer cell lines as well as their effects on inflammatory cytokines (tumor necrosis factor [TNF]-a, interleukin [IL]-1β, IL-6, and interferon [IFN]-g) production by lipopolysaccharide (LPS)-stimulated THP-1 macrophages. Materials and Methods: The methanol and aqueous extracts of P. major from different plant parts and its chemical compounds, i.e., ursolic acid (UA), oleanolic acid (OA), and aucubin were tested in this experiment. Results: Methanol and aqueous extracts of P. major seeds exhibited the greatest antiproliferative activity. The methanol extracts of seeds also demonstrated the highest inhibition of TNF-a, IL-1β, IL-6, and IFN-g production. Interestingly, the roots, which were commonly discarded, exhibited comparable activities to those of leaves and petioles. Furthermore, UA exhibited stronger activities than OA and aucubin. Conclusions: The seeds are being proposed as the main source for further development of anticancer and anti-inflammatory products, whereas the roots could be included in the preparation of P. major derived products with respect to anti-inflammatory. Abbreviations used: TNF: Tumor Necrosis Factor; IL: Interleukin; IFN: Interferon; HPTLC: High Performance Thin Layer Chromatography; UA: Ursolic Acid; OA: Oleanolic Acid; AUC: Aucubin.
  3,918 245 7
Analysis of free amino acids in different extracts of Orthosiphon stamineus leaves by high-performance liquid chromatography combined with solid-phase extraction
Armaghan Shafaei, Nor Hidayah Ab Halim, Norhidayah Zakaria, Zhari Ismail
July-September 2017, 13(51):385-391
DOI:10.4103/0973-1296.216337  PMID:29142388
Background: Orthosiphon stamineus (OS) Benth is a medicinal plant and native in Southeast Asia. Previous studies have shown that OS leaves possess antioxidant, cytotoxic, diuretic, antihypertensive, and uricosuric effects. These beneficial effects have been attributed to the presence of primary and secondary metabolites such as polyphenols, amino acids, and flavonoids. Objective: To develop and validate an high-performance liquid chromatography (HPLC)-diode array detector (DAD) method combined with solid-phase extraction that involves precolumn derivatization with O-phthaladehyde for simultaneous analysis of free amino acids in OS leaves extracts. Materials and Methods: OS leaves were extracted with water (OS-W), ethanol (OS-E), methanol (OS-M), 50% ethanol (OS-EW), and 50% methanol (OS-MW). The extracts were treated by C18 cartridge before derivatization, resulting in great improvement of separation by Zorbox Eclipse XDB-C18 column. Results: The HPLC–DAD method was successfully developed and validated for analyzing the contents of free amino acids in OS extracts. The results showed that l-aspartic acid with 0.93 ± 0.01 nmol/mg was the major free amino acid in OS-W extract. However, in OS-E, OS-M, OS-EW, and OS-MW, l-glutamic acid with 3.53 ± 0.16, 2.17 ± 0.10, 4.01 ± 0.12, and 2.49 ± 0.12 nmol/mg, respectively, was the major free amino acid. Subsequently, l-serine, which was detected in OS-W, OS-E, and OS-M, was the minor free amino acid with 0.33 ± 0.02, 0.12 ± 0.01, and 0.06 ± 0.01 nmol/mg, respectively. However, l-threonine with 0.26 ± 0.02 and 0.19 ± 0.08 nmol/mL in OS-EW and OS-MW, respectively, had the lowest concentration compared with other amino acid components. Conclusion: All validation parameters of the developed method indicate that the method is reliable and efficient to simultaneously determine the free amino acids content for routine analysis of OS extracts. Abbreviations used: HPLC-DAD: High-Performance Liquid Chromatography with Diode-Array Detection, OS: Orthosiphon stamineus, OS-W: Orthosiphon stamineus water extract, OS-E: Orthosiphon stamineus ethanol extract, OS-M: Orthosiphon stamineus methanol extract, OS-EW: Orthosiphon stamineus 50% ethanol extract, OS-MW: Orthosiphon stamineus 50% methanol extract, OPA: O-phthaladehyde, SPE: Solid Phase Extraction, UV: Ultraviolet, LOD: Limit of Detection, LOQ: Limit of Quantification, RSD: Relative Standard Deviation.
  3,778 308 3
Wound-healing activity of Zanthoxylum bungeanum maxim seed oil on experimentally burned rats
Xiao-Qiang Li, Rong Kang, Jun-Cheng Huo, Yan-Hua Xie, Si-Wang Wang, Wei Cao
July-September 2017, 13(51):363-371
DOI:10.4103/pm.pm_211_16  PMID:28839358
Background: The seed oil of Zanthoxylum bungeanum Maxim (ZBSO) is considered to be rich source of fatty acids, mainly oleic and linoleic acids, and has been used for the treatment of burns in Chinese medicine. Objective: We evaluated the healing efficacy of ZBSO and explored its possible mechanism on scalded rats. Materials and Methods: Sprague-Dawley rat models with deep second-degree burns were set up, and ZBSO (500 and 1000 μl/wound) was topically applied twice daily for 7 days and then once daily until wound healing. The therapeutic effects of ZBSO were evaluated by observing wound closure time, decrustation time, wound-healing ratio, and pathological changes. Collagen type-III, matrix metalloproteinase-2 (MMP-2), MMP-9, phospho-nuclear factor-κB (p-NF-κB) p65, inhibitor of NF-κB subunit α p-IκBα, and inhibitor of NF-κB subunit α (IκBα) expression were determined using Western blotting. Results: The ZBSO-treated group showed a higher wound-healing ratio and shorter decrustation and wound closure times than the untreated group. The topical application of ZBSO increased collagen synthesis as evidenced by an increase in hydroxyproline level and upregulated expression of collagen type-III on days 7, 14, and 21 posttreatment. A reduction in MMP-2 and MMP-9 expressions also confirmed the collagen formation efficacy of ZBSO. Furthermore, there was a significant increase in superoxide dismutase levels and a decrease in malondialdehyde levels in ZBSO-treated wounds. ZBSO also decreased tumor necrosis factor alpha, interleukin-1 (IL-1) β, and IL-6 levels in serum, upregulated IκBα, and downregulated p-NF-κB p65 and p-IκBα expression in vivo, indicating the anti-inflammatory action of ZBSO. Conclusion: ZBSO has significant potential to treat burn wounds by accelerating collagen synthesis and the anti-inflammatory cascade of the healing process. Abbreviations used: ECL: Enhanced chemiluminescence; ECM: Extracellular matrix; ELISA: Enzyme-linked immunosorbent assay; GC-MS: Gas chromatography-mass spectrometry; HRP: Horseradish peroxidase; HYP: Hydroxyproline; IκBα: Inhibitor of NF-κB subunit α; IL: Interleukin; MDA: Malondialdehyde; MMP: Matrix metalloproteinase-2; NF-κB: Nuclear factor-κB; SFE: Supercritical fluid extraction; SOD: Superoxide dismutase; SSD: Silver sulfadiazine; TCM: Traditional Chinese medicine; TNF: Tumor necrosis factor.
  3,850 228 5
Prim-O-glucosylcimifugin attenuates lipopolysaccharide- induced inflammatory response in RAW 264.7 macrophages
Jie Zhou, Yuan-Yuan Sun, Meng-Yao Sun, Wei-An Mao, Li Wang, Jian Zhang, Hong Zhang
July-September 2017, 13(51):378-384
DOI:10.4103/pm.pm_323_16  PMID:28839360
Background: Radix Saposhnikoviae (RS) exerts anti-inflammatory, analgesic, antipyretic, antioxidation effects and has been used in traditional Chinese medicine to treat common colds, headache, and rheumatoid arthritis. Prim-O-glucosylcimifugin (POG) is the highest content chromone and one of the major active constituents in RS. Objective: The study was aimed to explore the anti-inflammation effects of POG in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Materials and Methods: Cell viability was detected by Cell Counting Kit-8 assay. Production of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 was assessed by enzyme-linked immunosorbent assay. Real-time polymerase chain reaction and Western blot were performed to analyze mRNA and protein levels, respectively. Results: During the whole experiment, 15, 50, and 100 μg/mL of POG had no cytotoxicity on RAW 264.7 cells. POG dose-dependently inhibited the production of NO, TNF-α, IL-1β, and IL-6 that were induced by LPS. POG treatment downregulated the mRNA and protein expression inducible NO synthase (iNOS) and cyclooxygenase 2 (COX-2) in LPS-activated RAW 264.7 macrophages in a concentration-dependent manner. Furthermore, LPS-induced JAK2/STAT3 activation was prevented in RAW 264.7 macrophages by POG treatment. STAT3 overexpression significantly reversed the effects of POG on LPS-activated RAW 264.7 macrophages. Conclusion: These results demonstrate that POG exerts anti-inflammatory effects through the inhibition of iNOS and COX-2 expression by inhibiting the phosphorylation of JAK2/STAT3. Abbreviations used: LPS: Lipopolyssacharide; NO: Nitric oxide; TNF-α: Tumor necrosis factor-α; IL: Interleukin; RS: Radix Saposhnikoviae; POG: Prim-O-glucosylcimifugin; iNOS: Inducible NO synthase; COX2: Cyclooxygenase; FBS: Fetal bovine serum; DMSO: Dimethylsulfoxide; CCK-8: Cell Counting Kit; RIPA: Radio immunoprecipitation assay buffer; ECL: Enhanced chemiluminescence; SD: Standard deviation; ELISA: Enzyme-Linked immunosorbent assay. Dr. Hong Zhang,
  3,848 218 8
Anticancer potential of steviol in MCF-7 human breast cancer cells
Ena Gupta, Shweta Kaushik, Shalini Purwar, Ramesh Sharma, Anil K Balapure, Shanthy Sundaram
July-September 2017, 13(51):345-350
DOI:10.4103/pm.pm_29_17  PMID:28839355
Objective: This study aimed to investigate the cytotoxicity, apoptosis induction, and mechanism of action of steviol on human breast cancer cells (Michigan Cancer Foundation-7 [MCF-7]). Materials and Methods: Sulforhodamine-B assay was performed to analyze cytotoxic potential of Steviol whereas flow cytometer was used to analyze cell cycle, apoptosis, and reactive oxygen species generation. Results: Studying the viability of cells confirms the IC50of Steviol in MCF-7 cells which was 185 μM. The data obtained from fluorescence-activated cell sorter analysis reveal Steviol-mediated G2/M-phase arrest (P < 0.05) in addition to the presence of evident sub-G0/G1 peak (P < 0.05) in the MCF-7 cells, signifying the ongoing apoptosis. Conclusion: Thus, results suggest that induction of apoptosis in MCF-7 cells was due to dose-dependent effect of Steviol. Our first in vitro findings indicate Steviol as a promising candidate for the treatment of breast cancer. Abbreviations used: MCF-7: Michigan Cancer Foundation-7; SRB: Sulforhodamine-B assay; FACS: Fluorescence-activated cell sorter; ROS: Reactive oxygen species; DNA: Deoxyribonucleic acid.
  3,737 182 8
Quantification analysis and In vitro anti-inflammatory effects of 20-hydroxyecdysone, momordin ic, and oleanolic acid from the fructus of Kochia scoparia
Sae-Rom Yoo, Soo-Jin Jeong, Na-Ri Lee, Hyeun-Kyoo Shin, Chang-Seob Seo
July-September 2017, 13(51):339-344
DOI:10.4103/0973-1296.211023  PMID:28839354
Background: The fructus of Kochia scoparia Schrader (Chenopodiaceae) is a traditional herbal medicine that has been used for treating gonorrhea and dermatitis. Objective: We investigated the anti-inflammatory activities of three marker compounds, including 20-hydroxyecdysone, momordin Ic, and oleanolic acid, from the fructus of K. scoparia. Materials and Methods: The simultaneous analysis of three components was performed using high-performance liquid chromatography and high-performance thin-layer chromatography. We evaluated the anti-inflammatory effects of the nine marker compounds by determining their anti-inflammatory activities in the murine macrophage cell line RAW 264.7. Results: Among three marker compounds, momordin Ic, but not 20-hydroxyecdysone and oleanolic acid, had inhibitory effects on the production of inflammatory cytokines tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in LPS-treated RAW264.7 macrophages. The effects of three marker compounds on prostaglandin E2(PGE2) were also evaluated. All three compounds significantly reduced PGE2production in LPS-treated cells. Conclusions: We suggest that momordin Ic is the most potent phytochemical of the fructus of K. scoparia as an anti-inflammatory agent. Abbreviations used: HPLC: High-performance liquid chromatography; TNF-α: Tumor necrosis factor alpha; IL-6: Interleukin-6; PGE2: Pro-inflammatory mediator prostaglandin E2; LPS: Lipopolysaccharide.
  3,745 129 5
Optimization of surfactant-mediated, ultrasonic-assisted extraction of antioxidant polyphenols from rattan tea (Ampelopsis grossedentata) using response surface methodology
Feng Li, Aun Raza, Yan-Wei Wang, Xiu-Quan Xu, Guan-Hua Chen
July-September 2017, 13(51):446-453
DOI:10.4103/pm.pm_159_16  PMID:28839370
Background: Rattan tea is a medicinal plant that has been used for many years for the treatment of inflammation, fatty liver, tumor, diabetes, and hyperlipidemia. Objective: A green and novel approach based on surfactant-mediated, ultrasonic-assisted extraction (SM-UAE) was developed for the extraction of antioxidant polyphenols from Rattan tea. A nonionic surfactant Tween-80 was selected as extraction solvent. The antioxidant activity was measured by total phenolic content (TPC) and ferric-reducing/antioxidant capacity (FRAC) assay. Materials and Methods: Optimization of extraction parameters including concentration of solvent, ultrasonic time, and temperature were investigated by response surface methodology. The antioxidant activity was measured by TPC and FRAC assay. Results: The optimal extraction conditions were determined as 6.8% (v/v) of aqueous Tween-80, ultrasonic temperature of 54°C, and ultrasonic time of 28.8 min. Under these conditions, the highest TPC value of 360.4 mg gallic acid equivalent per gram of dry weight material (GAE/g DW) was recorded. Moreover, 6.8% (v/v) of aqueous Tween-80, ultrasonic temperature of 54.5°C, and ultrasonic time of 28.4 min were determined for the highest FRAC value of 478.2 μmol of Fe2+/g of weight material (μmol Fe2+/g DW). Compared with other methods, the TPC and FRAC values of 313.5 mg GAE/g DW and 389.6 μmol Fe2+/g DW were obtained by heat reflux extraction using ethanol as solvent, respectively, and 343.2 mg GAE/g DW and 450.1 μmol Fe2+/g DW were obtained by UAE using ethanol as solvent, respectively. Conclusion: The application of SM-UAE markedly decreased extraction time or extraction cost and improved the extraction efficiency, compared with the other methods. Abbreviations used: SM-UAE: Surfactant-mediated ultrasonic-assisted extraction; TPC: total phenolic content; FRAC: Ferric reducing antioxidant capacity; RSM: Response surface methodology; BBD: Box-Behnken design.
  3,618 205 8
Quassinoids from the root of Eurycoma longifolia and their antiproliferative activity on human cancer cell lines
Nguyen Huu Tung, Takuhiro Uto, Nguyen Thanh Hai, Gang Li, Yukihiro Shoyama
July-September 2017, 13(51):459-462
DOI:10.4103/pm.pm_353_16  PMID:28839372
Background: The roots of Eurycoma longifolia Jack have traditionally been used as an aphrodisiac tonic besides the other remedies for boils, fever, bleeding gums, and wound ulcer. Recently, the antiproliferative activity of E. longifolia has been reported and remained attractive to natural chemists. Objective: The objective of this study was to study on antiproliferative compounds from the root of E. longifolia. Materials and Methods: Column chromatography was used to separate individual compounds, spectroscopic data including nuclear magnetic resonances and mass spectrometry were analyzed to determine the chemical structure of the isolates and for biological testing, antiproliferative activity of compounds was tested on seven human cancer cell lines (KATO III, HCT-15, Colo205, HepG2, PC-3, Jurkat, HL-60) by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Results: Nine quassinoids including a new C19longilactone-type quassinoid glycoside were characterized from the roots of the title plant. Among them, the major quassinoid eurycomanone exhibited selectively potential antiproliferative activities on the leukemia cell lines (HL-60 and Jurkat) and had very low toxic effects on normal skin fibroblast cell line (NB1RGB). Conclusion: The current study reveals one new quassinoid glycoside and the potential active component (eurycomanone) from E. longifolia for the leukemia treatment. Abbreviations used: COSY: Correlation spectroscopy; HMBC: Heteronuclear multiple-bond correlation; HMQC: Heteronuclear multiple quantum correlation; NMR: Nuclear magnetic resonance; NOESY: Nuclear Overhauser enhancement spectroscopy; TLC: Thin layer chromatography.
  3,568 169 6
Correlation between polyphenol oxidase (PPO) activity and total phenolic contents in Crocus sativus L. corms during dormancy and sprouting stages
Nardana Esmaeili, Hassan Ebrahimzadeh, Khosrou Abdi
July-September 2017, 13(51):519-524
DOI:10.4103/0973-1296.216333  PMID:29142408
Purification and characterization of polyphenol oxidase (PPO) enzyme and determination of total phenolic contents during dormancy and sprouting stages in Crocus sativus corms were performed. PPO enzyme was purified by ammonium sulfate precipitation and ion-exchange chromatography using DEAE-Sephadex A25 and two isoenzymes were obtained on the SDS-PAGE, which corresponded to molecular weights of 70 and 54 kDa. The Km values of the enzyme were 4.87 and 2.12 mM for l-DOPA in dormancy and waking stages, respectively. Also, enzyme showed higher Vmax values of 0.026 (ΔOD.min-1) in dormancy compared with the value of 0.019 (ΔOD.min-1) in waking corms. Results showed an inverse correlation between phenolic contents and PPO activity. Accordingly, it can be concluded that as plant progressed through sprouting stage, in contrast to polyphenol oxidase activity, there was a significant increase in total amount of phenolic compounds, as determined by Folin-Ciocalteu method and water and aqueous ethanol extractions. Abbreviations used: PPO: Polyphenol Oxidase, DEAE-Sephadex: Diethylaminoethyl Sephadex, SDS-PAGE: Sodium Dodecyl Sulfate- Polyacrylamide Gel Electrophoresis, DOPA: Dihydroxyphenylalanine, PEG: Polyethylene Glycol
  3,464 229 5
Cinnamaldehyde, cinnamic acid, and cinnamyl alcohol, the bioactives of Cinnamomum cassia exhibit HDAC8 inhibitory activity: An In vitro and In silico study
Mangesh Patil, Amit S Choudhari, Savita Pandita, Md Ataul Islam, Prerna Raina, Ruchika Kaul-Ghanekar
July-September 2017, 13(51):645-651
DOI:10.4103/pm.pm_389_16  PMID:29142427
Background: The altered expression of histone deacetylase family member 8 (HDAC8) has been found to be linked with various cancers, thereby making its selective inhibition a potential strategy in cancer therapy. Recently, plant secondary metabolites, particularly phenolic compounds, have been shown to possess HDAC inhibitory activity. Objective: In the present work, we have evaluated the potential of cinnamaldehyde (CAL), cinnamic acid (CA), and cinnamyl alcohol (CALC) (bioactives of Cinnamomum) as well as aqueous cinnamon extract (ACE), to inhibit HDAC8 activity in vitro and in silico. Materials and Methods: HDAC8 inhibitory activity of ACE and cinnamon bioactives was determined in vitro using HDAC8 inhibitor screening kit. Trichostatin A (TSA), a well-known anti-cancer agent and HDAC inhibitor, was used as a positive control. In silico studies included molecular descriptor Analysis molecular docking absorption, distribution, metabolism, excretion, and toxicity prediction, density function theory calculation and synthetic accessibility program. Results: Pharmacoinformatics studies implicated that ACE and its Bioactives (CAL, CA, and CALC) exhibited comparable activity with that of TSA. The highest occupied molecular orbitals and lowest unoccupied molecular orbitals along with binding energy of cinnamon bioactives were comparable with that of TSA. Molecular docking results suggested that all the ligands maintained two hydrogen bond interactions within the active site of HDAC8. Finally, the synthetic accessibility values showed that cinnamon bioactives were easy to synthesize compared to TSA. Conclusion: It was evident from both the experimental and computational data that cinnamon bioactives exhibited significant HDAC8 inhibitory activity, thereby suggesting their potential therapeutic implications against cancer. Abbreviations used: ACE: Aqueous Cinnamon Extract; DFT: Density Function Theory; CAL: Cinnamaldehyde; CA: Cinnamic Acid; CALC: Cinnamyl Alcohol; MW: Molecular Weight; ROTBs: Rotatable Bonds; ROF: Lipinski's Rule of Five; TSA: Trichostatin A; PDB: Protein Data Bank; RMSD: Root Mean Square Deviation; HBA: Hydrogen Bond Acceptor; HBD: Hydrogen Bond Donor; ADMET: Absorption, Distribution, Metabolism, Excretion and Toxicity; FO: Frontier Orbital; HOMOs: Highest Occupied Molecular Orbitals; LUMOs: Lowest Unoccupied Molecular Orbitals; BE: Binding Energy.
  3,386 220 6
Bioactive turmerosaccharides from Curcuma longa Extract (NR-INF-02): Potential ameliorating effect on osteoarthritis pain
Bharathi Bethapudi, Sasikumar Murugan, Ramanaiah Illuri, Deepak Mundkinajeddu, Chandrasekaran Chinampudur Velusami
July-September 2017, 13(51):623-627
DOI:10.4103/pm.pm_465_16  PMID:29142423
Background: Curcuma longa has long history of medicinal use in Ayurveda. A unique product NR-INF-02 was prepared from C. longa that was standardized to contain turmerosaccharides. Objective: The present study investigated the effect of turmerosaccharides rich fraction of NR-INF-02 on monosodium iodoacetate (MIA)-induced OA pain animal model that mimics human OA. Further, the analgesic effect of turmerosaccharides rich fraction was compared to turmerosaccharides less fraction of NR-INF-02. Materials and Methods: OA pain was chemically induced by intra-articular administration of single dose of 25 μ l of 0.9% saline containing 0.3 mg MIA into the right knee of male albino Wistar rat. Turmerosaccharides rich fraction and turmerosaccharides less fraction (at 22.5, 45 and 90 mg/kg rat body weight dose levels) were administered as a single dose orally on day 5 of post-MIA injection. OA pain was measured using hind limb weight-bearing ability at 1, 3, 6, and 24 h post-test substance administration on day 5. Results: Oral administration of turmerosaccharides rich fraction and turmerosaccharides less fraction (at 45 and 90 mg/kg) although significantly decreased the OA pain at all the intervals, the effect of turmerosaccharides rich fraction (57%) on OA pain was superior to turmerosaccharides less fraction (35%). Conclusion: Bioactive turmerosaccharides from C. longa extract contribute to the observed anti-arthritic effect in rats. Abbreviations used: MIA: Monosodium iodoacetate; i.ar: Intra-articular; OA: Osteoarthritis; TRF: Turmerosaccharides rich fraction; TLF: Turmerosaccharides less fraction; PGE2: Prostaglandin E2; ROS: Reactive oxygen species.
  3,304 254 6
Omega-3 fish oil reduces mortality due to severe sepsis with acute gastrointestinal injury grade III
Huaisheng Chen, Wei Wang, Chengying Hong, Ming Zhang, Yingcai Hong, Su Wang, Huadong Zhang
July-September 2017, 13(51):407-412
DOI:10.4103/pm.pm_418_16  PMID:28839364
Background: Sepsis plays an important role in acute gastrointestinal injury (AGI). Our research was designed to determine the effects of omega-3 fish oil (FO) in patients suffering from severe sepsis combined with AGI III, and the ability of FO to modulate immune function. Methods: Seventy-eight patients diagnosed with severe sepsis with AGI III and a need for mechanical ventilation were randomized to two groups. In the FO group, 50 g of long chain fatty acid soybean oil (n = 6) and 10 g of FO (n = 3) were administered as total parenteral nutrition (TPN). The control group was treated with 50 g of long chain fatty acid soybean oil without addition of FO to TPN. Results: At baseline, there were no significant differences between the two groups. The 60-day mortality was lower in the FO group. Multiple factor logistic regression analysis revealed that intra-abdominal pressure (IAP) and abdominal infection were correlated with the FO intervention. The patients with abdominal infection demonstrated a lower mortality rate, fewer CD3 T lymphocytes, and fewer helper/inducer T lymphocytes in the FO group compared with the control group. After 7 days, the Marshall Score was lower in the FO group than in the control group. Conclusion: FO has positive effects in terms of improving the long-term prognosis of patients with severe sepsis with AGI III. Patients with a high IAP and abdominal infection might experience greater benefit from FO. This effect might be due, in part, to immunomodulation. Abbreviations used: AGI: Acute gastrointestinal injury; FO: Fish oil; TPN: Total parenteral nutrition; IAP: Intra-abdominal pressure; ICU: Intensive Care Unit; MODS: Multiple organ dysfunction syndrome; TLR4: Toll-like receptor 4; DNR: Do Not Resuscitate; WGAP: Working Group of Abdominal Problem; EN: Enteral nutrition; BP: Low blood pressure; CRI: Catheter-related infection; PBS: Phosphate-buffered saline; ELFA: Enzyme-linked fluorescent assay; SD: Standard deviation; PUFAs: Polyunsaturated fatty acids; EPA: Eicosapentenoic acid; DHA: Docosahexaenoic acid.
  3,302 173 4
Reversible testicular toxicity of piperine on male albino rats
Gopichand Chinta, Mohane Selvaraj Coumar, Latha Periyasamy
July-September 2017, 13(51):525-532
DOI:10.4103/pm.pm_405_16  PMID:29142409
Background: Piperine was widely used in traditional medicine for inducing sterility and abortion. Objective: To evaluate the effect of the piperine on testis of male albino rats. Materials and Methods: Adult male rats were divided into four groups (n = 12). Group I (control): Rats were given vehicle p.o. i.e. 0.5% carboxymethyl cellulose in normal saline daily for 60 days, Group II (ED): Rats received piperine at a dose of 10 mg/kg body weight (b.w.) daily, Group III (E4D): Rats received piperine at a dose of 10 mg/kg b.w. on every 4th day, Group IV (E7D): Rats received piperine at a dose of 10 mg/kg b.w. on every 7th day. Half of the animals from each group were sacrificed after the treatment period (60 days), and the remaining were kept for drug-free withdrawal period (60 days) and then sacrificed. Results: Piperine significantly decreased the reproductive organ weights in groups ED and E4D. Piperine induced hormonal imbalance by altering the serum levels of follicle-stimulating hormone, luteinizing hormone, sex hormone binding globulin, serum, and testicular testosterone in groups ED and E4D. Furthermore, piperine decreased the activity of germ cell markers and Leydig cellular steroidogenic enzymes in the groups ED and E4D after 60 days. All the above-altered values returned to normal levels after withdrawal period. Histopathological findings also supported the above findings. Conclusion: From the above data, it can be concluded that piperine could be a good lead molecule for the development of reversible oral male contraceptive.
  3,250 163 3
Topical delivery of Withania somnifera crude extracts in niosomes and solid lipid nanoparticles
Tawona N Chinembiri, Minja Gerber, Lissinda H du Plessis, Jan L du Preez, Josias H Hamman, Jeanetta du Plessis
July-September 2017, 13(51):663-671
DOI:10.4103/pm.pm_489_16  PMID:29142430
Background: Withania somnifera is a medicinal plant native to India and is known to have anticancer properties. It has been investigated for its anti-melanoma properties, and since melanoma presents on the skin, it is prudent to probe the use of W. somnifera in topical formulations. To enhance topical drug delivery and to allow for controlled release, the use of niosomes and solid lipid nanoparticles (SLNs) as delivery vesicles were explored. Objective: The objective of this study is to determine the stability and topical delivery of W. somnifera crude extracts encapsulated in niosomes and SLNs. Materials and Methods: Water, ethanol, and 50% ethanol crude extracts of W. somnifera were prepared using 24 h soxhlet extraction which were each encapsulated in niosomes and SLNs. Franz cell diffusion studies were conducted with the encapsulated extracts to determine the release and skin penetration of the phytomolecules, withaferin A, and withanolide A. Results: The niosome and SLN formulations had average sizes ranging from 165.9 ± 9.4 to 304.6 ± 52.4 nm with the 50% ethanol extract formulations having the largest size. A small particle size seemed to have correlated with a low encapsulation efficiency (EE) of withaferin A, but a high EE of withanolide A. There was a significant difference (P < 0.05) between the amount of withaferin A and withanolide A that were released from each of the formulations, but only the SLN formulations managed to deliver withaferin A to the stratum corneum-epidermis and epidermis-dermis layers of the skin. Conclusion: SLNs and niosomes were able to encapsulate crude extracts of W. somnifera and release the marker compounds, withaferin A, and withanolide A, for delivery to certain layers in the skin. Abbreviations used: API: Active pharmaceutical ingredient, ANOVA: Analysis of variance, ED: Epidermis-dermis, HPLC: High-performance liquid chromatography, HLB: Hydrophilic-lipophilic balance, NMR: Nuclear magnetic resonance spectroscopy, PDI: Polydispersity index, SLN: Solid lipid nanoparticle, SD: Standard deviation, SCE: Stratum corneum-epidermis, TEM: Transmission electron microscopy.
  3,053 288 2
A novel strategy for bitter taste masking of gankeshuangqing dispersible tablets based on particle coating technology
Xue Han, Ding-Kun Zhang, Fang Zhang, Jun-Zhi Lin, Hong Jiang, Yang Lan, Xi Xiong, Li Han, Ming Yang, Chao-Mei Fu
July-September 2017, 13(51):400-406
DOI:10.4103/pm.pm_240_16  PMID:28839363
Background: Currently, acute upper respiratory tract infections (AURTIs) are increasingly becoming a significant health burden. Gankeshuangqing dispersible tablets (GKSQDT) which have a good effect on treating AURTIs. GKSQDT is composed of baicalin and andrographolide. However, its severe bitterness limits application of patients. Due to the addition of plentiful accessories, common masking methods are unsuitable for GKSQDT. It is thus necessary to develop a new masking method. Materials and Methods: The Previous study showed that baicalin was less bitter than andrographolide. Thus, particle coating technology was adapted to prepare composite particles that baicalin coated on the surface of andrographolide to decrease bitterness. Initially, particle size of baicalin and coating time of composite was investigated to prepare composite. Then, scanning electron microscopy, wettability, and infrared (IR) spectrogram were used to characterize the microstructure of composite. Furthermore, electronic tongue test, animal preference experiment, and human sensory test were applied to evaluate the masking effect. Results: To produce composite, baicalin should be ground in vibromill for 6 min. Then, andrographolide fine powder was added to grind together for 6 min. Contact angle of composite was smaller than mixture, and more similar to baicalin. Other physical characterization including microstructure, wettability, and IR also suggested that andrographolide was successfully coated by baicalin superfine. Furthermore, taste-masking test indicated taste-masked tablets was less bitter than original tablets. Conclusion: The study indicated that particle coating technology can be used for taste masking of GKSQDT without adding other substance. Moreover, it provides a new strategy of taste masking for national medicine. Abbreviations used: AURTIs: Acute Upper Respiratory Tract Infections; GSQDT: Gankeshuangqing Dispersible Tablets; IR: Infrared Spectrogram; LHPC: Low-substituted Hydroxypropyl Cellulose; CAs: Contact Angles; FTIR: Fourier Transform Infrared Spectra.
  3,160 171 2
Are polyunsaturated fatty acid metabolites, the protective effect of 4-hydroxytyrosol on human red blood cell membranes and oxidative damage (4-hydroxyalkenals) compatible in hypertriglyceridemic patients?
Giuseppe Gallo, Rosalinda Bruno, Adele Taranto, Guglielmo Martino
July-September 2017, 13(51):561-566
DOI:10.4103/pm.pm_483_15  PMID:29142415
Background: Increased levels of malondialdehyde (MDA) and 4-hydroxynonenal (HNE) are demonstrated in plasma of uremic patients. A study showed that the comparison of erythrocytes of healthy and diseased patients (obese, hypertensive, and Type 2 diabetics) with age is associated to a disturbed oxidant/antioxidant balance when obesity is associated with hypertension. 4-hydroxytyrosol is shown to significantly protect red blood cells (RBCs) from oxidative damage (4-HNE). In literature, there are partial discussions on the role of lipids and their oxidation products. The products of degradation of membrane proteins are observed as self-consisting products without interrelations with membrane lipids. Objective: The aim of this study is to evaluate the role of polyunsaturated fatty acid (PUFA) metabolites on oxidative damage (4-hydroxy-alkenals) in RBCs of hypertriglyceridemic patients after membrane treatment with 4-hydroxytyrosol. Materials and Methods: The authors optimize the isolation of RBC ghosts and spectrophotometric method to measure free 4-hydroxyalkenals in human RBC membranes and investigated the effect on oxidative damage in human erythrocyte membranes and in vitro 4-hydroxytyrosol treatment to evaluate the membrane lipids reducible by this phenol. Results: Plasma triglyceride levels in patients are clearly higher than in controls. Moreover, total membrane proteins data are similar to previous described. The normalized alkenals levels are significantly enhanced in hyperlipemic patients in comparison to normoglyceridemic controls. After the 4-hydroxytyrosol action, lipid metabolites substantially decrease. The ratio of oxidized lipids (MDA + HNE) and membrane proteins data are similar to previously described ones. Conclusion: According to experimental data, the accumulation of the alkenals in RBC membrane could be produced either by partial PUFA oxidation contained in glycerides and plasma glycerides and by glycerides into plasma membrane recycled RBC. Abbreviations used: RBC: Red blood cell; MDA: Malondialdehyde; HNE\HAE: 4-hydroxyalkenals; LPO: Lipid peroxidation; ROS: Reactive oxygen species; ORAC: Oxygen Radical Absorbance Capacity.
  3,204 93 4
Antipsoriatic and anti-inflammatory studies of Berberis aristata extract loaded nanovesicular gels
Nimisha , Dilshad Ali Rizvi, Zeeshan Fatima, Neema , Chanchal Deep Kaur
July-September 2017, 13(51):587-594
DOI:10.4103/pm.pm_210_17  PMID:29142419
Objective: Novel nanovesicular gel of Berberis aristata extract was developed and evaluated for its anti-inflammatory and antipsoriatic activity. Materials and Methods: Transferosomes were prepared using soya phosphatidylcholine and edge activators (Tween 80, Span 80, and sodium deoxycholate) by a modified lipid film hydration technique using rotary evaporator and evaluated for various parameters. The quantification and standardization of extract have been carried out using its alkaloid content as berberine as biomarker. Topical application of imiquimod (IMQ) (immune modifier) on the shaved back of mice developed psoriasis-like inflammation followed by histopathological study of inflamed skin. Results: The size of transferosomes was in the range of 265–345 nm whereas polydispersity index ranges from 0.10 to 0.63, and for zeta potential, it was from −19.3 to −43.3 mV. Transferosomes were further added to Carbopol 934P for gel formation and subsequently evaluated for their physicochemical properties. Their efficacy against inflammation, IMQ-induced psoriasis, and skin sensitivity was compared with conventional formulation (commercial formulation-Angle Gloss, Phytolab Pvt. Ltd.). Percent inhibition of edema by transferosomal gel (55.76%) was more as compared to conventional gel of extract (33.5%) found out by Carrageenan-induced paw edema method. Primary irritation index was found to be <0.4 inferring its safe use for topical formulation. Conclusion: Histopathological report showed that, in psoriasis-induced animal treated with topical application of extract loaded transferosomal gel showed a marked reduction in thickness of epidermis, length of rete ridges as compared to conventional gel formulation. It can be inferred that B. aristata extract loaded transferosomal gel can function as potential anti-inflammatory and antipsoriatic formulation. Abbreviations used: SPC: Soyaphosphatidylcholine, PDI: Polydispersity index, IMQ: Imiquimod, EA: Edge activator, BE: Berberine, TEM: Transmission electron microscopy, PBS: Phosphate buffered saline, H and E: Hematoxylin and eosin, ZP: Zeta potential, EE: Entrapment efficiency.
  3,044 239 3
Hepatoprotective flavonoids in Opuntia ficus-indica fruits by reducing oxidative stress in primary rat hepatocytes
Jung Wha Kim, Tae Bum Kim, Hyun Woo Kim, Sang Wook Park, Hong Pyo Kim, Sang Hyun Sung
July-September 2017, 13(51):472-476
DOI:10.4103/pm.pm_232_16  PMID:28839374
Background: Liver disorder was associated with alcohol consumption caused by hepatic cellular damages. Opuntia ficus-indica fruit extracts (OFIEs), which contain betalain pigments and polyphenols including flavonoids, have been introduced as reducing hangover symptoms and liver protective activity. Objective: To evaluate hepatoprotective activity of OFIEs and isolated compounds by high-speed countercurrent chromatography (HSCCC). Materials and Methods: The extract of O. ficus-indica fruits was fractionated into methylene chloride and n-butanol. The n-butanol fraction was isolated by HSCCC separation (methylene chloride-methanol-n-butanol-water, 5:4:3:5, v/v/v/v). The hepatoprotective activity of OFIEs and isolated compounds was evaluated on rat primary hepatocytes against ethanol-induced toxicity. Antioxidative parameters such as glutathione reductase and glutathione peroxidase (GSH-Px) enzymes and the GSH content were measured. Results: Two flavonoids, quercetin 3-O-methyl ester (1) and (+)-taxifolin, and two flavonoid glycosides, isorhamnetin 3-O-β-d-glucoside (3) and narcissin (4), were isolated from the n-butanol fraction by HSCCC separation. Among them, compound 2 significantly protected rat primary hepatocytes against ethanol exposure by preserving antioxidative properties of GR and GSH-Px. Conclusions: OFIEs and (+)-taxifolin were suggested to reduce hepatic damage by alcoholic oxidative stress.
  3,134 144 5
Using molecular docking analysis to discovery Dregea sinensis Hemsl. potential mechanism of anticancer, antidepression, and immunoregulation
Xiujie Liu, Yu Shi, Yulin Deng, Rongji Dai
July-September 2017, 13(51):358-362
DOI:10.4103/pm.pm_384_16  PMID:28839357
Background: Dregea sinensis Hemsl. plant of the genus Dregea volubilis (Asclepiadaceae), plays a vital role in anticancer, antidepression, and immunoregulation. Steroidal glycosides are the main constituents of this herb, which were significant biological active ingredients. Objective: The objective of this study is to recognize the mechanism of anticancer, antidepression, and immunoregulation of D. sinensis Hemsl. Materials and Methods: Seventy-two steroidal glycosides of D. sinensis Hemsl. were evaluated on the docking behavior of tumor-associated proteins (PI3K, Akt, mTOR), depression-related proteins (MAO-A, MAO-B) and immune-related proteins (tumor necrosis factor-α [TNF-α], tumor necrosis factor receptor 2 [TNFR2], interleukin-2Rα [IL-2Rα]) using Discovery Studio version 3.1 (Accelrys, San Diego, USA). Results: The molecular docking analysis revealed that mostly steroidal glycosides of D. sinensis Hemsl. exhibited powerful interaction with the depression-related protein (MAO-A) and the immune-related proteins (TNFR2, IL-2Rα). Some ligands exhibited high binding energy for the tumor-associated proteins (PI3K, Akt, mTOR) and the immune-related protein (TNF-α), but MAO-B showed none interaction with the ligands. Conclusion: This study has paved better understanding of steroidal glycosides from D. sinensis Hemsl. as potential constituents to the prevention of associated cancer, depression and disorders of immunoregulation. Abbreviations used: PI3K: Phosphatidyl inositol 3-kinase; Akt: Protein kinase B; mTOR: Mammalian target of rapamycin; MAO-A: Monoamine oxidase A; MAO-B: Monoamine oxidase B; TNF-α: Tumor necrosis factor α; TNFR2: Tumor necrosis factor receptor 2; IL-2Rα: The alpha subunit (CD25) of the interleukin-2 receptor; DS: Discovery Studio; PDB: Protein Database Bank; 3D: three-dimensional.
  3,111 165 3
Phytochemical compositions and In vitro assessments of antioxidant and antidiabetic potentials of fractions from Ehretia cymosa Thonn
Akintayo Ogundajo, Anofi Tom Ashafa
July-September 2017, 13(51):470-480
DOI:10.4103/pm.pm_118_17  PMID:29142401
Background: Ehretia cymosa Thonn. is a popular medicinal plant used in different parts of West Africa for the treatment of various ailments including diabetes mellitus. Objective: The current study investigates bioactive constituents and in vitro antioxidant and antidiabetic potentials of fractions from extract of E. cymosa. Materials and Methods: Phytochemical investigation and antioxidant assays were carried out using standard procedures. Antidiabetic potential was assessed by evaluating the inhibitory effects of the fractions on the activities of α -amylase and α -glucosidase, while bioactive constituent's identification was carried out using gas chromatography-mass spectrometric (GC-MS) analysis. Results: The phytochemistry tests of the fractions revealed the presence of tannins, phenols, flavonoids, steroids, terpene, alkaloid, and cardiac glycosides. Methanol fraction shows higher phenolic (27.44 mg gallic acid/g) and flavonoid (235.31 mg quercetin/g) contents, while ethyl acetate fraction revealed higher proanthocyanidins (28.31 mg catechin/g). Methanol fraction displayed higher (P < 0.05) 1,1-diphenyl-2-picryl-hydrazyl (0.47 mg/mL), 2,2-azino-bis (3-ethylbenzothiazoline)-6-sulfonic acid (0.49 mg/mL), and hydroxyl radical (0.55 mg/mL) scavenging activities, while ethyl acetate exhibited strong metal chelating (0.61 mg/mL) and superoxide anion (1.68 mg/mL) scavenging activity. Methanol and ethyl acetate fractions displayed higher inhibition (P < 0.05) against α -glucosidase (0.60 mg/mL) and α -amylase (2.11 mg/mL), respectively. Methanol fraction also inhibited α -amylase and α -glucosidase in competitive and noncompetitive modes, respectively. The GC-MS chromatogram of the methanol fraction revealed 24 compounds, which include phytol (1.78%), stearic acid (1.02%), and 2-hexadecyloxirane (34.18%), which are known antidiabetic and antioxidant agents. Conclusion: The results indicate E. cymosa leaves as source of active phytochemicals with therapeutic potentials in the management of diabetes. Abbreviations used: ABTS: 2,2- Azino-bis (3-ethylbenzothiazoline)-6-sulfonic acid, DPPH: 1,1-diphenyl-2-picryl-hydrazyl, PMS: Phenazine methosulfate, NBT: Nitroblue tetrazolium, NADH: Nicotinamide adenine dinucleotide, TCA: Trichloroacetic acid, TBA: Thiobarbituric acid, DNS: Dinitrosalicylic acid.
  2,995 197 -
Exploring the cytotoxic potential of triterpenoids-enriched fraction of Bacopa monnieri by implementing In vitro, In vivo, and In silico approaches
Md. Nasar Mallick, Washim Khan, Rabea Parveen, Sayeed Ahmad, Sadaf , Mohammad Zeeshan Najm, Istaq Ahmad, Syed Akhtar Husain
July-September 2017, 13(51):595-606
DOI:10.4103/pm.pm_397_16  PMID:29142420
Background: Bacopa monnieri (BM) is a herbaceous plant traditionally used from time immemorial in Ayurvedic and folklore medicines. We hypothesized that the extract of the whole plant might contain numerous molecules with having antitumor activities that could be very effective in killing of human cancer cells. Objectives: This work investigated anticancer activity of bioactive fraction of BM. Materials and Methods: The hydroalcoholic extract of BM was fractionated with different solvent, namely, hexane, dichloromethane (DCM), acetone, methanol, and water. The in vitro anticancer activity was performed against various Human Cancer Cell lines, namely, Colon (HT29, Colo320, and Caco2), Lung (A549), Cervix (HeLa, SiHa), and Breast (MCF-7, MDAMB-231). Further, DCM fraction was evaluated in vivo for anticancer activity against Ehrlich ascites carcinoma (EAC) tumor-bearing mice since it showed the best cytotoxicity at 72 h (IC5041.0–60.0 μg/mL). The metabolic fingerprinting of these extract were carried out using high-performance thin-layer chromatography along with quantification of bacoside A, bacoside B, cucurbitacin B, cucurbitacin E, and bittulinic acid. Results: Oral administration of DCM fraction at a dose of 40 mg/kg rendered prominent reduction of tumor regression parameters such as tumor weight, packed cell volume, tumor volume and viable tumor cell count as compared to the untreated mice of the EAC control group. The anticancer activity of DCM fraction may be due to the presence of large amount of bacoside A, B and cucurbitacins. The molecular docking studies of major metabolites with targeted proteins predicted the anticancer activity of DCM fraction which was in support of in vivo activity. Conclusion: The in vitro, in vivo, analytical and in silico studies on DCM fraction of Bacopa monieri has proved its great potential for development of anticancer phytopharmaceuticals. Abbreviations used: DBM: DCM fraction of Bacopa monnieri; DCM: Dichloromethane; EAC: Ehrlich ascites carcinoma; HCT: Hematocrit; HGB: Hemoglobin; HPTLC: High performance thin layer chromatography; ICH: International council for Harmonisation; LOD: Limit of detection; LOQ: Limit of quantification; LYM: Lymphocytes; MCH: Mean corpuscular hemoglobin; MCHC: Mean corpuscular haemoglobin concentration (MCHC); MCV: Mean corpuscular volume; MTT: 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; PLT: Platelet; RBC: Red blood cell; RDW: Red blood cell distribution width; RSD: Relative standard deviation; WBC: White blood cells.
  3,006 185 5
Mechanism of fructus aurantii flavonoids promoting gastrointestinal motility: From organic and inorganic endogenous substances combination point of view
Shuai Wang, Yong-Rui Bao, Tian-Jiao Li, Ting Yu, Xin Chang, Guan-Lin Yang, Xian-Sheng Meng
July-September 2017, 13(51):372-377
DOI:10.4103/pm.pm_179_16  PMID:28839359
Background: Fructus Aurantii (FA) derived from the dried, and unripe fruit of Citrus aurantium L. is one of the commonly used traditional Chinese medicines to treat gastrointestinal motility dysfunction diseases. According to the literature research, FA flavonoids (FAF) are important active ingredients of FA promoting gastrointestinal motility, but the exact material basis and mechanism of action are still not very clear. Objective: This experiment was designed to illustrate the material basis of FAF promoting gastrointestinal motility and explore the mechanism of action from an organic and inorganic combination point of view. Materials and Methods: In this experiment, high-performance liquid chromatography (HPLC) method was used to analyze the composition and content of FAF. Based on the prominent prokinetic effect of FAF on mice, the mechanism of action was speculated through a combination of HPLC coupled with quadrupole time-of-flight mass spectrometry (HPLC-QTOF-MS) and inductively coupled plasma mass spectrometry (ICP-MS). Results: With the method of HPLC, ten dominating components of FAF including neoeriocitrin, narirutin, rhoifolin, naringin, hesperidin, neohesperidin, neoponcirin, naringenin, hesperetin, and nobiletin accounting for more than 86% of FAF were identified. Combined HPLC-QTOF-MS with ICP-MS, the endogenous substances with difference in the blood of mice were analyzed, in which 4-dimethylallyltryptophan, corticosterone, phytosphingosine, sphinganine, LysoPC (20:4(5Z, 8Z, 11Z, 14Z)), LysoPC(18:2 (9Z, 12Z)), and Ca2+, Mg2+, Zn2+ metal ions had significant changes, involving tryptophan metabolism, corticosterone metabolism, sphingolipid metabolism, and other pathways. Conclusion: The results preliminarily elaborated the mechanism of FAF promoting gastrointestinal motility from an organic and inorganic point of view, which provide valuable information for researching and developing new multi-component Chinese medicine curing gastrointestinal underpower associated diseases. Abbreviations used: FA: Fructus Aurantii; FAF: Fructus Aurantii flavonoids; HPLC: High performance liquid chromatography; HPLC-QTOF-MS: High performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry; ICP-MS: Inductively coupled plasma mass spectrometry; PCA: Principal components analysis; CG: Control group; FAFLG: Low-dosage group of Fructus Aurantii flavonoids; FAFMG: Middle-dosage group of Fructus Aurantii flavonoids; FAFHG: High-dosage group of Fructus Aurantii flavonoids; DPG: Domperidone group.
  2,996 141 5
Acacia catechu ethanolic seed extract triggers apoptosis of SCC-25 cells
Thangavelu Lakshmi, Devaraj Ezhilarasan, Upendra Nagaich, Rajagopal Vijayaragavan
July-September 2017, 13(51):405-411
DOI:10.4103/pm.pm_458_16  PMID:29142391
Background: Acacia catechu Willd (Fabaceae), commonly known as catechu, cachou, and black cutch, has been studied for its hepatoprotective, antipyretic, antidiarrheal, hypoglycemic, anti-inflammatory, immunomodulatory, antinociceptive, antimicrobial, free radical scavenging, and antioxidant activities. Objective: We evaluated the cytotoxic activity of ethanol extract of A. catechu seed (ACS) against SCC-25 human oral squamous carcinoma cell line. Methods: Cytotoxic effect of ACS extract was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, using concentrations of 0.1–1000 μ g/mL for 24 h. A. catechu ethanol seed extract was treated SCC-25 cells with 25 and 50 μ g/mL. At the end of treatment period, apoptotic marker gene expressions such as caspase 8, 9, Bcl-2, Bax, and cytochrome c were evaluated by semiquantitative reverse transcription-polymerase chain reaction. Morphological changes of ACS treated SCC-25 cells was evaluated by acridine orange/ethidium bromide (AO/EB) dual staining. Nuclear morphology and DNA fragmentation was evaluated by propidium iodide (PI) staining. Results: A. catechu ethanol seed extract treatment caused cytotoxicity in SCC-25 cells with an IC50value of 100 μ g/mL. Apoptotic markers caspases 8 and 9, cytochrome c, Bax gene expressions were significantly increased upon ACS extract treatment indicate the apoptosis induction in SCC-25 cells. This treatment also caused significant downregulation of Bcl-2 gene expression. Staining with AO/EB and PI shows membrane blebbing, and nuclear membrane distortion further confirms the apoptosis induction by ACS treatment in SCC-25 cells. Conclusion: The ethanol seed extracts of A. catechu was found to be cytotoxic at lower concentrations and induced apoptosis in human oral squamous carcinoma SCC-25 cells. Abbreviations used: ACS: Acacia catechu seed extract; MTT: 3 (4,5 dimethylthiazol 2 yl) 2,5 diphenyltetrazolium bromide; DMSO: Dimethyl sulfoxide; AO/EO: Acridine orange/ethidium bromide; LC MS: Liquid chromatography mass spectrometry.
  2,816 187 6
Hypoglycemic, hypolipidemic, and wound healing potential of quercetin in streptozotocin-induced diabetic rats
Mahrukh Ahmad, Mudasir Sultana, Rajinder Raina, Nrip Kishore Pankaj, Pawan Kumar Verma, Shahid Prawez
July-September 2017, 13(51):633-639
DOI:10.4103/pm.pm_108_17  PMID:29142425
Background: Among the dietary polyphenolic, quercetin is the most common compound available in vegetables and fruits. The phytochemicals are used to treat diabetic wounds and diabetes, and specifically dietary polyphenols are being extensively studied for their anti-inflammatory and antioxidant abilities. Objective: The objective of the study was to assess the hypoglycemic, hypolipidemic, and wound healing potential of quercetin in streptozotocin (STZ)-induced diabetic Wistar rats. Materials and Methods: Induction of diabetes was done by intraperitoneally administration of STZ at the dose of 55 mg/kg in Wistar rats. An excision wound was created in diabetic rats that were treated with quercetin (100 mg/kg) orally and quercetin ointment topically to evaluate the antidiabetic and wound healing potential of quercetin. Results: Repeated oral administration of quercetin along with topical application of quercetin ointment in diabetic rats normalized the altered blood glucose, hydroxyproline, and glucosamine levels. Topical application of quercetin ointment alone on the excised wound was sufficient enough to heal the wound area in diabetic rats. Conclusions: The result of the present study indicates that quercetin produces hypoglycemic effect in STZ-induced diabetic rats and normalized plasma lipids and protein profiles. Besides, this quercetin also has an excellent wound healing property when applied topically on the wound area in diabetic rats. Abbreviation used: STZ: Streptozotocin; CMC: Carboxy methyl cellulose; HDL: High density lipoproteins; LDL: low density lipoproteins.
  2,754 185 5
Preparation and bioactivity of exopolysaccharide from an endophytic fungus Chaetomium sp. of the medicinal plant gynostemma pentaphylla
Huiru Zhang, Xueqin Wang, Ruifang Li, Xincheng Sun, Siwen Sun, Qiang Li, Chunping Xu
July-September 2017, 13(51):477-482
DOI:10.4103/0973-1296.211033  PMID:28839375
Background: Many exopolysaccharides from the endophytes in medicinal plants possess various potential bioactivities. Materials and Methods: The endophytic fungus JY25 was isolated from the leave of the Chinese medicinal plant Gynostemma pentaphylla and identified as Chaetomium sp. by its phylogenetic and physiological analysis. One exopolysaccharide (EPS) fraction was isolated from the fermentation broth by ethanol precipitation and purified by gel filtration chromatography on Sepharose CL-6B. The molecular characteristics were examined by GC-MS, FT-IR, and multiangle laser light scattering (MALLS). Results: The monosaccharide composition analysis indicated that the purified EPS was mainly composed of glucose, mannose, arabinose, and galactose with the molecular ratio of 78.29: 8.99: 8.64: 4.08. FT-IR spectral analysis of the purified EPS revealed prominent characteristic groups, such as carbonyl bond, pyranose ring, and so on. The weight-average molar mass and the polydispersity ratio of the EPS were revealed to be 1.961×104 g/mol and 1.838, respectively. Furthermore, thermo gravimetric analysis (TGA) indicated that the degradation temperature of the purified EPS was 305°C. The purified EPS from the endophytic fungus Chaetomium sp. displayed antioxidant and antiproliferative activities. Conclusion: The results demonstrated that the EPS could be used as a healthful food and material source in pharmaceutical industries. Abbreviations used: ANOVA: Analysis of variance; DPPH: 2,2-diphenyl-1-picrylhydrazyl; EPS: Exopolysaccharide; FT-IR: Fourier transform infrared spectroscopy; GC-MS: Gas chromatography–mass spectrometry; Mw: Mass weight; MALLS: Multiangle laser light scattering; SEC: Size Exclusion Chomatography; SPSS: Statistical Package of the Social Science; TGA: Thermo gravimetric analysis; TFA: Trifluoroacetic acid.
  2,815 122 6
Antihyperglycemic activity of Caralluma fimbriata: An In vitro approach
Shenai Ashwini, Roy Anitha
July-September 2017, 13(51):499-504
DOI:10.4103/pm.pm_59_17  PMID:29142405
Background: An increase in prevalence of diabetes mellitus necessitates the need to develop new drugs for its effective management. Plants and their bioactive compounds are found to be an alternative therapeutic approach. Caralluma fimbriata, used in this study, is well known for its various biological effects. Objective: The present study was designed to investigate the antihyperglycemic effect of the ethanolic leaf extract of C. fimbriata. Materials and Methods: Different concentrations (1–1000 μ g/mL) of the ethanolic leaf extract of C. fimbriata were subjected to alpha-amylase and alpha-glucosidase inhibitory assay with acarbose as control. Cytotoxicity was assessed by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Glucose uptake assay was performed on L6 myotubes using the extract in 1 μ g–100 μ g/mL, using metformin and insulin as control. Results: The C. fimbriata extract showed potent inhibitory activity on enzymes of glucose metabolism in a dose-dependent manner. The maximum alpha-amylase inhibitory effect was 77.37% ± 3.23% at 1000 μ g/mL with an IC50value of 41.75 μ g/mL and alpha-glucosidase inhibitory effect was 83.05% ± 1.69% at 1000 μ g/mL with an IC50value of 66.71 μ g/mL. The maximum glucose uptake was found to be 66.32% ± 0.29% for the Caralluma extract at 100 μ g/mL and that of metformin (10 μ g/mL) was 74.44% ± 1.72% and insulin (10 μ M) 85.55% ± 1.14%. The extract was found to be safe as the IC50of extract and metformin was found to be ≥1000 μ g/mL and ≥1000 μ M, respectively, in the cell line tested. Conclusion: The study concludes that C. fimbriata has promising antihyperglycemic activity. Abbreviations used: GLUT: Glucose transporter; MTT: 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide.
  2,707 203 1
Optimization of polysaccharide ultrasonic extraction conditions using purple sweet potato tubers based on free radical scavenging and glycosylation inhibitory bioactivities
Haihua Guo, Fansheng Kong, Chunyan Yan
July-September 2017, 13(51):504-511
DOI:10.4103/0973-1296.211044  PMID:28839380
Background: The purple sweet potato, Ipomoea batatas, belongs to the family Convolvulaceae. It is one of the most widely consumed tubers in Asia and is found in many dishes. Many people with diabetes eat purple sweet potato tubers to help reduce blood glucose in China. Objective: To predict the ultrasonic conditions for getting the optimal in vitro antioxidant and antiglycated activity of ultrasonic extracted polysaccharides from purple sweet potato (I. batatas) tubers, the artificial neural network (ANN) regression models was used in this study. Materials and Methods: The antioxidant activity of polysaccharides was quantified by evaluating the hydroxyl radical scavenging effect after ultrasonic extraction, and the data were used in conjunction with optimized extraction conditions to train the predictive ANN models. Results: The following conditions were predicted to yield optimal hydroxyl scavenging activity: 200 W, 22°C, and 40 min. In contrast, conditions of 230 W, 22°C, and 50 min yielded the greatest inhibitory effect on albumin nonenzymatic glycosylation. The accuracy and predictive ability of the models ranged from good to excellent, as indicated by R2 values ranging from 0.953 to 0.998. Conclusion: The results of the present study showed that ANN predictive models are useful in ultrasonic processing, which can rapidly and accurately predict the optimum extraction conditions for polysaccharides based on their antioxidant and antiglycated activities. In addition, the results of the present study suggest that the consumption of sweet potatoes may help reduce free radicals in the body and prevent or treat diabetes. Abbreviations used: IBP: Polysaccharide of Ipomoea batatas; RSM: response surface methodology; ANN: Artificial neural network; BSA: Bovine serum albumin.
  2,743 133 3
A novel quantitative prediction approach for astringency level of herbs based on an electronic tongue
Xue Han, Hong Jiang, Dingkun Zhang, Yingying Zhang, Xi Xiong, Jiaojiao Jiao, Runchun Xu, Ming Yang, Li Han, Junzhi Lin
July-September 2017, 13(51):492-497
DOI:10.4103/pm.pm_455_16  PMID:28839378
Background: The current astringency evaluation for herbs has become dissatisfied with the requirement of pharmaceutical process. It needed a new method to accurately assess astringency. Methods: First, quinine, sucrose, citric acid, sodium chloride, monosodium glutamate, and tannic acid (TA) were analyzed by electronic tongue (e-tongue) to determine the approximate region of astringency in partial least square (PLS) map. Second, different concentrations of TA were detected to define the standard curve of astringency. Meanwhile, coordinate-concentration relationship could be obtained by fitting the PLS abscissa of standard curve and corresponding concentration. Third, Chebulae Fructus (CF), Yuganzi throat tablets (YGZTT), and Sanlejiang oral liquid (SLJOL) were tested to define the region in PLS map. Finally, the astringent intensities of samples were calculated combining with the standard coordinate-concentration relationship and expressed by concentrations of TA. Then, Euclidean distance (Ed) analysis and human sensory test were processed to verify the results. Results: The fitting equation between concentration and abscissa of TA was Y = 0.00498 × e(−X/0.51035) + 0.10905 (r = 0.999). The astringency of 1, 0.1 mg/mL CF was predicted at 0.28, 0.12 mg/mL TA; 2, 0.2 mg/mL YGZTTs was predicted at 0.18, 0.11 mg/mL TA; 0.002, 0.0002 mg/mL SLJOL was predicted at 0.15, 0.10 mg/mL TA. The validation results showed that the predicted astringency of e-tongue was basically consistent to human sensory and was more accuracy than Edanalysis. Conclusion: The study indicated the established method was objective and feasible. It provided a new quantitative method for astringency of herbs. Abbreviations used: CF: Chebulae Fructus; E-tongue: Electronic tongue; Ed: Euclidean distance; PLS: Partial least square; PCA: Principal component analysis; SLJOL: Sanlejiang oral liquid; TA: Tannic acid; VAS: Visual analog scale; YGZTT: Yuganzi throat tablets.
  2,722 145 4
Chalepin: A compound from Ruta angustifolia L. pers exhibits cell cycle arrest at S phase, suppresses nuclear factor-kappa B (NF-κB) pathway, signal transducer and activation of transcription 3 (STAT3) phosphorylation and extrinsic apoptotic pathway in non-small cell lung cancer carcinoma (A549)
Jaime Stella Moses Richardson, Norhaniza Aminudin, Sri Nurestri Abd Malek
July-September 2017, 13(51):489-498
DOI:10.4103/pm.pm_13_17  PMID:29142404
Background: Plants have been a major source of inspiration in developing novel drug compounds in the treatment of various diseases that afflict human beings worldwide. Ruta angustifolia L. Pers known locally as Garuda has been conventionally used for various medicinal purposes such as in the treatment of cancer. Objective: A dihydrofuranocoumarin named chalepin, which was isolated from the chloroform extract of the plant, was tested on its ability to inhibit molecular pathways of human lung carcinoma (A549) cells. Materials and Methods: Cell cycle analysis and caspase 8 activation were conducted using a flow cytometer, and protein expressions in molecular pathways were determined using Western blot technique. Results: Cell cycle analysis showed that cell cycle was arrested at the S phase. Further studies using Western blotting technique showed that cell cycle-related proteins such as cyclins, cyclin-dependent kinases (CDKs), and inhibitors of CDKs correspond to a cell cycle arrest at the S phase. Chalepin also showed inhibition in the expression of inhibitors of apoptosis proteins. Nuclear factor-kappa B (NF-κB) pathway, signal transducer and activation of transcription 3 (STAT-3), cyclooxygenase-2, and c-myc were also downregulated upon treatment with chalepin. Chalepin was found to induce extrinsic apoptotic pathway. Death receptors 4 and 5 showed a dramatic upregulation at 24 h. Analysis of activation of caspase 8 with the flow cytometer showed an increase in activity in a dose- and time-dependent manner. Activation of caspase 8 induced cleavage of BH3-interacting domain death agonist, which initiated a mitochondrial-dependent or -independent apoptosis. Conclusion: Chalepin causes S phase cell cycle arrest, NF-κB pathway inhibition, and STAT-3 inhibition, induces extrinsic apoptotic pathway, and could be an excellent chemotherapeutic agent. Abbreviations used: °C: Degree Celsius, ANOVA: Analysis of variance, ATCC: American Type Culture Collection, BCL-2: B-Cell CLL/Lymphoma 2, Bcl-xL: B-cell lymphoma extra-large, BH3: Bcl-2 homology 3, BID: BH3-interacting domain death agonist, BIR: Baculovirus inhibitor of apoptosis protein repeat, Caspases: Cysteinyl aspartate-specific proteases, CDK: Cyclin-dependent kinase, CO2: Carbon dioxide, CST: Cell signaling technologies, DISC: Death-inducing signaling complex, DMSO: Dimethyl sulfoxide, DNA: Deoxyribonucleic acid, DR4: Death receptor 4, DR5: Death receptor 5, E1a: Adenovirus early region 1A, ECL: Enhanced chemiluminescence, EDTA: Ethylenediaminetetraacetic acid, ELISA: Enzyme-linked immunosorbent assay, etc.: Etcetera, FADD: Fas-associated protein with death domain, FBS: Fetal bovine serum, FITC: Fluorescein isothiocyanate, G1: Gap 1, G2: Gap 2, HPLC: High-performance liquid chromatography, HRP: Horseradish peroxidase, IAPs: Inhibitor of apoptosis proteins, IC50: Inhibitory concentration at half maximal inhibitory, IKK-α : Inhibitor of nuclear factor kappa-B kinase subunit alpha, IKK-β : Inhibitor of nuclear factor kappa-B kinase subunit beta, IKK-γ : Inhibitor of nuclear factor kappa-B kinase subunit gamma, IKK: IκB kinase, IκBα : Nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha, m: Meter, M: Mitotic, mm: Millimeter, mRNA: Messenger ribonucleic acid, NaCl: Sodium chloride, NaVO4: Sodium orthovanadate, NEMO: NF-Kappa-B essential modulator, NF-κB: Nuclear factor kappa-light chain-enhancer of activated B cells, NSCLC: Nonsmall cell lung carcinoma, PBS: Phosphate buffered saline, PGE2: Prostaglandin E2, PI: Propidium iodide, PMSF: Phenylmethylsulfonyl fluoride, pRB: Phosphorylated retinoblastoma, R. angustifolia: Ruta angustifolia L. Pers, Rb: Retinoblastoma, rpm: Rotation per minute, RPMI: Roswell Park Memorial Institute, S phase: Synthesis phase, SD: Standard deviation, SDS-PAGE: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Smac: Second mitochondria-derived activator of caspase, SPSS: Statistical Package for the Social Sciences, STAT3: Signal transducer and activation of transcription 3, tBID: Truncated BID, TNF: Tumor necrosis factor, TRADD: Tumor necrosis factor receptor type-1 associated death domain, TRAIL: TNF-related apoptosis-inducing ligand, USA: United States of America, v/v: Volume over volume.
  2,635 182 3
Simultaneous quantitation of six aconitum alkaloids and three flavonoids in the herb couple of radix aconiti lateralis-radix glycyrrhizae (Fuzi–Gancao) by UHPLC-ESI-MS/MS
Xiao Fu, Rongrong Lu, Songsong Zhao
July-September 2017, 13(51):425-429
DOI:10.4103/pm.pm_141_16  PMID:28839367
Background: Fuzi–Gancao herb couple is one of the commonly used herb couples involved in the traditional Chinese medicine formulations, with radix aconiti lateralis (Fuzi in Chinese) and radix glycyrrhizae (Gancao in Chinese) as a ratio of 1:1. Alkaloids and flavonoids were considered as the main active ingredients of Fuzi–Gancao herb couple. However, no analytical methods have been reported to quantitatively analyze these activity ingredients in Fuzi–Gancao herb couple simultaneously. Objective: To develop a simple, rapid, and sensitive UHPLC-MS/MS method for simultaneous quantitation of six alkaloid and three flavonoid compounds, namely, aconitine (AC), mesaconitine (MA), hypaconitine (HA), benzoylaconitine (BAC), benzoylmesaconitine (BMA), benzoylhypaconitine (BHA), liquiritin, isoliquiritin, and licochalcone A (LCA) in Fuzi–Gancao herb couple. Materials and Methods: The chromatographic separation was achieved using a reversed phase C18 column and a mobile phase consisted of water (0.1% formic acid in water, v/v) and acetonitrile. The detection was achieved in multiple reaction monitoring modes. The optimal mass transition ion pairs (m/z) for quantitation were 646.3/586.5 for AC, 632.4/572.5 for MA, 616.3/556.4 for HA, 604.5/572.5 for BAC, 590.4/540.4 for BMA, 574.1/542.5 for BHA, 419.4/257.1 for liquiritin, 419.4/257.1 for isoliquiritin, and 339.2/121.1 for LCA. Results: Good linearity was observed in validated concentration range for each analyte (r > 0.9992), and the intra- and inter-day precisions were <3.12% and 3.65%, respectively. The recovery ranged from 85.36% to 110.14%. Conclusions: The results demonstrated that the method developed was reliable, rapid, and specific. Moreover, this method was successfully applied to control the quality of Fuzi–Gancao herb couple. Abbreviations used: UHPLC-ESI-MS/MS: Ultra High Performance Liquid Chromatography-electrospray ionization-mass spectrometry; RSD: Relative standard deviations; LOD: Limit of detection; LOQ: Limit of quantification; MRM: Multiple reaction monitor; TCMs: Traditional Chinese medicines.
  2,678 121 3
Antitumor effects and mechanism of n-butanol fraction from aril of Torreya grandis in H22 mice
Fang-fang Duan, Shan-shan Jia, Ke Yuan
July-September 2017, 13(51):351-357
DOI:10.4103/pm.pm_286_16  PMID:28839356
Background: To determine the antitumor effects and its mechanism of n-butanol fraction from aril of Torreya grandis (BFAT) on H22mice models of liver cancer. Materials and Methods: Sixty ICR male mice were used to establish H22mice models of liver cancer and then randomly divided into six groups, the normal control group, the model control group, the positive group (cyclophosphamide [CTX]), the BFAT-treated group (high, 4 g/kg, medium, 2 g/kg, and low, 1 g/kg). The animals were sacrificed 15 days after oral administration, and tumors were taken out for the tumor weights and antitumor rates, while thymus and spleen were taken for thymus index and spleen index. Blood in eyeball was collected for the determination of aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin (Alb), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase enzyme (GSH-Px), tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), transforming growth factor-β1 (TGF-β1), and IL-10 in serum. Sections of tumor tissue were prepared, and morphological changes in tumor tissue cells were observed using hematoxylin and eosin staining technique. Results: Compared with the model control group, the tumor inhibition rate of the high-dose administered group is 60.15%, which is quite closed to the effect of CTX. Moreover, the tumor weight is decreased, the indexes of spleen, thymus were increased significantly. Furthermore, the administration of BFAT significantly enhanced the activities of TNF-α, IL-2, SOD, and GSH-Px and reduced the levels of AST, ALT, MDA, Alb, TGF-β1, and IL-10 (P < 0.01). Conclusions: The results demonstrated that n-butanol fraction from aril of T. grandis showed out antitumor activity without obviously liver damage through potentiating immunologic function and antioxidant activity of tumor-bearing mice and which may become one potential as anticancer drug alternatives or supplements. Abbreviations used: DDP: Cisplatin; CTX: Cyclophosphamide; AST: Aspartate aminotransferase; ALT: Alanine aminotransferase; Alb: Albumin; MDA: Malondialdehyde; SOD: Superoxide dismutase; GSH-Px: Glutathione peroxide enzyme; TNF-α: Tumor necrosis factor-α; IL-2: Interleukin-2; TGF-α1: Transforming growth factor-α1; IL-10: Interleukin-10; HE: Hematoxylin and eosin; PBS: Phosphate- buffered saline; PFAT: Petroleum ether fraction from aril of Torreya grandis; EFAT: Ethyl acetate fraction from aril of Torreya grandis; BFAT: N-butanol fraction from aril of Torreya grandis.
  2,669 104 1
Characterization of nano bamboo charcoal drug delivery system for Eucommia ulmoides extract and its anticancer effect In vitro
Zhaoyan Zeng, Xiangzhou Li, Sheng Zhang, Dan Huang
July-September 2017, 13(51):498-503
DOI:10.4103/pm.pm_256_16  PMID:28839379
Background: Nano bamboo charcoal is being widely used as sustained release carrier for chemicals for its high specific surface area, sound biocompatibility, and nontoxicity; however, there have been no reports on nano bamboo charcoal as sustained release carrier for traditional Chinese medicine (TCM). Objective: To study the effect of nano bamboo charcoal in absorbing and sustained releasing Eucommia ulmoides extract (EUE) and to verify the in vitro anticancer effect of the sustained release liquid, so as to provide a theoretical basis for the development and utilization of nano bamboo charcoal as TCM sustained-release preparation. Materials and Methods: The adsorption capacity for the nano bamboo charcoal on EUE was measured by Langmuir model, and the release experiment was carried out under intestinal fluid condition. Characteristic changes for the nano bamboo charcoal nano-drug delivery system with and without adsorption of E. ulmoides were evaluated by scanning electron microscopy, Fourier transform infrared spectroscopy, thermogravimetric analysis, and specific surface area. In addition, the anticancer effect from this novel bamboo charcoal E. ulmoides delivery system was evaluated against a human colon cancer cell line (HCT116). Results: It was found that nano bamboo charcoal exhibits good adsorption capacity (up to 462.96 mg/g at 37°C). The cumulative release rate for EUE from this nano bamboo charcoal delivery system was 70.67%, and specific surface area for the nano bamboo charcoal decreased from 820.32 m2/g to 443.80 m2/g after EUE was loaded. An in vitro anticancer study showed that the inhibition rate for E. ulmoides against HCT116 cancer cells was 23.07%, for this novel bamboo charcoal nano-drug delivery system. Conclusion: This study provides a novel strategy for the delivery of traditional Chinese medicine using bamboo charcoal nano-drug delivery system. Abbreviation used: EUE: Eucommia ulmoides extract.
  2,640 131 7
Apoptotic effect of Nigella sativa on human lymphoma U937 cells
Belkis Atasever Arslan, Fatma Busra Isik, Hazal Gur, Fatih Ozen, Tunc Catal
July-September 2017, 13(51):628-632
DOI:10.4103/pm.pm_93_17  PMID:29142424
Objective: Nigella sativa is from botanical Ranunculaceae family and commonly known as black seed. Apoptotic effect of N. sativa and its apoptotic signaling pathways on U937 lymphoma cells are unknown. Materials and Methods: In this study, we investigated selective cytotoxic and apoptotic effects of N. sativa extract and its apoptotic mechanisms on U937 cells. In addition, we also studied selective cytotoxic activity of thymoquinone that is the most active essential oil of N. sativa. Results: Our results showed that N. sativa extract has selective cytotoxicity and apoptotic effects on U937 cells but not ECV304 control cells. However, thymoquinone had no significant cytotoxicity against on both cells. N. sativa extract increased significantly caspase-3, BAD, and p53 gene expressions in U937 cells. Conclusions: N. sativa may have anticancer drug potential and trigger p53-induced apoptosis in U937 lymphoma cells. Abbreviations used: CI: Cytotoxicity index, DMEM: Dulbecco's Modified Eagle Medium, HL: Hodgkin's lymphoma, MTT: 3-(4,5-dimethy lthiazol-2yl)-2,5-diphenyl tetrazolium bromide, RPMI: Roswell Park Memorial Institute medium.
  2,618 134 1
Molecular docking analysis of phytic acid and 4-hydroxyisoleucine as cyclooxygenase-2, microsomal prostaglandin E synthase-2, tyrosinase, human neutrophil elastase, matrix metalloproteinase-2 and -9, xanthine oxidase, squalene synthase, nitric oxide synthase, human aldose reductase, and lipoxygenase inhibitors
Radhakrishnan Narayanaswamy, Lam Kok Wai, Norhaizan Mohd Esa
July-September 2017, 13(51):512-518
DOI:10.4103/pm.pm_195_16  PMID:29142407
Background: The phytoconstituents phytic acid and 4-hydroxyisoleucine have been reported to posses various biological properties. Objective: This prompted us to carry out the docking study on these two ligands (phytic acid & 4-hydroxyisoleucine) against eleven targeted enzymes. Materials and Methods: Phytic acid & 4-hydroxyisoleucine were evaluated on the docking behaviour of cyclooxygenase-2 (COX-2), microsomal prostaglandin E synthase-2 (mPGES-2), tyrosinase, human neutrophil elastase (HNE), matrix metalloproteinase (MMP 2 and 9), xanthine oxidase (XO), squalene synthase (SQS), nitric oxide synthase (NOS), human aldose reductase (HAR) and lipoxygenase (LOX) using Discovery Studio Version 3.1 (except for LOX, where Autodock 4.2 tool was used). Results: Docking and binding free energy analysis revealed that phytic acid exhibited the maximum binding energy for four target enzymes such as COX-2, mPGES-2, tyrosinase and HNE. Interestingly, we found that 4-hydroxyisoleucine has the potential to dock and bind with all of the eleven targeted enzymes. Conclusion: This present study has paved a new insight in understanding 4-hydroxyisoleucine as potential inhibitor against COX-2, mPGES-2, tyrosinase, HNE, MMP 2, MMP 9, XO, SQS, NOS, HAR and LOX. Abbreviations used: COX-2: Cyclooxygenase-2, mPGES-2: Microsomal prostaglandin E synthase-2, HNE: Human neutrophil elastase, MMP-2 and -9: Matrix metalloproteinase-2 and -9, XO: Xanthine oxidase, SQS: Squalene synthase, NOS: Nitric oxide synthase, HAR: Human aldose reductase, LOX: Lipoxygenase, ADME: Absorption, distribution, metabolism, and excretion, TOPKAT: Toxicity Prediction by Computer-assisted Technology.
  2,585 165 3
Sugemule-3 protects against isoprenaline-induced cardiotoxicity In vitro
Yu Wang, Guo-Hua Gong, Ya-Nan Xu, Li-Jun Yu, Cheng-Xi Wei
July-September 2017, 13(51):517-522
DOI:10.4103/0973-1296.211018  PMID:28839382
Background: Sugemule-3 (SD) is a traditional Chinese medicine with protective effect of myocardium. However, the underlying mechanisms of the effect had not been elucidated. Materials and Methods: In the present study, the serum of SD was prepared. A model of β-adrenergic agonist isoprenaline (ISO)-induced H9c2 cardiomyocytes injury was established in vitro. The changes in cell viability were examined to determine the available concentration of ISO and serum of SD. ELISA, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay, and flow cytometry were used to detect the effect of serum of SD on oxidative stress and apoptosis. The expression levels of the mitochondria-dependent apoptotic pathway and mitogen-activated protein kinase signalling-related proteins were analyzed. Results: Incubation with different dose of ISO (0.015, 0.01, 0.005, and 0.0025 mol/L) for 24 h caused dose-dependent loss of cell viability and 0.01 mol/L of ISO approximately reduced the cell viability to 50%. Pretreatment with 50 μ mol/L serum of SD effectively decreased the levels of ISO-induced cell toxicity. Serum of SD relived ISO-induced oxidative stress and apoptosis in H9c2 cardiomyocytes. A further mechanism study indicated that serum of SD inhibited the mitochondria-dependent apoptotic pathways and regulated the expression levels of Bcl-2 family. ISO activated ERK and P38, whereas serum of SD inhibited their activation. Conclusion: Serum of SD inhibits the ISO-induced activation of the mitochondria-dependent apoptotic pathway, oxidative stress, and ERK, P38 inactivation. Serum of SD is used for the treatment of ISO-induced cardiomyopathy. Abbreviations used: ELISA: Enzyme-linked Immunosorbent Assay; TUNEL: TdT-mediated dUTP nick end labeling; MTT: 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, DMSO: dimethyl sulfoxide; MDA: Malondialdehyde; SOD: Superoxide Dismutase; GSH-Px: Glutathione peroxidase.
  2,635 112 4
Antioxidant and cholinesterase inhibitory activities of ethyl acetate extract of Terminalia chebula: Cell-free In vitro and In silico studies
Mohamed Asik Rajmohamed, Suganthy Natarajan, Premkumar Palanisamy, Akbarsha Mohammad Abdulkader, Archunan Govindaraju
July-September 2017, 13(51):437-445
DOI:10.4103/pm.pm_57_17  PMID:29142396
Background: Alzheimer's disease (AD) is a progressive neurodegenerative disorder clinically characterized by memory loss and impaired cognitive function. Cholinergic enzyme deficiency and oxidative stress are the two major factors implicated in the pathogenesis of AD. The symptomatic treatment, as of now, is the use of cholinesterase inhibitors toward cholinergic “downturn.” Therefore, there is a search for compounds that will be useful in focused therapies. There has been suggestion that Terminalia chebula fruit would be a potential source. Objective: To assess the anticholinesterase and antioxidant activities of T. chebula fruit which is widely practiced in the Ayurvedic medicines for memory enhancement. Materials and Methods: Ethyl acetate extract of T. chebula fruit (TCEA) was subjected to phytochemical investigation of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitory activities and cell-free antioxidant activity. TCEA was further subjected to gas chromatography-mass spectrum (GC-MS) analysis. The bioactive compounds were analyzed for molecular docking with AChE and BuChE proteins. Results: TCEA exhibited potent AChE and BuChE inhibitory activities comparable to the standard drug donepezil. In vitro cell-free antioxidant assays demonstrated that TCEA possesses excellent free radical scavenging activity, reducing power, and potent metal-chelating activity. Total polyphenolic content of TCEA was 596.75 ± 0.35 μg gallic acid equivalents/mg of extract, which correlates with the antioxidant activity of TCEA. Molecular docking of compounds expounded in GC-MS analysis for AChE and BuChE enzyme activities revealed that methyl N-(N-benzyloxycarbonyl-beta-l-aspartyl)-beta-d-glucosaminide as the most potent compound with good predicted activities. Conclusion: Overall, the results revealed that the bioactive molecule methyl N-(N-benzyloxycarbonyl-beta-l-aspartyl)-beta-d-glucosaminide present in TCEA is a potential depressant for the treatment of AD and related neurodegenerative disorders. Abbreviations used: AD: Alzheimer's disease; TCEA: Ethyl acetate extract of Terminalia chebula; GC-MS: Gas chromatography-mass spectrum; ROS: Reactive oxygen species; RNS: Reactive nitrogen species; AChE: Acetylcholinesterase; BuChE: Butyrylcholinesterase; NFT: Neurofibrillary tangles; Aβ : β -amyloid; NSAIDS: Nonsteroidal anti-inflammatory drugs; FDA: Food and Drug Administration; RT: Room temperature; HCl: Hydrochloric acid; ATCI: Acetylthiocholine iodide; BTCI: Butyrylthiocholine iodide; BHT: Butylated hydroxytoluene; DPPH: 2,2-diphenyl-1-picrylhydrazyl; TCA: Trichloroacetic acid; GAE: Gallic acid equivalent; NICT: National Institute of Information and Communications Technology; 3D: Three-dimensional; PDB: Protein data bank; OPLS: Optimized potentials for liquid simulations; XP: Extra precision; SD: Standard deviation; ANOVA: Analysis of variance; EDTA: Ethylenediaminetetraacetic acid.
  2,556 176 1
Decoction of Dictamnus Dasycarpus Turcz. Root bark ameliorates skin lesions and inhibits inflammatory reactions in mice with contact dermatitis
Beodeul Yang, Hong-Bum Lee, Sura Kim, Young Chul Park, Koanhoi Kim, Hyungwoo Kim
July-September 2017, 13(51):483-487
DOI:10.4103/0973-1296.211034  PMID:28839376
Background: The root bark of Dictamnus dasycarpus Turcz. (Dictamni Radicis Cortex) has been widely used to treat skin diseases in Korea, and its anti-inflammatory efficacies were recently reported. Objective: The paper aims to investigate the inhibitory effects of decoction of Dictamni Radicis Cortex (DDRC) in mice with contact dermatitis (CD). Materials and Methods: We investigated the effects of DDRC on skin lesion characteristics such as crust, scales, incrustation and petechiae, the erythema and melanin indexes, skin thickness, histopathologic changes, and cytokine production in 1-fluoro-2,4-dinitrofluorobenzene (DNFB)-induced CD mice. Results: Topical application of DDRC ameliorated crust, scales, incrustation, and induced by DNFB. In addition, DDRC lowered the erythema index significantly (P < 0.05). DDRC effectively inhibited enlargement of skin thickness (P < 0.05). Histopathologic observation showed that DDRC inhibited epidermal hyperplasia, hyperkeratosis, and spongiotic changes. Finally, DDRC decreased production levels of IFN-γ, TNF-α and IL-6 induced by repeated application of DNFB (P < 0.05). Conclusion: These data suggest that DDRC can be used in the treatment of inflammatory skin diseases including CD. Moreover, these results are closely related to the decreasing production of TNF-α IFN-γ and IL-6 in inflamed tissues. Abbreviations used: DDRC: decoction of Dictamni Radicis Cortex, CD: contact dermatitis, DNFB: 1-fluoro-2,4-dinitrofluorobenzene, AOO: acetone and olive oil, DEX: dexamethasone, CBA: cytometric bead array
  2,590 139 6
Tyrosinase inhibitory activities of Carissa opaca Stapf ex haines roots extracts and their phytochemical analysis
Wajeeha Malik, Dildar Ahmed, Sania Izhar
July-September 2017, 13(51):544-548
DOI:10.4103/pm.pm_561_16  PMID:29142412
Objective: Carissa opaca is a medicinal plant with rich folkloric applications. The present research was conducted to explore the tyrosinase inhibitory potential of aqueous decoction (AD) and methanolic extract (ME) of roots of C. opaca and its fractions in various solvents and their phytochemical analysis. Materials and Methods: AD of the dried powdered roots of C. opaca was prepared by boiling in water. ME was prepared by cold maceration. Its fractions were obtained in solvents of increasing polarity, i.e., hexane, chloroform, ethyl acetate, n-butanol, and water. The biomass left after extraction with methanol was boiled in water to get its decoction Biomass aqueous decoction (BAD). Tyrosinase inhibitory activities of the samples were studied according to a reported method. Chemical compounds in the samples were identified by gas chromatography-mass spectrometry (GC-MS). Results: The AD, BAD, and ME and its fractions displayed remarkable tyrosinase inhibitory activity. The IC50of AD was 23.33 μ g/mL as compared to 15.80 μ g/mL of the standard arbutin and that of BAD was 21.24 μ g/mL. The IC50of ME was 34.76 μ g/mL while that of hexane, chloroform, ethyl acetate, n-butanolic, and aqueous fractions was 21.0, 44.73, 43.40, 27.66, and 25.06 μ g/mL, respectively. The hexane fraction was thus most potent followed by aqueous fraction. By phytochemical analysis, campesterol, stigmasterol, gamma-sitosterol, alpha-amyrin, 9,19-cyclolanostan-3-ol, 24-methylene-,(3 β )-, lupeol, lup-20(29)-en-3-one, lup-20(29)-en-3-ol, acetate,(3 β )-, 2(1H) naphthalenone, 3,5,6,7,8,8a-hexahydro-4,8a-dimethyl-6-(1-methylethenyl)-, and 2,3,3-trimethyl-2-(3-methylbuta-1,3-dienyl)-6-methylenecyclohexanone were identified in the extracts by GC-MS. Other compounds included fatty acids and their esters. Some of these compounds are being first time reported here from this plant. Conclusions: The roots extracts exhibited considerable tyrosinase inhibitory activities, alluding to a possible application of the plant in cosmetic as whitening agent subject to further pharmacological studies. Abbreviations used: AD: Aqueous decoction; ME: Methanolic extract; BAD: Biomass aqueous decoction; GC-MS: Gas chromatography-mass spectrometry.
  2,632 85 1
Anti-epileptic effects of valepotriate isolated from Valeriana jatamansi jones and its possible mechanisms
An Wu, Xia Ye, Qiang Huang, Wei-Min Dai, Jian-Min Zhang
July-September 2017, 13(51):512-516
DOI:10.4103/0973-1296.211027  PMID:28839381
Objective: This work aimed to investigate the anti-epileptic effects of valepotriate isolated from Valeriana jatamansi Jones and studied its possible mechanisms. Methods: The anti-epileptic effects of valepotriate were studied using maximal electroshock-induced seizure (MES), pentylenetetrazole (PTZ)-induced epilepsy, and pentobarbital sodium-induced sleeping model in mice. The possible anti-epileptic mechanisms of valepotriate were investigated by analyzing the expressions of GABAA, GABAB, glutamic acid decarboxylase (GAD65), Bcl-2, and caspase-3 in the brain using Western blot assay. Results: The results indicated that valepotriate showed significant anti-epileptic activity against MES- and PTZ-induced epilepsy at doses of 5, 10, and 20 mg/kg, and ED50values for MES- and PTZ-induced epilepsy were 7.84 and 7.19 mg/kg, respectively. Furthermore, valepotriate (10 and 20 mg/kg) can significantly prolong sleeping time and shorten the latency time on the pentobarbital sodium-induced sleeping time test. Furthermore, valepotriate (5, 10, and 20 mg/kg) could significantly up-regulate the expression of GABAA, GAD65, and Bcl-2 and down-regulate the expression of caspase-3, but had no significant effect on the expression of GABAB. Conclusion: The results indicated that valepotriate had anti-epileptic activity and the mechanisms might be associated with regulation of GABA and inhibition of neuronal apoptosis.
  2,583 119 3
Ethanolic fraction of Terminalia tomentosa attenuates biochemical and physiological derangements in diet induced obese rat model by regulating key lipid metabolizing enzymes and adipokines
Balaji Meriga, Parim Brahma Naidu, Ganjayi Muniswamy, GEN Hanuma Kumar, Ramavat Ravindar Naik, Suresh Pothani
July-September 2017, 13(51):385-392
DOI:10.4103/0973-1296.208871  PMID:28839361
The prevalence of overweight-obesity and associated comorbidities have reached alarming levels necessitating the need to explore effective therapeutics. In the present work, we demonstrated the promising antiobesity activity of ethanolic fraction of Terminalia tomentosa bark (EFTT) in diet induced obese rat model. High Fat Diet (HFD)-fed obese rats were orally administered with EFTT (50, 100 and 200 mg/kg body weight). Changes in body weight, body composition, bone mineral concentration, bone mineral density, plasma glucose, insulin, leptin, adiponectin, circulatory and tissue lipid profiles, and the activities of liver antioxidant enzymes, key lipid metabolic enzymes and mRNA expressions of fatty acid synthase (FAS), peroxisome proliferator-activated receptor gamma (PPAR-γ), leptin and tumor necrosis factor alpha (TNF-α) were assessed in experimental rats in the presence and absence of EFTT. At a dose of 200 mg/kg b.wt, EFTT has substantially attenuated body weight and related patho-physiological alterations in HFD-induced obese rats. These findings were correlated with histological observations of adipose tissue. The therapeutic activity of EFTT could be possibly through restoration of antioxidants status, regulation of key lipid metabolizing enzymes, expression of FAS, leptin, PPAR-γ and by synchronized control of energy metabolism in liver and adipose tissue.
  2,523 122 1
Intervention effects of atorvastatin combined with Panax notoginseng saponins on rats with atherosclerosis complicated with hepatic injury
Qing-Fang Jiang, Min-Yi Huang, Kang-Yuan Wu, Jie-Ling Weng, Rong-Gui Deng, Xin-Jie Xu, Jian-Pei Xu, Tao Jiang
July-September 2017, 13(51):430-438
DOI:10.4103/pm.pm_424_16  PMID:28839368
Background: Statins cannot be used for some active liver diseases, which limits its application to some extent. The combined use of statins with other drugs may be one of the ways to solve this dilemma. Objective: This research aims to evaluate the effects of atorvastatin combined with Panax notoginseng saponins (PNS) on rats with atherosclerosis (AS) complicated with hepatic injury. Materials and Methods: Seventy-two male Wistar rats were randomly categorized into control group (without any intervention, Group A) and AS model groups, which were divided into hepatic injury (Groups B–E) and nonhepatic injury (Groups F–I) groups. Hepatic and nonhepatic injury groups were intragastrically treated with 5.5 mg/kg·d atorvastatin (Group B, F), 200 mg/kg·d PNS (Group C, G), 5.5 mg/kg·d atorvastatin + 200 mg/kg·d PNS (Group D, H), and normal saline (Group E, I). After 8 weeks, total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol, low density lipoprotein-cholesterol (LDL-C), and serum calcium were analyzed to evaluate the hypolipidemic effect. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase, total bilirubin, and r-glutamyltransferase levels were measured to assess liver function. The thoracic aortas were used for hematoxylin–eosin staining. Results: In both hepatic injury and nonhepatic injury groups, TC, TG and LDL-C levels significantly decreased in Groups B, D, F, and H. ALT and AST levels significantly increased in Group B, but significantly decreased in Groups C and D. The aortic intima thickness was significantly lower in Groups B, D, F, and H than that in the normal saline group. Conclusion: The combination of atorvastatin and PNS treatment showed a significant hypolipidemic effect and hepatic enzyme stability function. Abbreviations used: PNS: Panax notoginseng saponins; AS: Atherosclerosis; TC: Total cholesterol; TG: Triglyceride; HDL-C: High density lipoprotein-cholesterol; LDL-C: Low density lipoprotein-cholesterol; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; ALP: Alkaline phosphatase; T-BIL: Total bilirubin; r-GT: R-glutamyltransferase; HE: Hematoxylin–eosin.
  2,532 111 1
Molecular simulation-based combinatorial modeling and antioxidant activities of zingiberaceae family rhizomes
Talambedu Usha, Sushen Pradhan, Arvind Kumar Goyal, Shanmugarajan Dhivya, HP Prashanth Kumar, Manoj Kumar Singh, Neelu Joshi, Bharat Chandra Basistha, KR Siddalinga Murthy, Saravanakumar Selvaraj, Sushil Kumar Middha
July-September 2017, 13(51):715-722
DOI:10.4103/pm.pm_82_17  PMID:29142438
Objective: The main aim of this scientific report was to investigate a series of phytochemicals in silico and the pharmacology of four plants found at higher altitude in the ginger family, Zingiberaceae (incl. Costaceae) from North-East India, particularly Sikkim. First, the goal was to determine the biological activities of the four herbs (used under Zingiberaceae family) using antioxidant assays to identify the best species. Second, previously reported compounds in litero were subsequently screened for their anticancerous activities using in silico methods. Materials and Methods: Using the methanolic extracts of herbs, quantitative detection of phytochemicals such as total phenols and total flavonoids was detected, and the free radical scavenging activity was also studied using 2,2-diphenyl-1-picryl-hydrazyl (DPPH) assay. Docking process was studied, using Discovery Studio version 3.5, to identify suitable molecules at the protein-binding sites through annealing and genetic simulation algorithms. Grids centered on active sites were obtained with spacing of 54 × 55 × 56, and 0.503 grid spacing was calculated. The methods adopted and used in this study were comparisons of Global and Local Search Methods to determine the parameters such as maximum number of 250,000 energy evaluations as well as generations of 27,000, followed by mutation and crossover rates of 0.02 and 0.80. The number of docking runs was set to 10. Molecular dynamics study was done to check the stability of the complex. Results: Among all the genus of Zingiberaceae family investigated in this study, Curcuma angustifolia and Hedychium sp. exhibited the highest 537 ± 12.45; 292 ± 9.16 mg gallic acid equivalent/g total polyphenols and 38 ± 1.54; 75 ± 6.75 mg quercetin equivalent/g flavonoids, respectively. Depending on the concentration, the Hedychium sp. extract exerted the highest scavenging activity on DPPH radical (IC50 36.4 μ g/mL). In silico result demonstrated that the synergetic effects of β -phellandrene with other compounds might be responsible for its anticancerous activity. β -phellandrene and farnesene epoxide showed bonding with Leu298, Ala302, Met336, Leu339, Leu343, Phe356, Ala302, Glu 305, Met340, Leu343, Arg346, Phe356, Ile373, Ile376, Leu380, His475, Leu476, and Leu491. Conclusion: Based on the current available literature, this is the first study to understand the interaction of compounds found in the rhizomes of Zingiberaceae family. Abbreviations used: CADD: Computer-aided drug designing; ROS: Reactive oxygen species; ADMET: Absorption, distribution, metabolism, and excretion-toxicity; FeCl3: Ferric chloride; DPPH: 2,2-diphenyl-1-picryl-hydrazyl; NaNO2: Sodium nitrite; TCA: Trichloroacetic acid; K2HPO4: Di-potassium hydrogen phosphate; H2O2: Hydrogen peroxide; KH2PO4: Potassium di-hydrogen phosphate, K2Fe (CN)6: Potassium ferricyanide; KOH: Potassium hydroxide; NaOH: Sodium hydroxide; Na2CO3: Sodium carbonate; CH3COONa: Sodium acetate; AlCl3: Aluminum chloride.
  2,464 175 1
Antioxidant phytochemicals of Opuntia ficus-indica (L.) Mill. cladodes with potential anti-spasmodic activity
Francesco Lanuzza, Francesco Occhiuto, Maria Teresa Monforte, Maria Marcella Tripodo, Valeria D'Angelo, Enza Maria Galati
July-September 2017, 13(51):424-429
DOI:10.4103/pm.pm_495_16  PMID:29142394
Background: Opuntia ficus-indica (OFI) (L.) Mill. (Cactaceae), a plant widespread in dry regions of the world, shows interesting biological activities (cicatrizant, antiulcer, anti-inflammatory, and hypolipidemic) and is widely used in traditional medicine. Objectives: Phytochemical analysis and antispasmodic effect of wild OFI cladodes were carried out. Material and Methods: Polyphenols and Vitamin E occurrence, in antioxidant pool of OFI cladodes, were quantified by high-performance liquid chromatography. The antispasmodic effect of OFI cladodes was assessed in isolated rabbit smooth muscle tissues. The experiments were carried out with preparations of rabbit jejunum and uterus with the spontaneous contractile activity, to evaluate the effect of cumulative concentrations of the extract on basal tone, amplitude, and frequency of contractions. Results: Catechin, quercetin, kaempferol, isorhamnetin and chlorogenic, ferulic, and p-coumaric acid were identified. α -, β -, and γ -tocopherols have been highlighted and α -tocopherol is the major component. OFI cladodes contain significant amount of polyphenols and tocopherols that are effective radical scavengers and inhibited ethanol 1,1-diphenyl-2-picrylhydrazyl formation by 50%. OFI cladodes caused a light inhibition of amplitude and frequency of spontaneous contractions and a marked decrease in muscle basal tone of rabbit jejunum preparations. On spontaneously contracting uterus preparations, the addition of increasing concentrations of cladode extract caused uterine muscle relaxation. Conclusion: The contraction of smooth muscle preparations depends on an increase in cytoplasmic free calcium ion concentration, which activates the contractile elements. The flavonoids may suppress the contractility of smooth myocytes, by an inhibition of availability of Ca2+ for muscle contraction. Abbreviations used: OFI: Opuntia ficus-indica, DPPH: Ethanol 1,1-diphenyl-2-picrylhydrazyl.
  2,488 147 5
Withaferin-A induces apoptosis in osteosarcoma U2OS cell line via generation of ROS and disruption of mitochondrial membrane potential
Hui-Liang Zhang, Hong Zhang
July-September 2017, 13(51):523-527
DOI:10.4103/0973-1296.211042  PMID:28839383
Background: Withaferin-A (WF-A) is a well-known dietary compound isolated from Withania sominifera. It has tremendous pharmacological potential and has been shown to exhibit antiproliferative activity against several types of cancerous cells. Currently, the main focus of anti-cancer therapeutic development is to identify apoptosis inducing drug-like molecules. Osteosarcoma is a rare type of osteocancer, affecting human. The present study therefore focused on the evaluation of antitumor potential of WF-A against several osteosarcoma cell lines. Materials and Methods: MTT assay was used to evaluate WF-A against osteosarcoma cell lines and to calculate the IC50. DAPI staining was used to confirm the apoptosis inducing potential of WF-A. Mitochondrial membrane potential, reactive oxygen species (ROS) assay, and Western blotting were used to confirm the basis of apoptosis. Results: The results revealed that that WF-A exhibited strong antiproliferative activity against all the cells lines, with IC50 ranging from 0.32 to 7.6 μM. The lowest IC50 (0.32 μM) was observed against U2OS cell line and therefore it was selected for further analysis. DAPI staining indicated that WF-A exhibited antiproliferative activity via induction of apoptosis. Moreover, WF-A induced ROS-mediated reduction in mitochondrial membrane potential ΔΨm) in a dose-dependent manner and activation of caspase-3 in osteosarcoma cells. Conclusion: We propose that WF-A may prove a potent therapeutic agent for inducing apoptosis in osteosarcoma cell lines via generation of ROS and disruption of mitochondrial membrane potential. Abbreviations used: WA: Withaferin A; ROS: Reactive oxygen species; OS: Osteosarcoma; MMP: Mitochondrial membrane potential.
  2,515 104 3
In Silico and In Vitro anticancer activity of isolated novel marker compound from chemically modified bioactive fraction from Curcuma longa (NCCL)
Arshi Naqvi, Richa Malasoni, Swati Gupta, Akansha Srivastava, Rishi R Pandey, Anil Kumar Dwivedi
July-September 2017, 13(51):640-644
DOI:10.4103/pm.pm_23_17  PMID:29142426
Background: Turmeric (Curcuma longa) is reported to possess wide array of biological activities. Herbal Medicament (HM) is a standardized hexane-soluble fraction of C. longa and is well known for its neuroprotective effect. Objective: In this study, we attempted to synthesize a novel chemically modified bioactive fraction from HM (NCCL) along with isolation and characterization of a novel marker compound (I). Materials and Methods: NCCL was prepared from HM. The chemical structure of the marker compound isolated from NCCL was determined from 1D/2D nuclear magnetic resonance, mass spectroscopy, and Fourier transform infrared. The compound so isolated was subjected to in silico and in vitro screenings to test its inhibitory effect on estrogen receptors. Results: Molecular docking studies revealed that the binding poses of the compound I was energetically favorable. Among NCCL and compound I taken for in vitro studies, NCCL had exhibited good anti-cancer activity over compound I against MCF-7, MDA-MB-231, DU-145, and PC-3 cells. Conclusion: This is the first study about the synthesis of a chemically modified bioactive fraction which used a standardized extract since the preparation of the HM. It may be concluded that NCCL fraction having residual components induce more cell death than compound I alone. Thus, NCCL may be used as a potent therapeutic drug.
  2,463 139 1
Flavones and lignans from the stems of Wikstroemia scytophylla diels
Jie-Ping Lei, Juan-Juan Yuan, Sheng-Hao Pi, Rui Wang, Rui Tan, Chao-Ying Ma, Tao Zhang, He-Zhong Jiang
July-September 2017, 13(51):488-491
DOI:10.4103/pm.pm_275_16  PMID:28839377
Background: The genus Wikstroemia has about 70 species, but only a limited number of species have been studied chemically. Wikstroemia indica has long been used as a traditional crude drug in China. However, there is no report about the bioactivity of Wikstroemia scytophylla. Objective: This paper reports the chemical investigation and biological evaluation of the W. scytophylla. Materials and Methods: The EtOAc extraction of W. scytophylla was isolated using chromatographic methods, and the compounds were analyzed by spectroscopic methods. The in vitro antitumor activities against five human cancer cell lines were performed according to the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Results: The chemical investigation of the stems of W. scytophylla resulted in the isolation of 12 compounds mainly including one biflavone (1), five flavones (2-6) compounds, and six lignans (7-12), in which compound 8 was a new natural product. Compounds 1 and 7-12 were evaluated for their antitumor activities while these compounds showed weak cytotoxicity with the half maximal inhibitory (IC50) values more than 40 μM. Conclusion: All of these compounds were isolated from this plant for the first time, and compounds 2-12 were first reported from genus Wikstroemia, in which compound 8 was a new natural product. Compounds 1 and 7-12 exhibited weak antitumor activities (IC50>40 μM). The chemotaxonomic significance of all the isolations was summarized. Abbreviations used: MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; IC50: Half maximal inhibitory; HL-60: Human leukemia cell line; SMMC-7721: Human hepatocellular carcinoma cell line; A549: Human lung tumor cell line; MCF-7: Human breast cancer cell line; SW480: Human colon cancer cell line; MS: Mass spectrometry; NMR: Nuclear Magnetic Resonance.
  2,477 105 2
Quality evaluation of raw moutan cortex using the ahp and gray correlation-TOPSIS Method
Sujuan Zhou, Bo Liu, Jiang Meng
July-September 2017, 13(51):528-533
DOI:10.4103/0973-1296.211029  PMID:28839384
Background: Raw Moutan cortex (RMC) is an important Chinese herbal medicine. Comprehensive and objective quality evaluation of Chinese herbal medicine has been one of the most important issues in the modern herbs development. Objective: To evaluate and compare the quality of RMC using the weighted gray correlation- Technique for Order Preference by Similarity to an Ideal Solution (TOPSIS) method. Materials and Methods: The percentage composition of gallic acid, catechin, oxypaeoniflorin, paeoniflorin, quercetin, benzoylpaeoniflorin, paeonol in different batches of RMC was determined, and then adopting MATLAB programming to construct the gray correlation-TOPSIS assessment model for quality evaluation of RMC. Results: The quality evaluation results of model evaluation and objective evaluation were consistent, reliable, and stable. Conclusion: The model of gray correlation-TOPSIS can be well applied to the quality evaluation of traditional Chinese medicine with multiple components and has broad prospect in application.
  2,442 109 2
Antioxidant and hepatoprotective potential of Swaras and Hima extracts of Tinospora cordifolia and Boerhavia diffusa in Swiss albino mice
Amit Kaushik, Athar Husain, Harshika Awasthi, Dewasya Pratap Singh, Raziuddin Khan, Dayanandan Mani
July-September 2017, 13(51):658-662
DOI:10.4103/pm.pm_448_16  PMID:29142429
Background: In Ayurveda, five basic extraction procedures are mentioned in order of their decreasing potency. Swaras is considered as the most potent followed by, kalka, kwatha, fanta and hima. Objective: Present study was carried out to investigate the antioxidant and hepatoprotective potential of swaras and hima extracts of T.cordifolia and B. diffusa. Materials and Methods: Swaras and hima extracts of T. cordifolia and B. diffusa were prepared. Phytochemical screening and in vitro antioxidant activities was carried out using standard methods. Hepatoprotective efficacy of extracts were carried out in Swiss albino mice using paracetamol induced hepatotoxicity. Animals were administered with swaras and hima extracts of both plants at 200 mg/kg BW dose for 7 days and on 8th day hepatotoxicity was induced by intraperitoneal injection of paracetamol at 500 mg/kg BW. The degree of liver protection was determined by measuring the levels of liver enzymes followed by histopathology. Results and Discussion: The results of phytochemical, antioxidant and hepatoprotective activities showed that there were no significant difference between swaras and hima extracts. Both the extract of T. cordifolia were equally potent in reducing SGOT (P < 0.01) and ALP level (P < 0.001). Similar effects were observed with the Swaras and hima extracts of B. diffusa. Both the extracts reduced SGOT and ALP (P < 0.01). Histopathological findings among all the extracts were also more or less similar in lowering the paracetamol mitigated necrosis. Conclusion: The present study suggested that T. cordifolia and B. diffusa possess potential hepatoprotective activity irrespective of the extraction procedure. Abbreviations used: TC swaras: T. cordifolia swaras; TC hima: T. cordifolia hima; BD swaras: B. diffusa swaras; BD hima: B. diffusa hima; BW: Body weight; LDL: Low-density lipoprotein; HDL: High-density lipoprotein; SGOT: Serum glutamate oxaloacetate transminase; SGPT: Serum glutamate pyruvate transminase; ALP: Alkaline phosphatase; I.P: Intraperitoneal; TAC: Total antioxidant capacity; DPPH: 2,2-diphenyl-1-picrylhydrazyl; TCA: Trichloro acetic acid; NO: Nitric oxide; TPC: Total phenolic content; NAPQI: N-acetyl-p-benzoquinone imine; PCM: Paracetamol.
  2,411 135 7
Biological activity and isolation of compounds from stem bark of Plumeria acutifolia
Ghadah A Alhozaimy, Ebtesam Saad Al-Sheddi, Taghreed A Ibrahim
July-September 2017, 13(51):505-511
DOI:10.4103/pm.pm_22_17  PMID:29142406
Background: Plumeria acutifolia (Apocynaceae) is an ornamental plant, used in the traditional medicine and known to have a variety of constituents as alkaloids, flavonoids, and iridoids. Extracts of this plant were proved to have antimicrobial and anticancer activities. Objectives: This research was conducted for the evaluation of the biological activities of P. acutifolia stem bark and isolation and structural elucidation of various chemical compounds from the biologically active fractions. Materials and Methods: Methanol extract of stem bark of P. acutifolia was successively fractionated with petroleum ether, dichloromethane, ethyl acetate, and n-butanol. The fractions were evaluated for their antimicrobial, cytotoxic, and antioxidant activities. Fractions with promising biological activities were subjected to chromatographic techniques for the isolation of compounds, followed by structural elucidation using several spectroscopic techniques. Results: P. acutifolia stem bark showed a significant antimicrobial activity, where the ethyl acetate fraction was active against Syncephalastrum racemosum (7.81 μ g/ml) and Escherichia coli (3.9 μ g/ml). The cytotoxic activity against HEPG-2, HCT-116, and MCF-7 cell lines was highest in the petroleum ether fraction, using concentrations of 1, 2.5, 5, and 10 μ l/ml. The antioxidant activity was concentration dependent; ethyl acetate fraction showed the most predominant effect, with an IC50of 197.1 μ g/ml. Five compounds were identified as narcissin (1), quercitrin (2), sweroside (3), gaertneroside (4), and plumieride (5). Conclusion: P. acutifolia was proved to have significant antimicrobial, cytotoxic, and antioxidant activities; the isolated compounds were flavonoids, iridoids, and secoiridoid, some of which were reported for the first time in genus Plumeria and/or family Apocynaceae. Abbreviations used: mp: Melting point, NMR: Nuclear magnetic resonance, s: Singlet, d: Double, t: Triplet, q: Quartet, dd: Double-double, m: Multiplet, br: Broad.
  2,382 162 1
Protective effect of Pluchea lanceolata against aluminum chloride-induced neurotoxicity in Swiss albino mice
Ravi Mundugaru, Senthilkumar Sivanesan, Padmaja Udaykumar, Niranjan Rao, Naveen Chandra
July-September 2017, 13(51):567-572
DOI:10.4103/pm.pm_124_17  PMID:29142416
Background: Aluminum chloride (AlCl3) is a known potent environmental neurotoxin causing progressive neurodegenerative changes in the brain. The herb Pluchea lanceolata is commonly known as “Rasana” and used as a nerve tonic in neuroinflammatory conditions in Indian system of medicine. Objective: To evaluate the neuroprotective activity of hydroalcoholic extract of P. lanceolata in chronic AlCl3-induced neurotoxicity in Swiss albino mice. Materials and Methods: Albino mice were categorized into four different groups; Group 1served as vehicle control, Group 2 mice were administered with AlCl3, 40 mg/kg body weight by intraperitoneal route for 45 consecutive days. Groups 3 and 4 mice were administered with AlCl3, 40 mg/kg body weight intraperitoneal for 45 consecutive days along with hydroalcoholic extract of P. lanceolata at 200 and 400 mg/kg body weight. Results: Chronic administration of AlCl3resulted in behavioral deficits, triggered lipid peroxidation, increased acetylcholinesterase (AChE) activity, and histological alterations. Co-administration of hydroalcoholic extract of P. lanceolata attenuated many of the AlCl3-induced alterations such as behavioral, lipid peroxidation, AChE, and histological changes of brain tissue. Conclusion: The results of the present study have demonstrated the protective role of hydroalcoholic extract of P. lanceolata against AlCl3-induced neurotoxicity in Swiss albino mice. The neuroprotective efficacy of P. lanceolata can help reduce the symptoms caused by toxic protein aggregates in several degenerative diseases. Abbreviations used: HAPL: Hydro alcoholic extract of Pluchea lanceolata; CAT: Catalase; GSH-Px: Glutathione peroxidase; SOD: Superoxide dismutase; TBARS: Thio-barbituric acid reactive substances; MDA: Malondialdehyde; AChE: Acetylcholine esterase; AOT: Acute oral toxicity; CNS: Central nervous system; H2O2: Hydrogen peroxide; ML: molecular layer; GL: granular layer; MC: microcytic changes; BV: blood vessels; DG: dentate gyrus; PC: pyramidal cells; LD: Lethal dose; ANOVA: Analysis of variance; SEM: Standard error of mean; PCL: Pyramidal cell layer; OCL: Outer granular layer; BV: blood vessels; PM: Pia mater.
  2,424 109 -
Rapid detection of volatile oil in Mentha haplocalyx by near-infrared spectroscopy and chemometrics
Hui Yan, Cheng Guo, Yang Shao, Zhen Ouyang
July-September 2017, 13(51):439-445
DOI:10.4103/0973-1296.211026  PMID:28839369
Near-infrared spectroscopy combined with partial least squares regression (PLSR) and support vector machine (SVM) was applied for the rapid determination of chemical component of volatile oil content in Mentha haplocalyx. The effects of data pre-processing methods on the accuracy of the PLSR calibration models were investigated. The performance of the final model was evaluated according to the correlation coefficient (R) and root mean square error of prediction (RMSEP). For PLSR model, the best preprocessing method combination was first-order derivative, standard normal variate transformation (SNV), and mean centering, which had Rc2 of 0.8805, Rp2 of 0.8719, RMSEC of 0.091, and RMSEP of 0.097, respectively. The wave number variables linking to volatile oil are from 5500 to 4000 cm−1 by analyzing the loading weights and variable importance in projection (VIP) scores. For SVM model, six LVs (less than seven LVs in PLSR model) were adopted in model, and the result was better than PLSR model. The Rc2 and Rp2 were 0.9232 and 0.9202, respectively, with RMSEC and RMSEP of 0.084 and 0.082, respectively, which indicated that the predicted values were accurate and reliable. This work demonstrated that near infrared reflectance spectroscopy with chemometrics could be used to rapidly detect the main content volatile oil in M. haplocalyx. Abbreviations used: 1st der: First-order derivative; 2nd der: Second-order derivative; LOO: Leave-one-out; LVs: Latent variables; MC: Mean centering, NIR: Near-infrared; NIRS: Near infrared spectroscopy; PCR: Principal component regression, PLSR: Partial least squares regression; RBF: Radial basis function; RMSEC: Root mean square error of cross validation, RMSEC: Root mean square error of calibration; RMSEP: Root mean square error of prediction; SNV: Standard normal variate transformation; SVM: Support vector machine; VIP: Variable Importance in projection
  2,380 107 3
Berberine depresses contraction of smooth muscle via inhibiting myosin light-chain kinase
Zhili Xu, Mingbo Zhang, Deqiang Dou, Xiaojun Tao, Tingguo Kang
July-September 2017, 13(51):454-458
DOI:10.4103/pm.pm_205_16  PMID:28839371
Background: Berberine is a natural isoquinoline alkaloid possessing various pharmacological effects, particularly apparent in the treatment of diarrhea, but the underlying mechanism remains unclear. Smooth muscle myosin light-chain kinase (MLCK) plays a crucial role in the smooth muscle relaxation-contraction events, and it is well known that berberine can effectively depress the contraction of smooth muscle. Hence, whether berberine could inhibit MLCK and then depress the smooth muscle contractility might be researched. Objective: The purpose of this study is to investigate the effects of berberine on MLCK. Based on this, the contractility of gastro-intestine, catalysis activity of MLCK, and molecular docking are going to be evaluated. Materials and Methods: The experiment of smooth muscle contraction was directly monitored the contractions of the isolated gastrointestine by frequency and amplitude at different concentration of berberine. The effects of berberine on MLCK were measured in the presence of Ca2+-calmodulin, using the activities of 20 kDa myosin light chain (MLC20) phosphorylation, and myosin Mg2+-ATPase induced by MLCK. The docking study was conducted with expert software in the meantime. Results: The phosphorylation of myosin and the Mg2+-ATPase activity is reduced in the presence of berberine. Moreover, berberine could inhibit the contractibility of isolated gastric intestine smooth muscle. Berberine could bind to the ATP binding site of MLCK through hydrophobic effect and hydrogen bonding according to the docking study. Conclusion: The present work gives a deep insight into the molecular mechanism for the treatment of diarrhea with berberine, i.e., berberine could suppress the contractility of smooth muscle through binding to MLCK and depressing the catalysis activity of MLCK. Abbreviations used: MLCK: Myosin light chain kinase; MLC20: 20 KDa regulating myosin light chain; CaM: Calmodulin.
  2,339 117 6
Determination and tissue distribution comparisons of atractylodin after oral administration of crude and processed atractylodes rhizome
Yang-Zhi Liu, Yu-Qiang Liu, Rui Jia, Jun Li, Xiao-Wen Chang, Chen-Xi Xu, Qian Cai
July-September 2017, 13(51):413-417
DOI:10.4103/pm.pm_394_16  PMID:28839365
Background: Atractylodis rhizoma is one of the most often used drugs in traditional Chinese medicine. Stir frying with wheat bran is the most common processing method. To clarify the principle of processing, an experiment was carried out to compare the tissue distribution of typical constituent after oral administration of raw A. rhizoma and processed ones. Objective: To compare the tissues distribution of atractylodin after oral administration of raw and processed A. rhizoma and clarify the processing principle of A. rhizoma. Materials and Methods: High-performance liquid chromatogram with ultraviolet detection was developed and validated for the determination of atractylodin in rat tissues. Results: The atractylodin in raw and processed A. rhizoma was distributed in all tissues involved in this study. Conclusions: The concentration of atractylodin in it is the highest in the stomach and small intestine. Abbreviations used: IS: Internal standard substance; A. Rhizoma: Atractylodis rhizoma; RSD: Relative standard deviation; HPLC: High performance liquid chromatography.
  2,349 99 2
Characterization of the phenolic compound, gallic acid from Sansevieria roxburghiana schult and schult. f. rhizomes and antioxidant and cytotoxic activities evaluation
Rajalekshmi Maheshwari, Chandrashekara Shastry Shreedhara, Picheswara Rao Polu, Renuka Suresh Managuli, Seena Kanniparambil Xavier, Richard Lobo, Manjunath Setty, Srinivas Mutalik
July-September 2017, 13(51):693-699
DOI:10.4103/pm.pm_497_16  PMID:29142435
Background: Sansevieria roxburghiana Schult. and Schult. f. (Asparagaceae) grows in India, Indonesia, Sri Lanka, and tropical Africa. Even though the plant has been traditionally used for the treatment of many ailments, the antioxidant and antiproliferative activities of S. roxburghiana methanol extract and its fractions have not yet been explored. Materials and Methods: Quantitative estimation of phenols and different antioxidant assays were performed using standard methods. Anti-proliferative effect of the extract and fractions were evaluated in HCT-116, HeLa, MCF-7, HepG2, and A-549 cancer cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and sulforhodamine B (SRB) assay methods. High-performance liquid chromatography (HPLC) and high-performance thin layer chromatography (HPTLC) fingerprint profiling were carried out for extract and different fractions. Results: Significant antioxidant and anti-proliferate activity were detected in ethyl acetate fraction. Ethyl acetate fraction showed prominent scavenging activity in 1,1-diphenyl-2-picrylhydrazyl, 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, and nitric oxide antioxidant assays with an concentration yielding 50% inhibition (IC50) 15.33 ± 1.45, 45.3 ± 1.93 and 48.43 ± 0.46 μ g/ml, respectively. Cytotoxicity of ethyl acetate fraction was the highest among other fractions against HCT-116, HeLa, and MCF-7cancer cell lines with IC50values 16.55 ± 1.28, 12.38 ± 1.36, and 8.03 ± 1.9 μ g/ml, respectively, by MTT assay and 15.57 ± 0.70, 13.19 ± 0.49, and 10.34 ± 0.9 μ g/ml, respectively, by SRB assay. The presence of gallic acid in the ethyl acetate fraction of S. roxburghiana rhizomes was confirmed by HPLC and HPTLC analysis. Conclusion: Results suggested that ethyl acetate fraction exhibited effective antioxidant and antiproliferative activities. The phenolic compounds identified in ethyl acetate fraction could be responsible for the activities. Abbreviations used: %: Percent, °C: Celsius, μ g: Microgram, μ l-Microlitre, ANOVA: Analysis of variance, DMSO: Dimethyl sulfoxide, g: Grams, IC50: Concentration yielding 50% inhibition, Kg: Kilogram, mg: Milligram, min: Minutes, ml: Milliliter, HPLC: High-performance liquid chromatography, HPTLC: High-performance thin layer chromatography, DPPH: 1,1-diphenyl-2-picrylhydrazyl, ABTS: 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, GAE: Gallic acid equivalents, SRME: Methanol extract of S. roxburghiana, ROS: Reactive oxygen species, SRPE: Petroleum ether fraction of S. roxburghiana, SREA: Ethyl acetate fraction of S. roxburghiana, SRAQ: Aqueous fraction of S. roxburghiana, DMEM: Dulbecco's Minimum Essential Medium, FBS: Fetal bovine serum, OD: Optical density, TPC: Total phenolic content, SRBU: Butanol fraction of S. roxburghiana.
  2,303 114 2
Biological effect of Cynara cardunculus on kidney status of hypercholesterolemic rats
Abdullah Glil Alkushi
July-September 2017, 13(51):430-436
DOI:10.4103/pm.pm_14_17  PMID:29142395
Context: Cynara cardunculus or artichoke thistle belongs to the sunflower family and has a variety of cultivable forms. Historically, it was cultivated as a vegetable, but more recently, it is being used in cheese and biofuel preparation. Artichoke leaf extracts are also known for its medicinal purposes, particularly in reducing the elevated cholesterol levels in blood. Hypercholesterolemia (HC) is also associated with other complications such as impaired renal function and diabetes mellitus. A remedy without major side effects for HC and its associated complications is highly desirable. Aims: We explored the effect of artichoke on the kidneys of hypercholesterolemic adult male Sprague–Dawley albino rats. Subjects and Methods: Oral administration of 200 mg/kg and 400 mg/kg body weight (b.wt.) of C. cardunculus leaf extract (CCL) and C. cardunculus pulp extract (CCP) was made to male Sprague–Dawley albino hypercholesterolemic rats and investigated the levels of glucose, creatinine, uric acid, and urea in their blood. Results: We observed that both CCL and CCP significantly reduced the creatinine and uric acid levels in the blood in a dose-dependent manner (P < 0.05). Both CCL and CCP significantly reduced the blood glucose levels (P < 0.05). Further, the histopathological investigation of the kidney sections showed that CCL treatment resolved HC-associated kidney damage. Conclusion: CCL not only has cholesterol-reducing capacity but also reduces the blood glucose levels and repairs the impaired kidney functions and damages. These findings are significant particularly because HC results in further complications such as diabetes and kidney damage, both of which can be treated effectively with artichoke. Abbreviations used: HC: Hypercholesterolemia, WHO: World Health Organization, BAS: Bile acid sequestrant, PCSK9: Proprotein convertase subtilisin kexin type 9, ALE: Artichoke leaf extract, CCL: Cynara cardunculus leaf extract, CCP: Cynara cardunculus pulp extract, BWG%: Body weight gain%, FER: Food-efficiency ratio.
  2,300 104 1
Docking-based screening of Ficus religiosa phytochemicals as inhibitors of human histamine H2 receptor
Amit Chaudhary, Birendra Singh Yadav, Swati Singh, Pramod Kumar Maurya, Alok Mishra, Shweta Srivastva, Pritish Kumar Varadwaj, Nand Kumar Singh, Ashutosh Mani
July-September 2017, 13(51):706-714
DOI:10.4103/pm.pm_49_17  PMID:29142437
Background: Ficus religiosa L. is generally known as Peepal and belongs to family Moraceae. The tree is a source of many compounds having high medicinal value. In gastrointestinal tract, histamine H2 receptors have key role in histamine-stimulated gastric acid secretion. Their over stimulation causes its excessive production which is responsible for gastric ulcer. Objective: This study aims to screen the range of phytochemicals present in F. religiosa for binding with human histamine H2 and identify therapeutics for a gastric ulcer from the plant. Materials and Methods: In this work, a 3D-structure of human histamine H2 receptor was modeled by using homology modeling and the predicted model was validated using PROCHECK. Docking studies were also performed to assess binding affinities between modeled receptor and 34 compounds. Molecular dynamics simulations were done to identify most stable receptor-ligand complexes. Absorption, distribution, metabolism, excretion, and screening was done to evaluate pharmacokinetic properties of compounds. Results: The results suggest that seven ligands, namely, germacrene, bergaptol, lanosterol, Ergost-5-en-3beta-ol, α -amyrin acetate, bergapten, and γ -cadinene showed better binding affinities. Conclusion: Among seven phytochemicals, lanosterol and α -amyrin acetate were found to have greater stability during simulation studies. These two compounds may be a suitable therapeutic agent against histamine H2 receptor. Abbreviations used: ADMET: Absorption, distribution, metabolism, excretion and toxicity, DOPE: Discrete Optimized Potential Energy, OPLS: Optimized potential for liquid simulations, RMSD: Root-mean-square deviation, HOA: Human oral absorption, MW: Molecular weight, SP: Standard-precision, XP: Extra-precision, GPCRs: G protein-coupled receptors, SASA: Solvent accessible surface area, Rg: Radius of gyration, NHB: Number of hydrogen bond
  2,250 153 1
A comprehensive study on fast dispersible and slow-releasing characteristic of orange peel pectin in relation to established synthetic polymer
Pranati Srivastava, Mahendra Singh, Shilpi Bhargava
July-September 2017, 13(51):401-404
DOI:10.4103/pm.pm_193_17  PMID:29142390
Introduction: In the present work, the method to extract, isolate, and characterize orange peel pectin using soxhlation, and thereafter, the use of this polymer–polymer in the formulation of fast dispersable and slow-releasing tablet has been studied. Thereafter, the evaluation and comparison of fast dispersible/slow-releasing tablets using orange peel pectin versus prepared using sodium starch glycolate (SSG) were carried out. Materials and Methods: In the present investigation, extraction methodology was employed for isolation of pectin from orange peels. Four different batches with each polymer were prepared with varying concentration of superdisintegrant and bulking agent using diclofenac sodium as model drug. Diclofenac sodium stands as easily available, cheap, and good candidate to demonstrate disintegrant property. The formulation involved wet granulation method for the preparation of tablets of each batch. The tablets were evaluated for hardness, friability, thickness, wetting time, deaggregation time, and in vitro release characteristic data. Results: It was observed that parameters for batch O2* were comparable with that of synthetic superdisintegrant. This batch gave around 92.12% drug release in period of 90 min. The study showed that orange peel pectin could be a potential candidate for formulation of orodispersible dosage forms in competence to SSG, which is established superdisintegrant. Conclusion: The results led to the conclusion that the use of natural polymers in formulation of pharmaceutical dosage form can be put into practice on industrial scale meeting the similar requirements as done by synthetic polymers. Abbreviations used: O1–O2: Batches Containing Orange peel pectin, S1–S2: Batches containing SSG, SSG: Sodium starch glycolate, NDDS: Novel drug delivery system.
  2,291 96 -
Cytotoxic, antimitotic, and antiproliferation studies on Rasam: A South Indian traditional functional food
Agilandeswari Devarajan, MK Mohan Maruga Raja
July-September 2017, 13(51):452-457
DOI:10.4103/pm.pm_138_17  PMID:29142398
Background: Rasam is a traditional South Indian food, prepared using tamarind juice as a base, with a variety of spices. Rasam, with all its ingredients medicinally claimed for various ailments, is a functional food. Systematic consumption of traditional functional food provides an excellent preventive measure to ward off many diseases. Objective: To study rasam for cytotoxic, antimitotic, and antiproliferation potential beyond its culinary and nutritional effect. Materials and Methods: Brine shrimp lethality assay, onion root tip inhibition assay, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay in Calu-6, HeLa, MCF-7 cell lines for four stage-wise samples in the preparation of rasam (RS1, RS2, RS3, and RS4) were studied. Results: RS4, the end product of rasam showed high lethality with an LC50value of 38.7 μ L/mL. It showed maximum antimitotic activity in a dose-dependent manner compared to other samples with an IC50value of 189.86 μ L/mL. RS4 also showed an IC50value of 350.22 and 410.15 μ L/mL in MCF-7 and Calu-6 cell lines, respectively. Conclusion: From this study, we suggest that rasam is a classic example of traditional functional food and it can treat breast and lung cancer on chronic use. Abbreviations used: SS 316: Stainless Steel 316 grade; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; DMEM: Dulbecco's modified Eagle medium; FBS: Fetal bovine serum media; TPVG: Trypsin phosphate versene glucose; EDTA: Ethylene diamine tetra acetic acid; PBS: Phosphate buffered saline; DMSO: Dimethyl sulfoxide.
  2,292 92 2
Inhibition of glycation-induced cytotoxicity, protein glycation, and activity of proteolytic enzymes by extract from Perovskia atriplicifolia Roots
Mehran Miroliaei, Akram Aminjafari, Sylwester Ślusarczyk, Izabela Nawrot-Hadzik, Mehdi Rahimmalek, Adam Matkowski
July-September 2017, 13(51):676-683
DOI:10.4103/pm.pm_559_16  PMID:29142432
Background: Protein glycation and glycotoxicity belong to the main oxidative-stress related complications in diabetes. Perovskia species are used in Asian folk medicine as antidiabetic herbs. Objective: The aim of this study was to verify the ability of the methanolic extract from Perovskia atriplicifolia Benth. roots to diminish glycation of albumin and to prevent cell damage in vitro. Furthermore, we tested the extract for in vitro antioxidant activity and inhibition of elastase and collagenase. Material and Methods: The aqueous methanol extract was analyzed by UHPLC-MS for the content of polyphenols and terpenoids. The prevention of glycated albumin-induced cell damage was tested in four mammalian cell lines (peripheral blood mononuclear cells, human embryonic kidney cells – HEK293, normal human fibroblasts, and Chinese hamster ovary cells) with the 5-(3-carboxymethoxyphenyl)-2-(4,5-dimethylthiazoly)-3-(4-sulfophenyl) tetrazolium assay. Results: Glycated albumin is significantly more toxic than native human serum albumin (LC50from 35.00 to 48.34 μ g/mL vs. 5.47–9.10 μ g/mL, respectively). The extract, rich in rosmarinic acid (344.27 mg/g dry mass), mitigated the glycated albumin toxicity, and increased glycated albumin-treated cell survival by more than 50%. The inhibition of advanced glycation endproduct formation was confirmed by monitoring conformational changes. The free radical scavenging activity was higher than Trolox and metal reducing power was one-third to half that of ascorbic acid. The activity of elastase and collagenase was inhibited by 54.75% ± 6.87% and 60.03% ± 7.22%, respectively. Conclusions: The results confirm antiglycative and antiglycotoxic potential of Perovskia root and its traditional antidiabetic use. The high activity can be attributed to rosmarinic acid abundance. Abbreviations used: AGE: advanced glycation end-products; DPPH: 2,2-diphenyl-1-picrylhydrazyl; HSA: human serum albumin.
  2,207 168 4
Methanolic extract of Costus pictus D. DON induces cytotoxicity in liver hepatocellular carcinoma cells mediated by histone deacetylase inhibition
PV Neethu, K Suthindhiran, MA Jayasri
July-September 2017, 13(51):533-538
DOI:10.4103/pm.pm_524_16  PMID:29142410
Background: Leaves of Costus pictus D. Don, (insulin plant) are used as dietary supplement for the treatment of diabetes. Objective: The antidiabetic activity of this plant is well documented, but its activity on different cell types and mechanism remains unknown. Thus, the present study evaluates the cytotoxicity of C. pictus methanolic extract (CPME) against various cancer and normal cells. Materials and Methods: Dried leaves of C. pictus were extracted using methanol and were subjected to histone deacetylase (HDAC) inhibition and toxicity studies. Results: The CPME displayed a selective toxicity toward tested cancer cells in a dose- and time-dependent manner. CPME exhibited significant cytotoxicity on Liver hepatocellular carcinoma cells (Hep G2) (half maximal inhibitory concentration IC50 = 6.7 μ g/ml). Since CPME demonstrates both antidiabetic, anticancer activity, and HDAC enzyme play a detrimental role in both the complications, we have evaluated the CPME-induced HDAC regulation on Hep G2 cell lines. CPME showed a notable HDAC inhibition (55%). Furthermore, CPME did not show any genotoxicity or membrane instability at the tested concentrations. Conclusion: CPME demonstrates selective cytotoxicity toward tumor cells at a lower concentration through HDAC inhibition. Abbreviations used: A549: Human lung carcinoma cells, CPME: Costus pictus methanolic extract, DMEM: Dulbecco's modified eagle's medium, DMSO: Dimethyl sulfoxide, ELISA: Enzyme-linked immunosorbent assay, 5-FU: 5-Fluorouracil, Hep G2: Liver hepatocellular carcinoma cells, HEK-293: Human embryonic kidney cells, Hela: Human cervical carcinoma cells, HT-29: Human colorectal adenocarcinoma cells, HDAC: Histone deacetylase, IC50: Half maximal inhibitory concentration, MCF-7: Human breast adenocarcinoma cells, MDA-MB-435S: Human breast cancer cells, MTT: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide, NFF: Neonatal foreskin fibroblasts, PHA: Phytohemagglutinin, PBS: Phosphate buffer saline, RPMI-1640: Roswell Park Memorial Institute Medium.
  2,254 114 -
Antiurolithiatic potential of neeri against calcium-oxalate stones by crystallization inhibition, free radicals scavenging, and NRK-52E cell protection from oxalate injury
Parveen Kumar Goyal, Santosh Kumar Verma, Anil Kumar Sharma
July-September 2017, 13(51):549-554
DOI:10.4103/pm.pm_551_16  PMID:29142413
Background: Neeri is a well-established polyherbal formulation prescribed for renal stones by the physicians but has not been experimentally evaluated for its antiurolithiatic potential using cell-lines. Objective: This study is aimed to scientifically substantiate the antiurolithiatic effect of Neeri extract (NRE) through calcium oxalate (CaOx) crystallization inhibition, scavenging of free radicals, and protection of renal tubular epithelial NRK-52E cells from oxalate-induced injury. Materials and Methods: The crystallization inhibition was studied by turbidimetric assay while the free radical scavenging potential was determined for superoxide and nitric oxide (NO) radicals. The cytoprotective effect against oxalate-induced injury was assessed by estimating lactate dehydrogenase (LDH) leakage and determining cell viability using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Results: NRE significantly inhibited the CaOx crystallization in a concentration-dependent manner and also scavenged superoxide (IC50 302.88 μ g/ml) and NO (IC50 300.45 μ g/ml) free radicals. It did not show any significant cytotoxicity for NRK-52E cells till the highest dose (500 μ g/ml) and found to be safe. When NRK-52E cells, injured by exposing to oxalate crystals for 24 h, were treated with NRE, it appreciably prevented the cell injury in a dose-dependent manner. It significantly decreased the elevated LDH leakage toward normal range and improved renal cell viability (82.37% ± 0.87%), hence, prevented growth and retention of crystals. Conclusion: The experimental findings concluded that Neeri is a potent antiurolithiatic formulation that inhibited CaOx crystallization and prevented tubular retention of crystals by protecting the renal cells against oxalate-induced injury as well as reducing the oxidative stress by scavenging free radicals. Abbreviations used: Ac: Absorbance of control, At: Absorbance of test, ANOVA: Analysis of variance, CaOx: Calcium oxalate, DMEM: Dulbecco's Modified Eagle's Medium, DMSO: Dimethyl sulfoxide, EDTA: Ethylenediaminetetraacetic acid, FBS: Fetal bovine serum, INT: Iodonitrotetrazolium, LDH: Lactate dehydrogenase, M: Molar, ml: Milliliter, mM: Millimolar, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, NAD: Nicotinamide adenine dinucleotide, NADPH: Nicotinamide adenine dinucleotide phosphate, NBT: Nitro blue tetrazolium, nm: Nanometer, NO: Nitric oxide, NRE: Neeri extract, PMS: Phenazine methosulfate, ROS: Reactive oxygen species, Sc: Slope of the graph of control, SEM: Standard error of mean, Si: Slope of the graph with inhibitor, U/I: International unit, μg: Microgram, μl: Microliter.
  2,217 121 -
Influence of boiling duration of GCSB-5 on index compound content and antioxidative and anti-inflammatory activity
In-Hee Lee, Hwa-Jin Chung, Joon-Shik Shin, In-Hyuk Ha, Me-Riong Kim, Wonil Koh, Jinho Lee
July-September 2017, 13(51):418-424
DOI:10.4103/pm.pm_425_16  PMID:28839366
Background: GCSB-5, an herbal drug composition with an anti-inflammatory effect, is prepared by boiling, which is the most common herbal extraction method in traditional Korean medicine. Several parameters are involved in the process, i.e., extractant type, herb-to-extractant ratio, extraction temperature and pressure, and total boiling time. Objectives: The aim of this study was to examine the influence of boiling time on index compound amount and the antioxidative and anti-inflammatory activities of GCSB-5. Materials and Methods: Different samples of GCSB-5 were obtained by decocting for 30, 60, 90, 120, 150, and 240 min. Each sample was tested for hydrogen ion concentration (pH), total soluble solid content (TSSC), marker compound profiles, and antioxidative and anti-inflammatory activity. Results: pH was found to decrease while TSSC increased with extended decoction. Marker compound contents for GCSB-5 (acanthoside D for Acanthopanax sessiliflorus Seem, 20-hydroxyecdysone for Achyranthes japonica Nakai, and pinoresinol diglucoside for Eucommia ulmoides Oliver) remained relatively constant regardless of the length of boiling. Total D-glucose amount increased with longer boiling. The antioxidative and anti-inflammatory potentials of GCSB-5 were not substantially affected by decoction duration. Conclusion: Biological characteristics and marker compound content of GCSB-5 were not altered significantly in prolonged boiling. Abbreviations used: CAM: Complementary and alternative medicine; KIOM: Korea Institute of Oriental Medicine; KMD: Korean medicine doctor; TSSC: Total soluble solid content; pH: Hydrogen ion concentration; HPLC: High-performance liquid chromatography; NO: Nitric oxide; NO2: Nitric dioxide; LPS: Lipopolysaccharide; DMSO: Dimethyl sulfoxide.
  2,226 99 2
Chemical composition, antimicrobial and antitumor potentiality of essential oil of Ferula tingitana L. apiaceae grow in Libya
Waleed Elghwaji, Abeer Mohamed El-Sayed, Kadriya S El-Deeb, Aly M ElSayed
July-September 2017, 13(51):446-451
DOI:10.4103/pm.pm_323_15  PMID:29142397
Background: Ferula tingitana L. (Apiaceae) has been considered to have abortive and menstruation-inducing properties. It used to treat sore throat, fever, indigestion, and pains. Objectives: The objective of this study is to establish the chemical composition of the essential oil of flower, leaves of F. tingitana, and to throw light on antimicrobial, cytotoxic activities of Libyan plant. Materials and Methods: The chemical composition of the essential oil of flower (0.06% w/v) and leaves (0.1% w/v) of F. tingitana was comparatively analyzed by gas chromatography/mass spectrometry using nonpolar column DB-5. Results: A total of 28–32 components were identified, 15 being common in both samples. The main constituents of both flower- and leave-derived oil samples were α -thujene (13.5%–2.3%), elemol (8.9%–8.3%), eudesmol (0.6%–9.7%) and cadinol (2.2%–13.8%), respectively. The principle difference was a considerably more pronounced sesquiterpenes presence in the leaves-oil, amounting to 74.0%, than in the flower counterpart (39.9%). Caryophyllene (5.6%) and elemol (8.9%) were the major sesquiterpenes detected in flower-oil while leaves-oil showed less amounts of sesquiterpenoid hydrocarbons (27.7%) and represented by eudesmadiene (9.0%). On the contrary, while remaining the dominant group in both oil samples, monoterpenoids are relatively more abundant in flower-derived oil constituting 57.7% versus 24.5% detected in leaves. Conclusion: Leaves-oil sample being mostly efficient as antibacterial against Bacillus subtilis and Neisseria gonorrhoeae with potency 48.3, 41.9% compared to tetracycline standard antibacterial drug. The essential oil samples revealed marked in vitro cytotoxicity against breast (MCF7), cervical (HELA) and liver(HEPG2) carcinoma cell lines with IC50% (6.9, 4.8), (8.6, 10.9), and (4.4, 4.2) for the flower-, leaves-derived oil sample, respectively. Abbreviations used: F: Flower-derived oil of F. tingitana; L: Leaves-derived oil of F. tingitana ; IPP: Isopentenyl pyrophosphate or also isopentenyl diphosphate; DMAPP: Dimethylally pyrophosphate or also dimethylallyl diphosphate; GPP: Geranyl pyrophosphate; GGPP: Geranylgeranyl pyrophosphate; MEP: Methylerythritol phosphate pathway; FPP: Farnesyl pyrophosphate; GC/MS: Analysis gas chromatography/mass spectroscopy; SRB: Sulforhodamine B.
  2,209 114 6
Melastoma malabathricum ethyl acetate fraction induces secondary necrosis in human breast and lung cancer cell lines
Adi Idris, Ihsan N Zulkipli, Nurul Ramizah Zulhilmi, Huan F Lee, Rajan Rajabalaya, Lim Y Chee, Mohamed Majid, Sheba R David
July-September 2017, 13(51):688-692
DOI:10.4103/pm.pm_465_15  PMID:29142434
Background: Melastoma malabathricum (MM) is a traditional plant used in the Borneo region. The cytotoxic effects of methanol extracts from MM leaves have been reported in a number of human cancer cell lines. However, the mode of cell death by MM has not been investigated. Objective: We investigated the cytotoxic effects of MM in both human breast and lung cancer cell lines, MCF-7 and A549, respectively, and defined the mode of cell death. Materials and Methods: Cell viability was measured using the 3-(4-, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Annexin-V/propidium iodide (PI) and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining was done to determine the mode of cell death. Results: The MTT assay revealed that MM extract had an IC50 of >400 μ g/ml on both cell lines at 24 h posttreatment. Flow cytometric and fluorescence microscopy analysis of Annexin-V/PI stained MM-treated cells revealed that the majority of the cells underwent secondary necrosis/late apoptosis. TUNEL assay showed that little to no DNA nicks were present in MM-treated cells, suggesting that cells have undergone secondary necrosis, not late apoptosis, at that time point. Conclusion: MCF-7 and A549 cells undergoes secondary necrosis 24 h post-treatment with MM extract. MM leaf extract could be a potential source for a novel anti-tumor agent for cancer therapy. Abbreviations used: DMSO: Dimethyl sulfoxide; MM: Melastoma malabathricum; MTT: 3-(4-, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; PI: Propidium iodide; TUNEL: Terminal deoxynucleotidyl transferase dUTP nick-end labeling.
  2,179 107 2
Growth-arresting activity of acmella essential oil and its isolated component D-Limonene (1, 8 P-Mentha diene) against Trichophyton rubrum (Microbial type culture collection 296)
Diptikanta Padhan, Smaranika Pattnaik, Ajaya Kumar Behera
July-September 2017, 13(51):555-560
DOI:10.4103/pm.pm_65_17  PMID:29142414
Background: Spilanthes acmella is used as a remedy in toothache complaints by the tribal people of Western part of Odisha, India. Objective: The objective of this study was to study the growth-arresting activity of an indigenous Acmella essential oil (EO) (S. acmella Murr, Asteraceae) and its isolated component, d-limonene against Trichophyton rubrum (microbial type culture collection 296). Materials and Methods: The EO was extracted from flowers of indigenous S. acmella using Clevenger's apparatus and analyzed by gas chromatography–mass spectrometry (GC-MS). High pressure liquid chromatography (HPLC) was carried out to isolate the major constituent. The isolated fraction was subjected to fourier transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR). The antidermatophytic activity was screened for using “disc diffusion” and “slant dilution” method followed by optical, scanning electron microscopy (SEM), and transmission electron microscopy (TEM) studies. The molecular dockings were made between d-limonene with cell wall synthesis-related key enzymes (14 methyl deaminase and monooxygenase). Results: The GC-MS analysis EO had inferred the presence of 7 number of major (≥2%) components. The component with highest peak area (%) was found to be 41.02. The HPLC-isolated fraction was identified as d-limonene (1,8 p-Mentha-diene) by FTIR and NMR. Qualitative and quantitative assays had suggested the growth inhibitory activity of Acmella EO and its component. Shrinkage, evacuation, cell wall puncture, and leakage of cellular constituents by the activity of Acmella oil and d-limonene were evidenced from optical, SEM, and TEM studies. The computer simulation had predicted the binding strengths of d-limonene and fluconazole with dermatophyte cell wall enzymes. Conclusion: There could have been synergistic action of all or some of compounds present in indigenous Acmella EO. Abbreviations used: °C: Degree centigrade; w/v: Weight/volume; TS: Transverse section; min: minute; Hz: hertz: h: Hr.
  2,158 110 1
Investigation of antihyperglycaemic activity of banana (Musa sp. Var. Nanjangud rasa bale) flower in normal and diabetic rats
Ramith Ramu, Prithvi S Shirahatti, SP Dhanabal, Farhan Zameer, BL Dhananjaya, MN Nagendra Prasad
July-September 2017, 13(51):417-423
DOI:10.4103/0973-1296.216331  PMID:29142393
Background: The vital enzymes of starch digestion and absorption are intestinal α-glucosidases and their inhibition improves postprandial hyperglycaemia, constituting an effective mode of therapy in diabetes. Objectives: The present study was designed to assess the inhibitory potential of ethanol extract of banana flower (EF) on mammalian α-glucosidases and its pharmacological effects on postprandial hyperglycaemia in normal and alloxan-induced diabetic rats. Materials and Methods: EF was evaluated for its inhibitory potential and mode of inhibition on mammalian α-glucosidases. Further, the role of EF and its constituents Umbelliferone (C1) and Lupeol (C2) on glucose uptake using isolated rat hemi-diaphragm and insulinotropic activity using RINm5F (rat insulinoma) cell lines were determined. The phytocomponents in EF were also evaluated using GC-MS. Results: EF illustrated a dose-dependent inhibition for rat intestinal sucrase, maltase and p-nitrophenyl-α-D-glucopyranoside (pNPG) hydrolysis (IC50 values: 18.76±0.22, 25.54±0.10 and 76.42±1.12 μg/ml, respectively) and the mode of inhibition was non-competitive with low Ki values. Oral administration (100-200 mg/kg b.wt.) of EF significantly improved the maltose/glucose-induced postprandial hyperglycaemia in normal and alloxan-induced diabetic rats. EF, C1 and C2 exhibited stimulation of glucose uptake and a dose-dependent glucose-induced insulin secretion at both 4.5 and 16.7 mM glucose concentrations. Further, GC-MS analysis revealed significant levels of steroids (25.61%), diazoprogesterone (21.31%), sesquiterpene (11.78%) and other phytocomponents. Conclusion: EF inhibited α-glucosidases besides promoting glucose uptake and insulin secretion, resulting in antihyperglycaemic effect determining EF as a potent anti-diabetic agent. Abbreviations used: mg/dl: milligramsper deciliter, mM: millimolar, b.wt.: body weight.
  2,126 109 1
Comparative analysis of transcriptomes of macrophage revealing the mechanism of the immunoregulatory activities of a novel polysaccharide isolated from Boletus speciosus frost
Xiang Ding, Hongqing Zhu, Yiling Hou, Wanru Hou, Nan Zhang, Lei Fu
July-September 2017, 13(51):463-471
DOI:10.4103/pm.pm_151_16  PMID:28839373
Background: The mechanism of the immunoregulatory activities of polysaccharide is still not clear. Materials and Methods: Here, we performed the B-cell, T-cell, and macrophage cell proliferation, the cell cycle analysis of macrophage cells, sequenced the transcriptomes of control group macrophages, and Boletus speciosus Frost polysaccharide (BSF-1) group macrophages using Illumina sequencing technology to identify differentially expressed genes (DEGs) to determine the molecular mechanisms of immunomodulatory activity of BSF-1 in macrophages. Results: These results suggested that BSF-1 could promote the proliferation of B-cell, T-cell, and macrophages, promote the proliferation of macrophage cells by abolishing cell cycle arrests in the G0/G1 phases, and promote cell cycle progression in S-phase and G2/M phase, which might induce cell division. A total of 12,498,414 and 11,840,624 bp paired-end reads were obtained for the control group and BSF-1 group, respectively, and they corresponded to a total size of 12.5 G bp and 11.8 G bp, respectively, after the low-quality reads and adapter sequences were removed. Approximately 81.83% of the total number of genes (8,257) were expressed reads per kilobase per million mapped reads (RPKM ≥1) and more than 1366 genes were highly expressed (RPKM >60) in the BSF-1 group. A gene ontology-enrichment analysis generated 13,042 assignments to cellular components, 13,094 assignments to biological processes, and 13,135 assignments to molecular functions. A Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that the mitogen-activated protein kinase (MAPK) signaling pathways are significantly enriched for DEGs between the two cell groups. Conclusion: An analysis of transcriptome resources enabled us to examine gene expression profiles, verify differential gene expression, and select candidate signaling pathways as the mechanisms of the immunomodulatory activity of BSF-1. Based on the experimental data, we believe that the significant antitumor activities of BSF-1 in vivo mainly involve the MAPK signaling pathways. Abbreviations used: BSF-1: Boletus speciosus Frost polysaccharide.
  2,157 71 -
Extract of Bauhinia vahlii shows antihyperglycemic activity, reverses oxidative stress, and protects against liver damage in streptozotocin-induced diabetic rats
Ahmed H Elbanna, Mohammed M Nooh, Engy A Mahrous, Amal E Khaleel, Taha S Elalfy
July-September 2017, 13(51):607-612
DOI:10.4103/pm.pm_4_17  PMID:29142421
Background: Several studies have affirmed the effectiveness of some Bauhinia plants as antihyperglycemic agents. Objective: We investigated the possible effect of Bauhinia vahlii leaves extract in reducing hyperglycemia and reversing signs of organ damage associated with diabetes in streptozotocin (STZ) rat model. Materials and Methods: Both polar fraction of the B. vahlii leaves (defatted ethanolic extract [DEE]) and nonpolar fraction (n-hexane extract) were evaluated in vitro for α-glucosidase inhibition and 2,2-diphenyl-1-picrylhydrazyl radical scavenging potential. DEE was selected for further in vivo studies and was administered at two doses, i.e., 150 or 300 mg/kg to STZ-diabetic rats for 4 weeks. Results: Only DEE exhibited in vitro antioxidant and antihyperglycemic activities and its oral administration at both dose levels resulted in significant reduction in fasting blood glucose and glycated hemoglobin. Furthermore, signs of oxidative stress as indicated by hepatic reduced glutathione, nitric oxide, and malondialdehyde levels were completely reversed. In addition, histopathological examination and measurement of serum aspartate transaminase and alanine transaminase levels showed that DEE protected the liver from signs of liver pathogenesis when compared to diabetic untreated animals and those treated with metformin. Phytochemical analysis of DEE showed high flavonoids content with quercitrin as the major constituent along with other quercetin glycosides. Conclusion: This study strongly highlights the possible beneficial effect of B. vahlii leaves extract in relieving hyperglycemia and liver damage in STZ-diabetic rats and recommends further investigation of the value of quercetin derivatives in controlling diabetes and ameliorating liver damage associated with it. Abbreviations used: ALT: Alanine transaminase, AST: Aspartate transaminase, DEE: Defatted ethanol extract, DPPH: 2,2-diphenyl-1-picrylhydrazyl, FBG: Fasting blood glucose, GAE: Gallic acid equivalent, GSH: Reduced glutathione, Hb1Ac: Glycated hemoglobin, HE: Hexane extract MDA: Malondialdehyde, QE: Quercetin equivalent, STZ: Streptozotocin, TAC: Total antioxidant capacity.
  2,128 97 2
Marantodes pumilum (Blume) kuntze inhibited secretion of lipopolysaccharide- and monosodium urate crystal-stimulated cytokines and plasma prostaglandin E2
Eldiza Puji Rahmi, Jamia Azdina Jamal, Endang Kumolosasi, Juriyati Jalil, Nor-Ashila Aladdin
July-September 2017, 13(51):578-586
DOI:10.4103/pm.pm_35_17  PMID:29142418
Background: Marantodes pumilum is traditionally used for dysentery, gonorrhea, and sickness in the bones. Previous studies revealed its antibacterial and xanthine oxidase inhibitory activities. Objective: To evaluate the inhibitory effects of three M. pumilum varieties on the secretion of lipopolysaccharide (LPS)- and monosodium urate crystal (MSU)-induced cytokines and plasma prostaglandin E2 (PGE2) in vitro. Materials and Methods: The leaves and roots of M. pumilum var. alata (MPA), M. pumilum var. pumila (MPP), and M. pumilum var. lanceolata (MPL) were successively extracted with dichloromethane (DCM), methanol, and water. Human peripheral blood mononuclear cells and ELISA technique were used for the cytokine assay, whereas human plasma and radioimmunoassay technique were used in the PGE2assay. Flavonoids content was determined using a reversed-phase high-performance liquid chromatography. Results: DCM extract of MPL roots showed the highest inhibition of LPS-stimulated cytokine secretion with IC50values of 29.87, 7.62, 5.84, 25.33, and 5.40 μ g/mL for interleukin (IL)-1α , IL-1β , IL-6, IL-8, and tumor necrosis factor (TNF)-α , respectively; while that of plasma PGE2 secretion was given by DCM extract of MPP roots (IC50 31.10 μ g/mL). Similarly, the DCM extract of MPL roots demonstrated the highest inhibition against MSU-stimulated IL-1α , IL-1β , IL-6, IL-8, TNF-α , and PGE2 secretion with IC50values of 11.2, 8.92, 12.29, 49.51, 9.60, and 31.58 μ g/mL, respectively. Apigenin in DCM extracts of MPL (0.051 mg/g) and MPP (0.064 mg/g) roots could be responsible for the strong inhibitory activity against IL-1β , IL-6, TNF-α , and PGE2. Conclusion: The results suggested that DCM extracts of MPL and MPP roots are potential anti-inflammatory agents by inhibiting the secretion of LPS- and MSU-stimulated pro-inflammatory cytokines and PGE2.
  2,039 152 1
Quantitative analysis of benzyl isothiocyanate in Salvadora persica extract and dental care herbal formulations using reversed phase C18 high-performance liquid chromatography method
Maged Saad Abdel-Kader, YT Kamal, Prawez Alam, Mohammed Hamed Alqarni, Ahmed Ibrahim Foudah
July-September 2017, 13(51):412-416
DOI:10.4103/pm.pm_117_17  PMID:29142392
Context: Benzyl isothiocyanate is the active antimicrobial agent in Salvadora persica (siwak) widely used in Islamic countries for oral hygiene. Aims: Quantification of benzyl isothiocyanate in the ethanol extract of S. persica and some dental care herbal formulations labeled to contain siwak. Settings and Design: Simple and sensitive high-performance liquid chromatography method was designed. Subjects and Methods: Separation was achieved on reverse phase C18 (250 mm × 4.6 mm, 5 μ ) column with a mobile phase comprising acetonitrile and water (1:1). The detection was carried out at 190 nm using ultra violet-visible detector. The flow rate was kept at 1 mL/min. Results: A sharp and well-defined peak was obtained at the retention time of 9.322 ± 0.3 min. Linear regression analysis data for the calibration plot showed a good linear relationship between response and concentration in the range of 0.5–500 μg/mL with a regression coefficient (r2) of 0.9977. The method was validated for accuracy, precision, robustness, and sensitivity. All the parameters examined met the current recommendations for the International Conference on Harmonization guidelines for method validation. Conclusions: The method was applied for the quantification of benzyl isothiocyanate in siwak extract, dental care powder, mouth wash, and toothpaste claimed to contain siwak. The developed method was found specific, simple, selective, and reliable for routine use in quality control analysis of different commercially available herbal care products. Abbreviations used: RP18: Reversed phase C18, HPLC: High performance liquid chromatography, UV: Ultra violet, r2: regression coefficient, ICH: international conference on harmonization, TLC: Thin layer chromatography, CHCl3: Chloroform, v/v: volume/volume, RSD: Relative standard deviation, LOD: Limit of detection, LOQ: Limit of quantification.
  2,016 106 2
In vitro protoscolicidal effects of Cinnamomum zeylanicum essential oil and its toxicity in mice
Hossein Mahmoudvand, Hormoz Mahmoudvand, Razieh Tavakoli Oliaee, Amir Tavakoli Kareshk, Seyed Reza Mirbadie, Mohammad Reza Aflatoonian
July-September 2017, 13(51):652-657
DOI:10.4103/pm.pm_280_16  PMID:29142428
Background: This study investigates the scolicidal effects of Cinnamomum zeylanicum essential oil against the protoscoleces of hydatid cysts and its toxicity in the mice model. Materials and Methods: Gas chromatography/mass spectroscopy analyses were used to identify the constituents of essential oil. Protoscoleces were treated with different concentrations of the essential oil (6.25–100 μL/mL) in each test tube for 5–30 min. The viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Forty-eight male NMRI mice were also used to determine the toxicity of C. zeylanicum essential oil (0.5–4 mL/kg). Results: The main components were found to be cinnamaldehyde (91.8%), ρ metoxicinamate (1.57%), and α pinene (1.25%). Findings indicate that C. zeylanicum essential oil with the concentrations of 100 and 50 μL/mL killed 100% of protoscoleces after 5 min of exposure. Also, the lower concentrations of C. zeylanicum essential oil motivated a late protoscolicidal effect. The LD50value of intraperitoneal injection of C. zeylanicum essential oil was 2.07 mL/kg body weight after 48 h, and the maximum nonfatal dose was 1.52 mL/kg body weight. The results also showed that there was no significant toxicity following oral administration of C. zeylanicum essential oil for 2 weeks. Conclusion: The results exhibited the favorable scolicidal activity of C. zeylanicum, which could be applied as a natural scolicidal agent in hydatid cyst surgery.
  2,000 119 2
Characterization of leaf extracts of Schinus terebinthifolius raddi by GC-MS and chemometric analysis
Fabíola B Carneiro, Pablo Q Lopes, Ricardo C Ramalho, Marcus T Scotti, Sócrates G Santos, Luiz A. L. Soares
July-September 2017, 13(51):672-675
DOI:10.4103/pm.pm_555_16  PMID:29142431
Background: Schinus terebinthifolius Raddi belongs to Anacardiacea family and is widely known as “aroeira.” This species originates from South America, and its extracts are used in folk medicine due to its therapeutic properties, which include antimicrobial, anti-inflammatory, and antipyretic effects. The complexity and variability of the chemical constitution of the herbal raw material establishes the quality of the respective herbal medicine products. Objective: Thus, the purpose of this study was to investigate the variability of the volatile compounds from leaves of S. terebinthifolius. Materials and Methods: The samples were collected from different states of the Northeast region of Brazil and analyzed with a gas chromatograph coupled to a mass spectrometer (GC-MS). The collected data were analyzed using multivariate data analysis. Results: The samples' chromatograms, obtained by GC-MS, showed similar chemical profiles in a number of peaks, but some differences were observed in the intensity of these analytical markers. The chromatographic fingerprints obtained by GC-MS were suitable for discrimination of the samples; these results along with a statistical treatment (principal component analysis [PCA]) were used as a tool for comparative analysis between the different samples of S. terebinthifolius. Conclusion: The experimental data show that the PCA used in this study clustered the samples into groups with similar chemical profiles, which builds an appropriate approach to evaluate the similarity in the phytochemical pattern found in the different leaf samples. Abbreviations used: AL: Alagoas, BA: Bahia, CE: Ceará, CPETEC: Center for Weather Forecasting and Climate Studies, GC-MS: Gas chromatograph coupled to a mass spectrometer, MA: Maranhão, MVA: Multivariate data analysis, PB: Paraíba, PC1: Direction that describes the maximum variance of the original data, PC2: Maximum direction variance of the data in the subspace orthogonal to PC1, PCA: Principal component analysis, PE: Pernambuco, PI: Piauí, RN: Rio Grande do Norte, SE: Sergipe.
  1,926 130 -
Chemotaxonomic diversity of three Ficus species: Their discrimination using chemometric analysis and their role in combating oxidative stress
Nawal Al-Musayeib, Sherif S Ebada, Haidy A Gad, Fadia S Youssef, Mohamed Lotfy Ashour
July-September 2017, 13(51):613-622
DOI:10.4103/pm.pm_579_16  PMID:29142422
Background: Genus Ficus (Moraceae) constitutes more than 850 species and about 2000 varieties and it acts as a golden mine that could afford effective and safe remedies combating many health disorders. Objectives: Discrimination of Ficus cordata, Ficus ingens, and Ficus palmata using chemometric analysis and assessment of their role in combating oxidative stress. Materials and Methods: Phytochemical profiling of the methanol extracts of the three Ficus species and their successive fractions was performed using high-performance liquid chromatography/electrospray ionization mass spectrometry. Their discrimination was carried out using the obtained spectral data applying chemometric unsupervised pattern-recognition techniques, namely, principal component analysis and hierarchical cluster analysis. In vitro hepatoprotective and antioxidant evaluation of the samples was performed using human hepatocellular carcinoma cells challenged by carbon tetrachloride (CCl4). Results: Altogether, 22 compounds belonging to polyphenolics, flavonoids, and furanocoumarins were identified in the three Ficus species. Aviprin is the most abundant compound in F. cordata while chlorogenic acid and psoralen were present in high percentages in F. ingens and F. palmata, respectively. Chemometric analyses showed that F. palmata and F. cordata are more closely related chemically to each other rather than F. ingens. The ethyl acetate fractions of all the examined species showed a marked hepatoprotective efficacy accounting for 54.78%, 55.46%, and 56.42% reduction in serum level of alanine transaminase and 56.82%, 54.16%, and 57.06% suppression in serum level of aspartate transaminase, respectively, at 100 μ g/mL comparable to CCl4-treated cells. Conclusion: Ficus species exhibited a notable antioxidant and hepatoprotective activity owing to their richness in polyphenolics and furanocoumarins. Abbreviations used: ALT: Alanine transaminase, AST: Aspartate transaminase, CCl4:Carbon tetrachloride, DMEM: Dulbecco's Modified Eagle's medium, DMSO: Dimethyl sulfoxide, EDTA: Ethylenediaminetetraacetic acid, FBS: Fetal bovine serum, FCA: Ficus cordata remaining aqueous fraction, FCB: Ficus cordata n-butanol fraction, FCE: Ficus cordata ethyl acetate fraction, FCP: Ficus cordata petroleum ether fraction, FCT: Ficus cordata total methanol extract, FIA: Ficus ingens remaining aqueous fraction, FIB: Ficus ingens n-butanol fraction, FIE: Ficus ingens ethyl acetate fraction, FIP: Ficus ingens petroleum ether fraction, FIT: Ficus ingens total methanol extract, FPA: Ficus palmata remaining aqueous fraction, FPB: Ficus palmata n-butanol fraction, FPE: Ficus palmata ethyl acetate fraction, FPP: Ficus palmata petroleum ether fraction, FPT: Ficus palmata total methanol extract, GSH: Reduced glutathione,HepG2 cells: Human hepatocellular carcinoma, HPLC-ESI-MS: High-performance liquid chromatography/electrospray ionization mass spectrometry, and SOD: Superoxide dismutase.
  1,902 102 1
Flavonoidal constituents, antioxidant, antimicrobial, and cytotoxic activities of Dipterygium glaucum grown in Kingdom of Saudi Arabia
Usama Shaheen, Nagwa Abdelkader Shoeib, Abeer Temraz, Mohamed I. S. Abdelhady
July-September 2017, 13(51):484-488
DOI:10.4103/pm.pm_44_16  PMID:29142403
Background: Dipterygium glaucum Decne. herb is one of the common traditional plants with multiple medicinal uses. Objective: To isolate the major constituents and to investigate the antioxidant, antimicrobial, and cytotoxic activities of this herb. Materials and Methods: Methanolic extract of D. glaucum herb was fractionated using n-hexane, dichloromethane, and n-butanol. Butanol fraction was chromatographed using column chromatography and preparative thin layer chromatography to isolate the major constituents. Isolated compounds were elucidated by means of spectroscopic methods, including 1D, 2D NMR (1H, 13C, DEPT, COSY, HSQC, HMBC, NEOSY) and MS analysis. Total phenolic content using Folin–Ciocalteu reagent and antioxidant activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay of the total methanolic extract were evaluated. Cytotoxic potential of both methanolic extract and butanol fraction was tested using a crystal violet viability assay. Antimicrobial activities of both extracts were investigated using diffusion agar technique. Results: Apigenin 6, 8-di-C-glucopyranoside (vicenin-2), quercetin-3'-O-methyl-3-O-glucopyranoside, quercetin-3'-O-methyl-3-O-galactopyranoside, quercetin-3-O-β-D-glucopyranoside, and quercetin-3-O-β-D-galactopyranoside were isolated and elucidated. Total phenolic content was (83.89 mg gallic acid equivalent/g extract). The EC50value of scavenging DPPH radical was 152.0 ± 2 μ g/mL. Butanol fraction showed the highest cytotoxic activity against cervical and breast carcinoma cells (IC50 3.6 and 6.1 μ g/mL, respectively). Both methanolic extract and butanol fraction showed wide spectrum antimicrobial activity against Gram-positive and Gram-negative bacteria and some fungi. The highest activity was from methanolic extract against Enterococcus faecalis (83.25%) and against Candida tropicalis (77.03%) as compared to reference antibiotics. Conclusion: Data obtained from this study demonstrate that D. glaucum possesses significant antioxidant, cytotoxic, and antimicrobial activities which could be ascribed to its flavonoidal content. Abbreviations used: KSA: Kingdom of Saudi Arabia; TLC: Thin Layer Chromatography; DPPH: 2,2'-diphenyl-1-picrylhydrazyl; EC50: Half maximal effective concentration; IC50: Half maximal inhibitory concentration; DMSO: dimethyl sulfoxide; NMR: Nuclear Magnetic Resonance; ESIMS: Electrospray ionization mass spectrometry; MeOH: Methyl alcohol.
  1,890 106 2
Prosopis juliflora pods alkaloid-rich fraction: In vitro anthelmintic activity on goat gastrointestinal parasites and its cytotoxicity on vero cells
Helimar Gonçalves Lima, Danilo Cavalcante Gomes, Nathália Silva Santos, Êuder Reis Dias, Mariana Borges Botura, Maria José Moreira Batatinha, Alexsandro Branco
July-September 2017, 13(51):684-687
DOI:10.4103/pm.pm_3_17  PMID:29142433
Background: This study was designed to assess the in vitro anthelmintic activity of the fraction containing alkaloid from Prosopis juliflora pods on goat gastrointestinal nematodes using the egg hatch assay (EHA), larval migration inhibition assay (LMIA), and larval motility assay (LMA). Materials and Methods: The alkaloid-rich fraction (AF) – content juliprosopine as major alkaloid – was obtained from ethyl acetate extract after fractionation in Sephadex LH-20 chromatography column and its characterization were made by nuclear magnetic resonance analysis together with literature data comparison. The concentrations tested were 4.0, 2.67, 1.78, 1.19, and 0.79 mg/mL (EHA) and 4 mg/mL (LMIA and LMA). The in vitro cytotoxicity on Vero cell cultures was determined with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue tests. Results: High ovicidal activity was observed with IC50and IC90 values at 1.1 and 1.43 mg/mL for AF. On the other hand, this fraction showed low larvicidal activity and high toxic effect. Conclusion: Thus, P. juliflora pod alkaloid rich-fraction has ovicidal activity in vitro against goat gastrointestinal nematodes and cytotoxic in Vero cell cultures. Abbreviations used: AF: Alkaloid-rich fraction; DMSO: Dimethyl sulfoxide; EE: Ethyl acetate extract; EHA: Egg hatch assay; IC50: Inhibitory concentration 50%; IC90: Inhibitory concentration 90%; L3: Infective larvae; LMA: Larval motility assay; LMIA: Larval migration inhibition assay; MTT: Bromide 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; NMR: Nuclear magnetic resonance; PBS: Phosphate buffered saline; RPMI: Roswell Park Memorial Institute médium; TLC: Thin Layer Chromatography.
  1,897 94 2
Screening of norharmane from seven cyanobacteria by high-performance liquid chromatography
Tunay Karan, Ramazan Erenler
July-September 2017, 13(51):723-725
DOI:10.4103/pm.pm_214_17  PMID:29142439
Background: Cyanobacteria, including pharmaceutically and medicinally valuable compounds attract the great attention lately. Norharmane (9H-pyrido (3,4-b) indole found in some cyanobacteria revealed a great number of biological effects. Objective: Seven cyanobacteria were isolated and identified from Yesilirmak River and Gaziosmanpasa University Campus to determine the norharmane content. Materials and Methods: Cyanobacteria collected from Tokat, Turkey were isolated and identified by morphologically. Norharmane (9H-pyrido [3,4-b] indole) quantities were presented for seven cyanobacteria, Chroococcus minutus (Kütz.) Nägeli, Geitlerinema carotinosum (Geitler) Anagnostidis, Nostoc linckia Bornet ex Bornet and Flahault, Anabaena oryzae F. E. Fritsch, Oscillatoria limnetica Lemmermann, Phormidium sp. Kützing ex Gomont, and Cylindrospermum sp. Kutzing ex E. Bornet and C. Flahault by high-performance liquid chromatography. Results: The norharmane amount indicated for cyanobacterial culture media altered in a species-dependent kind in the range of 0.81–10.87 μ g/g. C. minutus produced the most norharmane among the investigated cyanobacteria as 10.87 μ g/g. Conclusion: Cyanobacteria could be an important source of norharmane as well as pharmaceutically valuable compounds. Abbreviations used: HPLC: High performance liquid chromatograph.
  1,817 108 2
Constituents and antioxidant activity of bleeding sap from various Xinjiang grapes
Lv Le, Anwar Umar, Arkin Iburaim, Nicholas Moore
July-September 2017, 13(51):726-730
DOI:10.4103/pm.pm_358_16  PMID:29142440
Objective: Wine grape sap or bleeding sap of grapes (GBS) is commonly used in Xinjiang (China) for therapeutic aims. Do variations in composition related to region and variety affect its properties? Methods: GBS samples originating in various parts of Xinjiang (Turpan, Hotan, Kashgar, and Atush) were tested for phenols and polyphenols, polysaccharides, saponin, proteins, individual amino acids, and minerals. Their antioxidant activity was measured using ascorbic acid as reference. Results: Polyphenol content varied from 2.6 to 6.6 mg/L, polysaccharides 18.3–816 mg/L, saponin 6.25–106 mg/L, and protein 3.0–22.4 mg/L. Mineral elements and amino acids ranged from 6.20 to 201.2 mg/L and 0.06–118.7 mg/L, respectively. ·OH scavenging ability varied from 70% to over 90%, higher than Vitamin C. Grapes from Turpan had lower antioxidant activity than other grapes even though the polyphenol content was generally higher. Conclusion: Bleeding sap of Xinjiang grape is rich in amino acids, polysaccharides, polyphenols, and protein. The contents are different according to the origin, related possibly to species, climate, and environment. Antioxidant effects were not correlated with polyphenol content. Abbreviations used: GBS: Bleeding sap of grapes; PITC: phenyl isothiocyanate.
  1,820 90 1
High-performance thin-layer chromatographic-densitometric quantification and recovery of bioactive compounds for identification of elite chemotypes of Gloriosa superba L. collected from Sikkim Himalayas (India)
Ankita Misra, Pushpendra Kumar Shukla, Bhanu Kumar, Jai Chand, Poonam Kushwaha, Md Khalid, Ajay Kumar Singh Rawat, Sharad Srivastava
July-September 2017, 13(51):700-705
DOI:10.4103/pm.pm_576_16  PMID:29142436
Background: Gloriosa superba L. (Colchicaceae) is used as adjuvant therapy in gout for its potential antimitotic activity due to high colchicine(s) alkaloids. Objective: This study aimed to develop an easy, cheap, precise, and accurate high-performance thin-layer chromatographic (HPTLC) validated method for simultaneous quantification of bioactive alkaloids (colchicine and gloriosine) in G. superba L. and to identify its elite chemotype(s) from Sikkim Himalayas (India). Methods: The HPTLC chromatographic method was developed using mobile phase of chloroform: acetone: diethyl amine (5:4:1) at λ maxof 350 nm. Results: Five germplasms were collected from targeted region, and on morpho-anatomical inspection, no significant variation was observed among them. Quantification data reveal that content of colchicine (Rf: 0.72) and gloriosine (Rf: 0.61) varies from 0.035%–0.150% to 0.006%–0.032% (dry wt. basis). Linearity of method was obtained in the concentration range of 100–400 ng/spot of marker(s), exhibiting regression coefficient of 0.9987 (colchicine) and 0.9983 (gloriosine) with optimum recovery of 97.79 ± 3.86 and 100.023% ± 0.01%, respectively. Limit of detection and limit of quantification were analyzed, respectively, as 6.245, 18.926 and 8.024, 24.316 (ng). Two germplasms, namely NBG-27 and NBG-26, were found to be elite chemotype of both the markers. Conclusion: The developed method is validated in terms of accuracy, recovery, and precision studies as per the ICH guidelines (2005) and can be adopted for the simultaneous quantification of colchicine and gloriosine in phytopharmaceuticals. In addition, this study is relevant to explore the chemotypic variability in metabolite content for commercial and medicinal purposes.
  1,789 89 3
The effect of thymoquinone on nuclear factor kappa B levels and oxidative DNA damage on experimental diabetic rats
Ayşe Usta, Semiha Dede
July-September 2017, 13(51):458-461
DOI:10.4103/pm.pm_134_17  PMID:29142399
Background: Thymoquinone (TQ), the basic bioactive phytochemical constituent of seed oil of Nigella sativa, is one of these herbal drugs known for antidiabetic effects. This study was carried out to assess the effects of the possible role of TQ on nuclear factor kappa B (NF-κB) and oxidative DNA damage levels in experimental diabetic rats. Materials and Methods: Twenty-eight male Wistar Albino rats (200–250 g) were used as experimental subjects. The rats were divided into four groups, including the control, control supplemented with TQ (CT), diabetic (D), and diabetic supplemented with TQ (DT), each containing seven rats. The D and the DT groups were treated with 45 mg/kg streptozotocin (STZ) (intraperitoneal). TQ was administered 30 mg/kg/day for 21 days by oral gavage in the DT and the T groups. Results: It was determined that glucose, glycosylated hemoglobin (HbA1c) levels and alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyl transpeptidase activities were decreased significantly and approached the control group in the DT group after TQ supplement (P < 0.05). Urea levels were the lowest in CT (P < 0.05). Oxidative DNA damage (8 hydroxy-2-deoxyguanosine) was increased in both of the diabetic groups (D and DT). The NF-κB levels were the highest in Group D (P < 0.05). Conclusion: It was observed that increased glucose and HbA1c levels and the indicators of liver and kidney damages were decreased significantly after TQ supplementation. Oxidative DNA damage and NF-κB levels were increased in the diabetic group, and TQ administration caused a statistically insignificant reduction. Abbreviations used: 8-OHdG: 8 hydroxi-2-deoxiguanosin; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; GGT: Gamma-glutamyl transpeptidase; HbA1c: Glycosylated hemoglobin; NF-κB: Nuclear factor kappa protein; STZ: Streptozotocin; TQ: Thymoquinone.
  1,713 100 3
The effect of polyherbal medicines used for the treatment of tuberculosis on other opportunistic organisms of humans infected with tuberculosis
Elizabeth Bosede Famewo, Anna Maria Clarke, Anthony Jide Afolayan
July-September 2017, 13(51):539-543
DOI:10.4103/pm.pm_468_16  PMID:29142411
Background: In many immunocompromised patients, opportunistic bacterial and fungal infections are common. Polyherbal medicines examined in this study are used by the indigenous people of South Africa for the treatment of tuberculosis (TB) and other opportunistic infections associated with TB. Objective: To evaluate the antibacterial and antifungal activity of nine polyherbal remedies against four Gram-positive and Gram-negative bacteria respectively and three fungi. Materials and Methods: Agar dilution method was used to determine the minimum inhibitory concentration (MIC) of the remedies against the organisms. Results: The inhibitory activity of the polyherbal medicines based on the overall MIC revealed that HBfs and FB remedies were the most active remedies against the bacterial isolates at the concentration of 2.5 mg/mL, followed by HBts remedy at 5.0 mg/mL. However, the MIC valves of KWTa, KWTb, KWTc, HBss, EL and AL remedies were higher than 5.0 mg/mL which was the highest concentration used. Only KWTa remedy showed activity against Aspergillus niger and Aspergillus fumigatus with the MIC value of 2.5 mg/mL. While KWTc and HBts had the highest activity at 1.25 mg/mL against Candida albicans, the remaining remedies were active at 2.5 mg/mL. Conclusion: This study revealed that some of these polyherbal formulations have activities against some of the opportunistic bacterial and fungal isolates associated with TB patients. The capability of these remedies to inhibit the organisms is an indication that they are a potential broad-spectrum antimicrobial agent. However, the remedies that are inactive might contain stimulant effects on the immune system. Abbreviations used: TB: Tuberculosis; MIC: Minimum Inhibitory Concentration; CFU/ML: Colony Forming
  1,704 85 -
Inhibition of phosphorylated c-Jun NH(2)-terminal kinase by 2',4'-dihydroxy-6-methoxy-3,5-dimethylchalcone isolated from Eugenia aquea Burm f. leaves in jurkat T-cells
Melisa I Barliana, Ajeng Diantini, Anas Subarnas, Rizky Abdulah, Takashi Izumi
July-September 2017, 13(51):573-577
DOI:10.4103/pm.pm_16_17  PMID:29142417
Background: Indonesian medicinal plants have been used for their anticancer activity for decades. However, the therapeutic effects of medicinal plants have not been fully examined scientifically. As cancer is a major health problem worldwide, searching for a new anticancer compound has attracted considerable attention. Our previous study found that 2',4'-dihydroxy-6-methoxy-3,5-dimethylchalcone, an active compound isolated from leaves of Indonesian medicinal plants Eugenia aquea Burm f. (Myrtaceae), had anticancer activity in MCF-7 human breast cancer cells through induction of apoptosis. Objective: To investigate the molecular mechanism of 2',4'-dihydroxy-6-methoxy-3,5-dimethylchalcone antiproliferative activity. Materials and Methods: Leaves of E. aquea were extracted by ethanol, fractionated by ethyl acetate, n-hexane, or water, and isolated for its active compound. Jurkat T-cells were treated with 2',4'-dihydroxy-6-methoxy-3,5-dimethylchalcone for 12 and 24 h, and a cell viability assay and real-time-reverse transcriptase polymerase chain reaction for interleukin-2 (IL-2) mRNA measurement were performed. The effects of active compound to mitogen-activated protein kinases were also examined to investigate the mechanism of its antiproliferative activity. Results: 2',4'-dihydroxy-6-methoxy-3,5-dimethylchalcone inhibited Jurkat T-cell proliferation with a half maximal inhibitory concentration of 59.5 μ M. Although IL-2 mRNA expression was slightly increased after treatment, it inhibited c-Jun N-terminal kinase expression but not p38 and extracellular signal-regulated kinase expression. Conclusions: Our study indicated that the molecular mechanism mediating the antiproliferative activity of 2',4'-dihydroxy-6-methoxy-3,5-dimethylchalcone may be attributed to the stimulation of an immunological microenvironment in the cells. Abbreviations used: E. aquea: Eugenia aquea, IL-2: Interleukin-2, MAPK: Mitogen-activated protein kinase, ERKs: Extracellular signal-regulated kinases, JNKs: c-Jun N-terminal kinases, p38: p38 MAPK, PI3K: Phosphatidylinositol-3 kinase, IC50: Half maximal inhibitory concentration.
  1,665 82 -
Chromosomal fragmentation: A possible marker for the selection of high gymnemic acid yielding accessions of Gymnema sylvestre R. Br
Ashutosh Kumar Verma, Sunita Singh Dhawan
July-September 2017, 13(51):481-483
DOI:10.4103/pm.pm_420_16  PMID:29142402
Background: Gymnema sylvestre R. Br. a member of family Asclepiadaceae as mentioned in Indian Pharmacopoeia popular among the researchers because of stimulatory effect of its phytoconstituent on pancreatic cells and potential to treat Type I and II type of diabetes. Objectives: Development of cost-effective marker system for the selection of high gymnemic acid yielding accessions of G. sylvestre. Materials and Methods: Presoaked seeds of Brassica campestris treated with different dilutions of gymnemagenin and 10% leaf extract of twenty different accessions of G. sylvestre. Root tips of germinated seeds were fixed, and chromosomal studies were made by root tip bioassay method. Results: Exposure of seeds to treatment solutions promotes various types of chromosomal anomalies in root meristem, and surprisingly, direct correlation between the percentage of chromosomal fragmentation and the percentage of gymnemic acid shared by treatment solution were observed. Conclusion: Later finding may be explored for the development of a novel methodology or marker system for the selection of high active principle yielding accessions of G. sylvestre. Abbreviations used: MI: Mitotic index; CP: Condensed prophase; CM: Clumped metaphase; MC: Metaphase cleft; FR: Fragmentation; AP: Anaphase with persistent nucleolous; LA: Laggard, BR: Bridge; BI: Bi-nucleated cell; DA: Disturbed anaphasic polarity.
  1,592 68 -
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