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January-March 2019
Volume 15 | Issue 60
Page Nos. 1-176

Online since Wednesday, January 23, 2019

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ORIGINAL ARTICLES  

Picrolemma sprucei quassinosides inhibits breast and prostate cell growth and impairs behavioral phenotype in mice Highly accessed article p. 1
Sinária Rejany Nogaia de Sousa, Ingrit Elida Collantes Díaz, Riad Naim Younes, Sergio Alexandre Frana, Maria Martha Bernardi, Ivana Barbosa Suffredini
DOI:10.4103/pm.pm_228_18  
Background: It is estimated that in Brazil 59,700 new cases of breast and 68,220 new cases of prostate cancer will occur, in 2018, in Brazil. For that reason, there is a need for introducing new antitumor drugs in therapy. Objective: The objective of the study is to bioguide fractionate the organic/aqueous extracts from the aerial organs of Picrolemma sprucei (EB1151 and EB1152) aiming the identification of antitumor compounds and to evaluate the influence of EB1151 on mice behavioral phenotype. Materials and Methods: The breast (MCF-7) and prostate (PC-3) cancer cell lines cytotoxicity assay were assessed using the sulforhodamine B method. Fractionation was carried out by thin layer and column chromatographic techniques. The influence of the intraperitoneal administration of nonlethal dose (NLD, 39.1 mg/kg) of EB1151 to Balb-c male mice was assessed in open cage and open-field (OF) apparatuses. Results: FCHCl3partition phases from EB1151 and EB1152 showed cytotoxic activity against both cancer cells, as did fractions UNIP343, which contains sergeolide and isobruceine B, and UNIP344, which contains sergeolide and 15-deacetylsergeolide. Higher doses of EB1151 showed significant alterations in response to touch, hindquarter fall, irritability, auricular and corneal reflexes, ataxia, micturition, ptosis, piloerection, hypothermia, and cyanosis, and NLD showed alterations in general activity and response to touch, but recovery was obtained up to the end of the experiment. Conclusions: Quassinosides from the aerial organs of P. sprucei showed significant cytotoxicity against MCF-7 and PC-3 cell lines. Extract EB1151 has impaired general activity and activity in the OF apparatus, but recovery was observed. Abbreviations used: IBAMA/CGen/MMA: Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis/Conselho de Gestão do Patrimônio Genético/Ministério do Meio Ambiente; UNIP: Universidade Paulista; g: Gram; h: Hour; mL: Milliliters; FCHCl3: Fraction chloroform; FBuOH: Fraction butanol; FH2O: Fraction water; FHEX: Fraction hexane; FDCM: Fraction dichloromethane; FMeOH: Fraction methanol; FACN15: Fraction acetonitrile 15%; FACN50: Fraction acetonitrile 50%; FAcEt: Fraction ethyl acetate; CDCl3: Deuterated chloroform; NH4OH: Amonium hydroxide; DOXO: Doxorubicin; M: Molar; TCA: Trichloroacetic acid; μg: Microgram; IC50: 50% inhibitory concentration; CEUA/ICS/UNIP: Comissão de Ética no Uso de Animais/Instituto de Ciências da Saúde/Universidade Paulista; kg: cm3: Cubic centimeters; Kilogram; BP: Behavioral phenotype; OF: Open-field apparatus; NLD: Nonlethal dose; mL: Microliters; ANOVA: Analysis of variance; FAkl: Fraction alkaloids; mM: Millimolar; DNA: Desoxyribonucleic acid.
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Cytotoxicity and mitochondrial-mediated apoptosis induced by Fenugreek seed oil in human hepatocellular carcinoma cells via reactive oxygen species generation Highly accessed article p. 12
Ebtesam S Al-Sheddi, Nida N Farshori, Mai M Al-Oqail, Shaza M Al-Massarani, Maqsood A Siddiqui, Javed Ahmad, Abdulaziz A Al-Khedhairy
DOI:10.4103/pm.pm_163_18  
Background: Cancer has been recognized as a major cause of death globally. Traditional medicines have been used to treat cancer for several periods, and herbal medicines are currently being used for the treatment of cancer worldwide. Fenugreek is an annual plant (family: Fabaceae) and is documented as a medicinal herb since olden days. Objective: The present study was aimed to investigate the anticancer activity of Fenugreek seed oil (FSO) against human hepatocellular carcinoma (HepG2) cell line. Materials and Methods: The HepG2 cells were exposed to 10–1000 μg/ml of FSO for 24 h to assess the cytotoxicity using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide test, neutral red uptake assay, and morphological changes. The cytotoxic concentrations of FSO were used to study the induction in reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) levels. Further, the expression of pro-and antiapoptotic genes (p53, caspase-3, caspase-9, Bax and Bcl-2) were studied. Results: A concentration-dependent decrease in cell viability of HepG2 cells was observed after FSO exposure. Our results showed that FSO decreased cell viability of HepG2 cells in a concentration-dependent manner. FSO was also found to increase the ROS generation and decrease the MMP level in HepG2 cells. Our gene expression study indicates upregulation in apoptotic marker genes and downregulation in antiapoptotic gene. Conclusion: The results showed that FSO is capable to induce mitochondrial-mediated apoptosis through ROS generation in HepG2 cells and could be a useful resource in developing effective remedies for the treatment of hepatocellular carcinoma. Abbreviations used: FSO: Fenugreek seed oil; HepG2: Human hepatocellular carcinoma; ROS: Reactive oxygen species; NRU: Neutral red uptake; DMSO: Dimethyl sulfoxide; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; MMP: Mitochondrial membrane potential; DNA: Deoxyribonucleic acid; RNA: Ribonucleic acid; DCFH-DA: 2,7-dichlorofluorescein diacetate; FBS: Fetal bovine serum; PBS: Phosphate-buffered saline; PCRq: quantitative real-time PCR; NaHCO3: Sodium bicarbonate; CO2: Carbon dioxide; DMEM: Dulbecco's modified eagle medium.
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Differential gene expressions and phytohormone changes altered Lonicera japonica quality after plant introduction p. 18
Xiaoying Chen, Shuangshuang Qin, Cui Li, Qinghua Wu, Chao Jiang, Jian Yang, Xiaoyun Guo, Chunli Ou
DOI:10.4103/pm.pm_317_18  
Background: Lonicera japonica Thunb. (LJ) is a famous traditional Chinese medicinal plant. For great viability and important medical value, LJ was introduced from the geo-authentic areas to nonauthentic production areas. However, the quality changes in LJ after plant production remained unclear. Objective: The objective of the study is to analyze the bioactive compounds, the corresponding gene expressions, and the endogenous plant hormone changes after 3-year introduction. Materials and Methods: In September 2014, LJ cuttings were planted in the two sites. In May 2017, LJ flower buds were randomly harvested and divided into three groups for liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), quantitative real-time reverse transcription-polymerase chain reaction, and indirect enzyme-linked immunosorbent assay, respectively. Results: The results showed that chlorogenic acid significantly decreased while luteoloside and luteolin significantly increased after plant introduction. Gene expression profiles related to biosynthesis pathways showed differentially expressed patterns although most of them were not significantly different. The down-regulated expression of hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase genes resulted in a decrease of chlorogenic acid content while the up-regulated expression of key genes related to synthesizing the universal precursor p-Coumaroyl-CoA and the branched luteoloside biosynthetic pathway led to an increase of luteoloside content. Altered endogenous phytohormone levels, especially jasmonic acid, regulate LJ quality changes. Conclusion: The data demonstrate that irrespective of the decrease of chlorogenic acid, the considerable increase of flavonoids demonstrated the benefit of LJ introduction. Abbreviations used: LJ: Lonicera japonica Thunb.; PAL: Phenylalanine ammonia-lyase (EC 4.3.1.24); C4H: Cinnamate 4-hydroxylase (EC 1.14.13.11); 4CL: 4-Hydroxycinnamoyl-CoA ligase/4-coumarate-CoA ligase (EC 6.2.1.12); HQT: Hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase (EC 2.3.1.99); CHS: Chalcone synthase (EC 2.3.1.74); CHI: Chalcone isomerase (EC 5.5.1.6); FNS: Flavone synthase (EC 1.14.11.22); GL: Glycosyltransferase (flavone 7-O-beta-glucosyltransferase, EC 2.4.1.81).
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Phytochemical analysis and cytotoxic activity of Cnidoscolus quercifolius Pohl (Euphorbiaceae) against prostate (PC3 and PC3-M) and breast (MCF-7) cancer cells p. 24
Raimundo G Oliveira Júnior, Christiane A A. Ferraz, Emanuella C V. Pereira, Pedrita A Sampaio, Maria Francilene S. Silva, Cláudia O Pessoa, Larissa A Rolim, Jackson Roberto Guedes da Silva Almeida
DOI:10.4103/pm.pm_6_18  
Background: Cnidoscolus quercifolius is a Brazilian medicinal plant often found in the Caatinga biome. Previous studies have described several pharmacological properties for this plant, including antiproliferative effect. However, there are still few pharmacological and phytochemical reports involving this plant. Objective: In this report, it was described the cytotoxic effect of extract and fractions obtained from the leaves of C. quercifolius. It was also reported for the first time the identification of two flavonoids in this species. Materials and Methods: Ethanol extract (EE) and fractions hexane, chloroform-Fr, ethyl acetate (AcOEt-Fr) and methanol (MeOH-Fr) were evaluated against prostate (PC3 and PC3-M) and breast (MCF-7) cancer cell lines. A preliminary phytochemical analysis was performed by thin layer chromatographic, while the content of total phenolic compounds and flavonoids was determined by colorimetric assays. EE and bioactive fraction (AcOEt-Fr) were selected for analysis by high-performance liquid chromatography-diode-array detector (HPLC-DAD). Results: Phytochemical analysis revealed that the samples were positive for the presence of several classes of secondary metabolites, mainly phenolic derivatives and flavonoids. EE and AcOEt-Fr presented the highest phenolic and flavonoid content. HPLC-DAD analysis of EE and AcOEt-Fr allowed the identification of two flavonoids (rutin and apigenin) not yet described for this species. Concerning the cytotoxicity evaluation, only AcOEt-Fr demonstrated a strong cytotoxic effect against all cell lines, presenting the half maximal inhibitory concentration values between 15.75 and 46.97 μg/ml. Conclusion: The results suggest that flavonoids may play an important role in the cytotoxic effect observed for this species. In addition, this report contributed to the phytochemical knowledge of the species through the identification and quantification of flavonoids. Abbreviations used: AcOEt-Fr: Ethyl acetate fraction; CHCl3-Fr: Chloroform fraction; EE: Ethanol extract; Hex-Fr: Hexane fraction; IC50: Half maximal inhibitory concentration; MeOH-Fr: Methanol fraction; TLC: Thin layer chromatography; UV: Ultraviolet.
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Comparative research of the curative effects of pinelliae rhizoma and pinelliae rhizoma praeparatum cum alumine on ovalbumin-induced allergic asthma in rats p. 29
Wei Peng, Da-Neng Wei, Yu-Jie Liu, Meng-Meng Zhang, Mei-Bian Hu, Chun-Jie Wu
DOI:10.4103/pm.pm_397_18  
Aim: This study was designed to compare the curative effects of Pinelliae Rhizoma (RPT) and Pinelliae Rhizoma Praeparatum cum Alumine (QPT) on ovalbumin (OVA)-induced allergic asthma in rats. Materials and Methods: Asthmatic rats were prepared by intraperitoneal injection and inhalation of OVA, and RPT and QPT at the doses of 120 and 240 mg/kg/day were administered orally for consecutive 7 days. Then, histopathological changes of the lung tissues were examined; levels of interleukin (IL)-4 and interferon (IFN)-γ in the bronchoalveolar lavage fluids (BALFs) and immunoglobulin (Ig) E in the serum were measured by enzyme-linked immunosorbent assay; eosinophils (EOS) in BALFs were also determined. Furthermore, the expressions of mucin 5AC (MUC5AC) and AQP-5 in the lung tissues were determined by quantitative real-time fluorogenic polymerase chain reaction and western blot assays. Besides, high-performance liquid chromatography-LTQ-Orbitrap-Elite-MS analysis was applied to compare the constituent's difference between the RPT and QPT. Results: The present results revealed that both the RPT and QPT have the potential curative effects for treating OVA-induced asthma in rats via inhibiting inflammatory reactions (suppressing EOS influx, IgE secretion, and release of IL-4 whereas increasing the release of IFN-γ) and modulating the mRNA and protein expressions of MUC5AC and AQP-5. In addition, our results also evidenced that QPT exhibited a better curative effect on allergic asthmatic rats compared to RPT. Conclusion: Our results are beneficial for confirming the traditional Chinese medicinal theory of “QPT is good at removing dampness to reduce phlegm.” Abbreviations used: BALF: Bronchoalveolar lavage fluid; ELISA: Enzyme-linked immunosorbent assay; EOS: Eosinophils; IFN: Interferon; IL: Interleukin; OVA: Ovalbumin; qRT-PCR: Quantitative real-time fluorogenic polymerase chain reaction; QPT: Rhizoma Pinelliae Praeparatum cum Alumine; RIPA: Radioimmunoprecipitation assay; RPT: Pinelliae Rhizoma; SDS-PAGE: Sodium dodecyl sulfate-polyacrylamide gel electrophoresis; TCM: Traditional Chinese medicine.
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Dietary oils ameliorate aluminum chloride-induced memory deficit in wistar rats p. 36
Ayush Kumar, Sanchari Basu Mallik, Samita Rijal, Nilanjan Changdar, Jayesh Mudgal, Rekha R Shenoy
DOI:10.4103/pm.pm_306_18  
Background: Alzheimer's disease (AD) is a progressive neurodegenerative disorder of the brain and is the most common form of dementia. There is no known cure for AD, and therefore, alternate therapies are warranted. Multiple reports state that nutritional or dietary interventions may have therapeutic potential and particularly associating high intake of n-3 polyunsaturated fatty acids to reduce risk for AD. In this direction, we have attempted to study the neuroprotective effects of dietary supplementation with canola oil, olive oil and rice bran oil in an aluminum chloride (AlCl3)-induced dementia model in Wistar rats. Materials and Methods: In the present study, spatial memory of AlCl3-treated animals, given supplementation with dietary oils, was evaluated using the Morris water maze. In addition, brain antioxidant parameters, acetylcholinesterase activity and serum biochemical markers were evaluated at the end of the study. Results: All dietary oils used in the present study showed ameliorative effect on the decline in spatial memory due to AlCl3treatment. However, among the three oils, canola oil exerted the most protective effect toward AlCl3-induced neuronal damage by improving spatial memory and brain antioxidant status. Apart from serum glucose, none of the biochemical parameters were altered by AlCl3or by dietary oil supplementation. Conclusion: The study highlights the protective effect of dietary oils rich in unsaturated fats against Alzheimer's dementia. Canola oil showed highest neuroprotective activity against AlCl3-induced dementia in rats. Abbreviation used: AD: Alzheimer's disease; ALA: Alpha-linoleic acid; DHA: Docosahexaenoic acid; PUFA: Polyunsaturated fatty acids; MWM: Morris water maze; AchE: Acetylcholinesterase; UFAW: University Federation for Animal Welfare.
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Ameliorative effect of morin, a plant flavonoid against Freund's complete adjuvant-induced polyarthritis in rats p. 43
Guozhi Zhang, Amit D Kandhare, Anwesha A Mukherjee, Subhash L Bodhankar, Hailei Yin
DOI:10.4103/pm.pm_351_18  
Background: Rheumatoid arthritis is a chronic relapsing autoimmune disorder with multifactorial etiology and prognosis. Morin [2-(2,4-dihydroxyphenyl)-3,5,7-trihydroxy-4H-1-benzopyran-4-one], a plant flavonoid reported having antioxidant and anti-inflammatory potential. Objective: The aim is to study the anti-arthritic activity of the morin against adjuvant-induced polyarthritis in rats. Materials and Methods: Polyarthritis was induced in female Wistar rats (150–180 g) using Freund's complete adjuvant (FCA) in the tail. Leflunomide (10 mg/kg, as a standard) and morin (10, 30 and 100 mg/kg) were administered orally daily for 28 days. Results: Treatment with morin (30 and 100 mg/kg) showed statistically significant inhibition (P < 0.05) in FCA-induced decrease in thermal hyperalgesia and mechanical hyperalgesia. It also showed significant attenuation (P < 0.05) in FCA-induced alterations in hematological parameters, aspartate aminotransferase, alanine transaminase, alkaline phosphatase, serum turbidity, albumin, C-reactive protein, blood sugar levels, and erythrocyte sedimentation rate. Altered levels of serum and liver lipid profile were significantly attenuated (P < 0.05) by morin (30 and 100 mg/kg). Morin treatment significantly decreased (P < 0.05) serum oxido-nitrosative stress, tumor necrosis factor-alpha (TNF-α), and interleukin-1β (IL-1β) levels. Morin (30 and 100 mg/kg) also significantly (P < 0.05) inhibited FCA-induced up-regulated liver TNF-α, IL-1β, IL-6, heme oxygenase-1, transforming growth factor-beta and down-regulated nuclear factor erythroid 2-related factor-2 messenger RNA expression. Histopathology alteration-induced by FCA was also reduced by morin treatment. Conclusion: Morin showed promising curative properties against FCA-induced polyarthritis in rats via modulation of endogenous biomarkers. Thus, morin can be considered as an alternative treatment regimen for the management of arthritis. Abbreviations used: AIA: Adjuvant-induced arthritis, ALT: Alanine transaminase, ALP: Alkaline phosphatase, AST: Aspartate aminotransferase, BSL: Blood glucose level, CRP: C-reactive protein, ESR: Erythrocyte sedimentation rate, FCA: Freund's complete adjuvant, HO: Heme oxygenase, HDL: High-density lipoprotein, IL-1β: Interleukin-1 beta, LDL: Low-density lipoprotein, MDA: Malondialdehyde, NO: Nitric oxide, Nrf2: Nuclear factor-like 2, ROS: Reactive oxygen species, GSH: Reduced glutathione, RT-PCR: Reverse transcription-polymerase chain reaction, SOD: Superoxide dismutase, TC: Total cholesterol, TGF-β: Transforming growth factor-beta, TG: Triglyceride, TNF-α: Tumor necrosis factor-alpha, VLDL: Very low-density lipoprotein.
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Durio zibethinus rind extract mediated green synthesis of silver nanoparticles: Characterization and biomedical applications p. 52
Samuggam Sumitha, Sethu Vasanthi, Sivadasan Shalini, Suresh V Chinni, Subash C B. Gopinath, Sathasivam Kathiresan, Periasamy Anbu, Veerasamy Ravichandran
DOI:10.4103/pm.pm_400_18  
Background: Silver nanoparticles play a profound role in the field of biology and medicine due to its attractive physiochemical properties. Objective: The present work was aimed to develop green approach for the synthesis of silver nanoparticles (DRAgNPs) using D. zibethinus rind aqueous extract and determination of its antimicrobial and cytotoxic effect against brine shrimp. Materials and Methods: Aqueous extract of D. zibethinus rind was used to reduce silver nitrate to silver nanoparticles. The various reaction parameters were optimized, and DRAgNPs were characterized for size, shape, and nature. Results: Surface plasmon resonance confirmed the formation of DRAgNP's with maximum absorbance at λmaxof 418 nm. Scanning transmission electron microscopy and transmission electron microscope images revealed the morphology of the DRAgNPs were spherical with size range of 20 and 60 nm. Atomic force microscopy images confirmed the average particles size of DRAgNPs was to be 55 nm. The stability of the nanoparticles was also confirmed by the zeta potential which was found to be −15.82 mV. X-ray powder diffraction and energy-dispersive X-ray spectroscopy analysis confirmed the nature and the presence of Ag. DRAgNPs showed considerable antimicrobial activity against Salmonella typhimurium, Escherichia coli, Salmonella typhi, Staphylococcus aureus, Staphylococcus haemolyticus, and Bacillus subtilis, and exhibited better cytotoxicity against brine shrimp (LC50 =2.55 mg/mL). Conclusion: Based on the present study, it can be concluded that the green synthesis of silver nanoparticles using D. zibethinus rind is an eco-friendly and inexpensive method, and DRAgNPs could be used in the field of water treatment, pharmaceuticals, biomedicine, biosensor, and nanotechnology in the near future. Abbreviation used: SEM: Scanning transmission electron microscopy; TEM: Transmission electron microscope; AFM: Atomic force microscopy; XRD: X-ray powder diffraction; EDX: Energy-dispersive X-ray spectroscopy.
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Vasorelaxant effect of the ethanol extract from Valeriana fauriei briquet root and rhizome on rat thoracic aorta p. 59
Bumjung Kim, Shin-Saeng Ma, Cheolmin Jo, Somin Lee, Hanna Choi, Kyungjin Lee, Inhye Ham, Ho-Young Choi
DOI:10.4103/pm.pm_152_18  
Background: Valeriana fauriei has been used to treat anxiety and cardiovascular disease in many countries; however, the mechanisms of the vasorelaxant effect of V. fauriei roots and rhizomes have not been elucidated. Purpose of the Study: Considering the increased use of traditional medicines for treating hypertension, the aim of this study was to examine the vasorelaxant effect and investigate the mechanisms of V. fauriei root and rhizome. Materials and Methods: Using an isolated organ-chamber technique, the vasorelaxant effects of 70% ethanol extract of V. fauriei root and rhizome (VFE) were tested in rat aortic ring segments. Results: VFE concentration dependently relaxed both endothelium-intact and endothelium-denuded aortic rings that were precontracted by phenylephrine (PE) or potassium chloride (KCl). VFE relaxed the aortic rings precontracted by PE (1 μM) irrespective of K+ channel blocker pretreatment such as tetraethylammonium, glibenclamide, or 4-aminopyridine. VFE inhibited vasoconstriction caused by extracellular Ca2+ influx through receptor-operative Ca2+ channels (ROCCs) or voltage-dependent Ca2+ channels (VDCCs) activated by PE or KCl, respectively, in Ca2+-free Krebs–Henseleit buffer. Conclusion: These results suggested that the vasorelaxant effects of VFE were endothelium independent and not related to K+ channels, such as Ca2+-activated K+ channels, ATP-sensitive K+ channels, and Voltage-gated K+ channels. In addition, VFE relaxed the aortic rings by blocking the influx of extracellular Ca2+ through ROCCs and VDCCs. Abbreviations used: ACE: Angiotensin-converting enzyme; VFE: The 70% EtOH extract of Valeriana fauriei root and rhizome; KH: Krebs–Henseleit; PE: Phenylephrine; KCl: Potassium chloride; TEA: Tetraethylammonium; 4-AP: 4-Aminopyridine; CaCl2: Calcium chloride; EGTA: Ethylene glycol-bis (2-aminoethylether)-N, N, N′, N′-tetraacetic acid; DMSO: Dimethyl sulfoxide; ROCCs: Receptor-operative Ca2+ channels; VDCCs: Voltage-dependent Ca2+ channels; KIR: Inward-rectifier K+ channels; KATP: ATP-sensitive K+ channels; KV: Voltage-gated K+ channels; KCa: Ca2+-activated K+ channels.
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Toxicity study of silver nanoparticles synthesized from aqueous flower extract of Scrophularia striata on MCF-7 human breast cancer cell line p. 66
Roohangiz Mameneh, Massoumeh Shafiei, Ali Aidy, Elahe Karimi, Behzad Badakhsh, Naser Abbasi
DOI:10.4103/pm.pm_78_18  
Background: Silver nanoparticles (AgNPs) have been used as an antimicrobial and disinfectant agent. Nevertheless, there is limited data about antitumor potential. This study has focused on investigating cytotoxic effects of AgNPs from Scrophularia striata flower extract on MCF-7 breast cancer cells and its mechanism of action. Materials and Methods: Thus, a green method was created for the synthesis of AgNPs using an aqueous extract of S. striata flower. Synthesis of AgNPs was described by different analytical techniques including ultraviolet-visible spectrophotometer, field-emission scanning electron microscopy, X-ray diffraction, and Fourier transforms infrared spectroscopy. Cell viability was determined by the 3-[4, 5-dimethylthiazol-2-yl]-a 2,5-diphenyltetrazolium bromide assay. Reactive oxygen species (ROS) formation was measured using probe 2', 7'-dichlorofluorescein diacetate and intracellular calcium (Cai2+) was evaluated with probe flu3-AM. Cells were treated with different concentrations of AgNPs (1, 3, 6, 10, 15, 25, 50, and 100 μg/mL). Results: The results showed that AgNPs hindered cell growth in a dose-dependent manner. AgNPs appeared to have dose-dependent cytotoxicity against MCF-7 cells through activation of the ROS generation and an increase in the intracellular Cai2+ (IC50 52 ± 3.14). Conclusion: In conclusion, the results of this preliminary study demonstrated that AgNPs from S. striata flower extract may be a potential therapeutic agent for human breast cancer treatment. Abbreviations used: S. striata: Scrophularia striata; FESEM: Field-emission scanning electron microscopy; XRD: X-ray diffraction; FTIR: Fourier transforms infrared spectroscopy; MTT: 3-[4, 5-dimethylthiazol-2-yl]-a 2,5-diphenyltetrazolium bromide; ROS: Reactive oxygen species; DCF: 2′, 7′-dichlorofluorescein diacetate; Cai2+: Intracellular calcium.
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New formulation containing curcuminoids: Method validation and dissolution profile p. 73
Nayara Luiza Oliveira Ferreira, Mariana Cristina Morais Rodrigues, Rejanne Lima Arruda, Pierre Alexandre Santos, Maria Teresa Freitas Bara, Edemilson Cardoso Conceição
DOI:10.4103/pm.pm_50_18  
Background: Curcuma longa, popularly known as turmeric, contains secondary metabolites called curcuminoids (curcumin, demethoxycurcumin, and bisdemethoxycurcumin) which are responsible for its therapeutic activity. Objective: The aim of this work was to obtain capsules using C. longa soft extract, to develop and validate a method for quantification of curcuminoids, and to study the dissolution profile of the capsules. Materials and Methods: The liquid extract was obtained through the percolation method and concentrated until it is characterized as soft extract. A high-performance liquid chromatography (HPLC) method was developed and validated for quantification of curcuminoids in the soft extract. The granule was obtained through wet granulation process and was encapsulated and the method for quantification of the markers was covalidated. Results: The soft extract showed 3.45% of bisdemethoxycurcumin, 3.03% of demethoxycurcumin, and 14.83% of curcumin, corresponding to 21.31% of total curcuminoids. The capsules had an average weight of 420 mg with approximately 21.56 mg of curcuminoids. The capsules showed better dissolution profile in neutral medium that simulates the pH of duodenum; however, in the stability test, only the variation found for bisdemethoxycurcumin was in compliance with the permitted. Conclusions: A soft extract of C. longa standardized in the three main curcuminoids was produced, as well as hard gelatinous capsules containing a dose of 21.56 mg of total curcuminoids. Thus, a new phytotherapeutic formulation was proposed, which proves to be an interesting strategy to carry out the active principles of C. longa. Abbreviations used: HPLC: High-performance liquid chromatograph; LOD: Limit of detection; LOQ: Limit of quantification; PDA: Photodiode array detector; PTFE: Hydrophobic polytetrafluoroethylene; TLC: Thin-layer chromatography; UV: Ultraviolet
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Determination of isoorientin levels in rat plasma by ultra-high performance liquid chromatography coupled with diode array detector and its application to a pharmacokinetic study p. 81
Doan Nguyen Kieu Trang, Yu Chul Kim, Seung-Hwan Kwon, Choon-Gon Jang, Han-Joo Maeng
DOI:10.4103/pm.pm_68_18  
Background: Isoorientin is a C-glycosylflavone and a pharmacologically active ingredient found in various medicinal plants. Objective: The aim of this study is to develop a specific and reproducible ultra-high-performance liquid chromatography-diode array detector (UHPLC-DAD) based method to quantify isoorientin in rat plasma and to apply the devised method to a pharmacokinetic study in rats. Materials and Methods: Simple protein precipitation with methanol was utilized to extract isoorientin and rutin (the internal standard) from rat plasma. Analytes were separated on an UHPLC Phenomenex Luna Omega Polar C18 column (100 mm × 2.1 mm, 1.6 μm) by gradient elution using a mobile phase containing 1% aqueous acetic acid and 100% acetonitrile at the flow rate of 0.25 mL/min. Results: The developed UHPLC-DAD method showed good linearity (R2 = 0.9993) over the concentration range 20–5000 ng/mL with a lower limit of quantification of 20 ng/mL. Intra- and inter-day precisions were <10.7% and accuracy were within the range of 89.4%–101.3%. The recovery of isoorientin from plasma was in acceptable range (89.4%–95.5%). The devised method was successfully applied to a pharmacokinetic study after a single oral administration of isoorientin to rats. Conclusion: This is the first report of a simple, rapid, and cost-effective UHPLC-DAD-based method for quantifying isoorientin in rat plasma and its application to a pharmacokinetic study. Abbreviations used: UHPLC: Ultra-high-performance liquid chromatography; DAD: Diode array detector; LC-MS/MS: Liquid chromatography-tandem mass spectrometry; LLOQ: Lower limit of quantification; IS: Internal standard; QC: Quality control; RSD: Relative standard deviation; Cmax: The maximum plasma concentration; Tmax: The time point to reach the maximum concentration; AUClast: The area under the curve from time zero to the last measurable point, AUCinf: The area under the curve from time zero to infinity; ke: Terminal elimination rate constant; t1/2: Terminal half-life; CL/F: Oral clearance; Vd/F: Apparent volume of distribution after oral administration; SD: Standard deviation; SPE: Solid-phase extraction.
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Clinacanthus nutans induced reactive oxygen species-dependent apoptosis and autophagy in HCT116 human colorectal cancer cells p. 87
Kar Suen Wang, Chim Kei Chan, Ahmad Fadhlurrahman Ahmad Hidayat, Yau Hsiung Wong, Habsah Abdul Kadir
DOI:10.4103/pm.pm_299_17  
Background: Clinacanthus nutans (Burm.f.) Lindau is a medicinal herb that is conventionally used for the treatment of skin rashes, insect bites, snake bites, diabetes, and cancer. Objective: Our study aims to investigate the apoptosis- and autophagy-inducing effects of C. nutans in HCT116 human colorectal cancer cells. Materials and Methods: Cytotoxicity of ethanol extract, hexane, ethyl acetate, and aqueous fractions of C. nutans against various cancer cell lines was determined via MTT assay. Apoptosis assays including annexin V, mito-ID, and Hoechst 33342/propidium iodide staining were carried out. The level of intracellular reactive oxidative species was determined using flow cytometry. Western blot analysis was carried out to assess the protein expression in C. nutans ethyl acetate fraction (CNEAF)-treated HCT116 cells. Results: CNEAF was found to exert the strongest cytotoxic effect against HCT116 cells (IC50 =48.81 ± 1.44 μg/mL). CNEAF-induced apoptosis was evidenced by nuclear morphological alterations, phosphatidylserine externalization, dissipation of mitochondrial membrane potential, and elevation of intracellular reactive oxygen species (ROS) level. Dissipation of mitochondrial membrane potential was attributed to the upregulation of Bax and Bak accompanied by downregulation of Bcl-2 and Bcl-xL, leading to caspase-3, -9, -8, and -10 activation. Interestingly, an upregulation of death receptor 5 was detected, suggesting involvement of intrinsic and extrinsic pathways. In addition, the occurrence of autophagy by CNEAF was supported by LC-3 accumulation and p62 degradation. The reduction of intracellular ROS level by N-acetylcysteine showed that the apoptosis and autophagy induced by CNEAF is ROS dependent. Conclusions: CNEAF induced ROS-dependent apoptosis and autophagy on HCT116 cells. Abbreviations used: CNEAF: Clinacanthus nutans ethyl acetate fraction; ROS: Reactive oxygen species; PS: Phosphatidylserine; NAC:N-Acetyl Cysteine; Bax: Bcl-2-associated X; Bcl-2: B-cell lymphoma 2; DR-5: Death receptor 5 Bak: Bcl-2-antagonist/killer 1; Bcl-xL: B-cell lymphoma-extra large.
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Pharmacological mechanisms of the water leaves extract of Lysiphyllum strychnifolium for its anti-inflammatory and anti-hyperuricemic actions for gout treatment p. 98
Vilasinee Hirunpanich Sato, Savita Chewchinda, Narawat Nuamnaichati, Supachoke Mangmool, Bunleu Sungthong, Pilanthana Lertsatitthanakorn, Shinako Ohta, Hitoshi Sato
DOI:10.4103/pm.pm_14_18  
Background: There have been anecdotal reports from Thai hyperuricemic patients that the leaves of Lysiphyllum strychnifolium could reduce plasma uric acid level and relieve inflammation of gout. However, no research to support these effects has been conducted. Objectives: This study was aimed to evaluate the anti-inflammatory and hypouricemic effects of L. strychnifolium leaves extract and to investigate the pharmacological mechanisms of these effects. Materials and Methods: The anti-inflammatory effect of L. strychnifolium was evaluated in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Effects of L. strychnifolium on xanthine oxidase (XO) were examined in vitro and in vivo using potassium oxonate (PO)-induced hyperuricemic mice. In addition, the antioxidant activity of L. strychnifolium was determined. Results: L. strychnifolium significantly reduced the mRNA expression of cyclooxygenase-II, inducible nitric oxide synthase, transforming growth factor-β, and tumor necrosis factor-α in LPS-stimulated RAW 264.7 cells (P < 0.05). It exhibited a noncompetitive inhibition of XO activity with IC50and K i of 231 μg/ml and 177 μg/ml, respectively. Oral administration of L. strychnifolium (100 and 200 mg/kg) significantly lowered the plasma uric concentration (P < 0.05) in PO-induced hyperuricemic mice and inhibited 56.9% and 66.3% of the hepatic XO activity, respectively, compared to control hyperuricemic mice (P < 0.05). L. strychnifolium did not significantly decrease the protein expression of solute carrier family 22 member 12 in the renal cortex. Total phenolic and flavonoid contents were determined to be 197.8 ± 5.8 mg gallic acid equivalence/g extract and 32.2 ± 1.2 mg quercetin equivalent/g extract, respectively. The IC50of the inhibition of 2,2-diphenyl-1-picrylhydrazyl radical was 44.32 μg/ml. Conclusion: The present study first provided scientific evidence for the anti-inflammatory, anti-hyperuricemic and antioxidant effects of L. strychnifolium leaves extract in vitro and in vivo, suggesting the possibility of this plant to treat gout. Abbreviations used: ABTS: 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid); COX-II: Cyclooxygenase-II; DMEM: Dulbecco's modified Eagle medium; DPPH: 2,2-Diphenyl-1-picrylhydrazyl; FBS: Fetal bovine serum; FRAP: Ferric reducing antioxidant power; GAE: Gallic acid equivalence; GLUT9: Glucose transporter 9; iNOS: Inducible nitric oxide synthase; LPS: Lipopolysaccharide; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; PO: Potassium oxonate; P/S: Penicillin/streptomycin; QE: Quercetin equivalent; SLC22A12: Solute carrier family 22 member 12; TNF-α: Tumor necrosis factor-α; TGF-β: Transforming growth factor-β; URAT: Urate-anion transporter 1; XO: Xanthine oxidase.
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Exogenous intervention with sodium hydrosulfite enhances the quality of Radix Scutellariae by modulating antioxidases and secondary metabolites p. 107
Song Qi, Gao Huiru, Wang Jiahui, Zhang Aihua, Meng Xiangcai
DOI:10.4103/pm.pm_598_17  
Introduction: The transformation of wild herbal medicine into cultivation results in decreased quality, becoming an issue of concern in China, even has a word, “traditional Chinese medical science would be destroyed by herbal medicine.” Now, there is no way to acquire superior herb medicine. Reactive oxygen species (ROS) are generated under stress and represent key plant products that induce secondary metabolism. ROS may be important intermediates that link environmental stress to secondary metabolism. Sodium hydrosulfite (Na2S2O4) is just one compound that produces O2•-. Materials and Methods: The fresh roots of Scutellaria baicalensis were treated with 0.0004, 0.04, and 4 mmol/L Na2S2O4, respectively. A stress model was established to elucidate the change of secondary metabolism, antioxidant enzyme system, and enzymes relating to flavonoids. Results: Na2S2O4regulated the activities of β-glucuronidase (GUS) and UDP-glucuronate baicalein 7-O-glucuronosyltransferase (UBGAT) and promoted the transformation of flavonoid glycosides into aglycones with greater antioxidant capacities. In fresh S. baicalensis roots, Na2S2O4decreased the activities of superoxide dismutase, catalase, and peroxidase (POD), which led to a substantial accumulation of hydrogen peroxide (H2O2). H2O2increased the expression of phenylalanine ammonia lyase, GUS, and UBGAT genes and promoted the synthesis and transformation of flavonoids. Moreover, polyphenol oxidase regulated the flavonoid content to maintain the oxidation balance. Although the content of flavonoid glycosides was slightly reduced, the total flavonoid content remained high. Notably, the content of baicalein, an antioxidant compound, increased from 0.28% to 1.96%. Conclusions: ROS facilitated the dominant role of secondary metabolites and enhanced the quality of Radix Scutellariae. Abbreviations used: APX: Ascorbate peroxidase, CAT: Catalase, GUS: β-glucuronidase, PAL: Phenylalanine ammonia lyase, POD: Peroxidase, PPO: Polyphenol oxidase, ROS: Reactive oxygen species, SOD: Superoxide dismutase, UBGAT: UDP-glucuronate baicalein 7-O-glucuronosyltransferase.
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An ethnobotanical and ethnomedicinal survey of traditionally used medicinal plants in Seymour, South Africa: An attempt toward digitization and preservation of ethnic knowledge p. 115
Lisa Valencia Buwa-Komoren, Buyisile Mayekiso, Zoleka Mhinana, Ariyo Lateef Adeniran
DOI:10.4103/pm.pm_259_18  
Introduction: Medicinal plants have been used for the treatment of both infectious and noninfectious diseases by the majority of the world's population for many years. The low socioeconomic standing of Eastern Cape population suggests that the majority of people use traditional methods of health care. Many of the rural communities in this province have no access to Western medical health care and rely on traditional medicine for their primary health-care needs. Materials and Methods: An ethnobotanical survey was conducted from February 2013 to December 2015 to investigate the use of medicinal plants by the people of Seymour in the Eastern Cape Province of South Africa. Information was gathered from nine traditional healers, seven herbalists, and 18 elderly villagers. Results: The information collected revealed six ailment categories that were treated with a wide range of medicinal plants. A total of 50 plant species belonging to 29 families were reported to be used in the treatment of various ailments. Members of the family Asteraceae, Euphorbiaceae, Fabaceae, and Rutaceae had the highest number of species used in traditional healing. Leaves were reported to be the most frequently used plant part, followed by roots, bark, stem, and then corms and rhizomes. The survey indicated that the most prominent method of herbal administration used is orally via extracts that were obtained by boiling, either as a decoction or concoction. Conclusions: This study has documented important information on medicinal plants used by people, traditional healers, and herbalists of Seymour region in the Eastern Cape Province of South Africa to treat various ailments. Abbreviations used: ACE: Angiotensin-converting enzyme; CNS: Central nervous system; ENT: Ear, nose and throat; HIV: Human Immunodeficiency virus; ICF: Informant consensus factor; Nur: Total number of use report; Nt: Number of taxa; TB: Tuberculosis
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Total anthocyanins and cyanidin-3-O-glucoside contents and antioxidant activities of purified extracts from eight different pigmented plants p. 124
Luo Yang, Cai Rong-Rong, Fang Ji-Li, Yuan Ke
DOI:10.4103/pm.pm_162_18  
Background: Anthocyanin, a subcategory of flavonoid, is a natural water-soluble pigment. There are many plants rich in anthocyanins, and a high intake of anthocyanin food has been shown to have potential beneficial effects on various chronic diseases. Objective: The objective was to evaluate the contents of total anthocyanin and cyanidin-3-O-glucoside (C-3-G) and their contribution to the antioxidant activities in commonly consumed pigmented plants. Materials and Methods: The total anthocyanin in Lonicera caerulea L., Rubus fruticosus L., Ribes nigrum L., Morus alba L., Zea mays L. seed, Z. mays L. cob, Brassica oleracea L., and Dioscorea alata L. was extracted by tissue-smashing extraction method, and then the contents of total anthocyanin (TAC) and C-3-G contents (C-3-GC) in the purified extracts were determined by pH differential method and high-performance liquid chromatography, respectively. Antioxidant activities were assessed by 2,2-diphenyl-1-picrylhydrazyl, ferric-reducing antioxidant power, and total reducing power (TRP) assays. Results: TAC ranged from 97.11 to 320.27 milligrams cyanidin-3-glucoside equivalents per Gram of dry weight. TAC, C-3-GC, and antioxidant activities in most berry extracts were higher than that in vegetables. Z. mays cob showed the similar TAC, C-3-GC, and antioxidant activities to L. caerulea, and the two vegetables were the lowest. The major anthocyanin in the berries and grains was identified as C-3-G. There was a significant positive correlation between antioxidant activity and TAC. Conclusions: The closer the plant color is to black, the higher TAC is, and the stronger its antioxidant activity is. Z. mays cob will be a promising source of anthocyanin. This study provides a theoretical basis for the use of anthocyanin in functional food and further pharmacological research. Abbreviations used: C-3-G: cyanidin-3-O-glucoside; C3GC: cyanidin-3-O-glucoside contents; DPPH: 2, 2-diphenyl-1-picrylhydrazyl; FRAP: Ferric-reducing antioxidant power; HPLC: High-performance liquid chromatography; TAC: Total anthocyanin content; TAE: Total anthocyanin extract; TE: Trolox equivalent; TRP: Total reducing power; TPTZ: 1,3,5-tri (2-pyridyl)-2,4,6-triazine; TSE: Tissue-smashing extraction.
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Immunomodulatory activity of an Indonesian herbal formulation for respiratory disorder p. 130
Zullies Ikawati, Triana Hertiani, Fauzia Izzati
DOI:10.4103/pm.pm_314_18  
Objective: The aim of this study is to determine the activity of an Indonesian herbal formulation, which is specific for respiratory disorders and contains a combination of Curcuma xanthorrhiza rhizome, Vitex trifolia leaves, Zingiber officinale rhizome, Citrus fruit, and Echinacea purpurea herb, as an immunomodulator in mice. Materials and Methods: Thirty male Balb/c mice were divided into five groups as follows: negative control, E. purpurea for reference, and three doses of herbal formula. The samples were administered daily for 14 days, and immune response was induced by hepatitis B vaccine on days 7 and 14. On day 18, blood was collected via the retro-orbital plexus. Macrophage phagocytosis and lymphocyte proliferation assay and measurement of immunoglobulin G (IgG) antibody titers were carried out on day 20. Results: The herbal formula can significantly increase the phagocytic index compared with the negative control. Interestingly, the herbal formula showed higher activity compared with E. purpurea-only treatment. This indicates the possible synergistic effects of the herbal components in the formula. This synergistic pattern was also observed in the production of IgG antibody. The herbal formula can significantly increase IgG titer compared with the negative control group (P < 0.05). However, the increase was nonsignificant compared with E. purpurea-only treatment. On the other hand, the herbal formula showed less effect on the induction of lymphocyte proliferation compared with E. purpurea-only treatment. Low-dose treatments showed suppressive effects. Conclusions: The herbal formula showed more immunostimulatory activity on nonspecific responses than specific immune ones. Other herbal components may contribute to the overall immunomodulatory effect of the studied herbal formula. Abbreviations used: BW: Body weight; CMC: Cell-mediated cytotoxicity; CMC-Na: Carboxymethyl chitosan in Na-salt form; ELISA: Enzyme-linked immunosorbent; HBsAg: Hepatitis B surface antigen; IgG: Immunoglobulin G; OD: Optical density; PBS: Phosphate-buffered saline; RPMI: Roswell park memorial institute; PI: Phagocytic index; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.
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Comparative studies of selected Calophyllum plants for their anti-inflammatory properties p. 135
Siau Hui Mah, Soek Sin Teh, Gwendoline Cheng Lian Ee
DOI:10.4103/pm.pm_212_18  
Background: Calophyllum (Clusiaceae) plants have been used as traditional medicine for rheumatism, vein problems, hemorrhoids, and gastric ulcers. The traditional uses of this genus prompted us to investigate six Malaysian Calophyllum species which are Calophyllum inophyllum, Calophyllum soulattri, Calophyllum lowii, Calophyllum teysmannii, Calophyllum benjaminum and Calophyllum javanicum for their anti-inflammatory properties. Materials and Methods: The stem bark of six plants was extracted with n-hexane (Hex), ethyl acetate (EA), and methanol (MeOH). The activities of the extracts were evaluated by determining the inhibition of nitric oxide (NO) production by lipopolysaccharide-induced RAW 264.7 cells, as well as protein denaturation. Results and Discussion: The C. lowii extracts showed the most significant activities against NO production with IC50values of <40 μg/mL. For the protein denaturation test, C. teysmannii extracts showed the strongest effect with IC50values of <100 μg/mL. The results indicated that C. lowii and C. teysmannii are effective in both assays. Besides, the Hex extracts of these Calophyllum plants possess stronger activities than the EA and MeOH extracts. The anti-inflammatory properties are contributed by the secondary metabolites present in the crude extracts, specifically flavonoid, triterpenes, and xanthones. Conclusion: These results confirm the potential of Calophyllum plants to serve as lead agents in preventing inflammation. Abbreviations used: NO: Nitric oxide; spp.: Species; Hex: Hexane; EA: Ethyl acetate; MeOH: Methanol.
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Anti-inflammatory and anti-oxidative effects of Centella asiatica extract in lipopolysaccharide-stimulated BV2 microglial cells p. 140
Nootchanat Mairuae, Poonlarp Cheepsunthorn, Benjaporn Buranrat
DOI:10.4103/pm.pm_197_18  
Background: Neuroinflammation and oxidative stress mediated by microglial activation have been reported to play a critical role in the pathogenesis of neurodegenerative diseases. Therefore, inhibition of microglial activation using herbal medicine may be a potential candidate for the treatment of such diseases. Objective: The goal of this study was to investigate the anti-inflammatory and anti-oxidative effects of Centella asiatica extract (CA) on lipopolysaccharide (LPS)-stimulated BV2 microglial cells. Methods: BV2 microglial cells were treated with LPS in the presence or absence of CA extract. The levels of nitric oxide (NO), tumor necrosis factor-α (TNF-α) and reactive oxygen species (ROS) was measured using Griess reagent assay, enzyme-linked immunosorbent assay (ELISA) assay and CM-H2DCFDA, respectively. The nuclear levels of nuclear factor kappa B (NF-kB) p65 were detected using immunofluorescence and ELISA assay. Results: CA treatment resulted in significant and concentration-dependently reduced the LPS-induced production of NO, TNF-α, and ROS compared to the untreated group. CA treatment exerted an anti-inflammatory effect by suppressing NF-kB p65 translocation and the activation of Akt and the extracellular-signal-regulated kinase 1/2 (ERK1/2) pathway in LPS-stimulated BV2 cells. Conclusion: Taken together, these results show that CA exerts antioxidative activity by suppressing ROS production and that it exerts anti-inflammatory activity by suppressing LPS-induced NO and TNF-α production in BV2 microglial cells. These effects may occur through inhibition of Akt and the ERK1/2-mediated NF-kB pathway. The results presented here, coupled with traditional therapeutic claims, suggest that CA may be beneficial for treating neurodegenerative diseases mediated by microglial cells. Abbreviations used: AD: Alzheimer's disease; ALS: Amyotrophic lateral sclerosis; CA: Centella asiatica; DMSO: Dimethyl sulfoxide; DMEM: Dulbecco's modified Eagle's medium; ECL: Enhanced chemiluminescence; ELISA: Enzyme-Linked immunosorbent assay; ERK1/2: Extracellular-signal-regulated kinase 1/2; FBS: Fetal bovine serum; LPS: Lipopolysaccharide; MS: Multiple sclerosis; MSU: Mahasarakham University; NF-kB: Nuclear factor kappa B; NO: Nitric oxide; PD: Parkinson's disease; ROS: Reactive oxygen species; SD: Standard deviation; TNF-α: Tumor necrosis factor-α.
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Convenient preparation of 2″-O-rhamnosyl icariside II, a rare secondary flavonol glycoside, by recyclable and integrated biphase enzymatic hydrolysis p. 147
Chunlai Feng, Yi Lu, Yiying Zhou, Hui Pang, Yuping Shen, Huan Yang
DOI:10.4103/pm.pm_398_18  
Background: 2″-O-rhamnosyl icariside II, a rare secondary flavonol glycoside in Epimedii Folium (EF), has much better in vivo bioactivities than its original glycoside epimedin C. Its preparation methods, such as acidic hydrolysis, are of low efficiency, with by-products generated. Objective: The objective of this study was to establish a novel catalysis system for convenient preparation of this compound based on the recyclable and integrated biphase enzymatic hydrolysis. Materials and Methods: β-dextranase was selected from five commercial enzymes due to the best catalysis performance. After optimization of conditions, the biphase system was constructed with propyl acetate and HAc-NaAc buffer (pH 4.5) (3:2, v/v) containing β-dextranase/epimedin C (1:2, w/w), and the hydrolysis was performed at 60°C for 40 min. Results: Epimedin C was completely hydrolyzed to 2″-O-rhamnosyl icariside II, and 93.38% of the product has been transferred into organic phase; moreover, a high conversion rate had been achieved at 91.69% even after the enzyme solution was used for four cycles. In addition, the procedure was much simplified compared with conventional enzymatic hydrolysis. Conclusion: The newly proposed strategy is an efficient and promising approach for the preparation of 2″-O-rhamnosyl icariside II in industrial application. Abbreviations used: EF: Epimedii Folium; ACN: Acetonitrile; MeOH: Methanol; PTFE: Polytetrafluoroethylene; SD: Standard deviation; ILO-ICSC: International Labor Organization-The International Chemical Safety Cards; HMDB: Human Metabolome Database; HSDB: Hazardous Substances Data Bank.
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Optimization of ultrasonic-assisted extraction of bioactive compounds from Sargassum henslowianum using response surface methodology p. 156
Yongguang Bi, Yanshan Lu, Hong Yu, Lili Luo
DOI:10.4103/pm.pm_347_18  
Background: Sargassum henslowianum has become an important source for the food industry as well as medicinal applications. In recent years, varieties of bioactive components in S. henslowianum and its activities have reported. However, the optimized extraction conditions of polysaccharides and polyphenols in S. henslowianum was unknown, and their activities need to be explored more. Objective: This study was to optimize the ultrasound-assisted extraction of polysaccharides and polyphenols from S. henslowianum and test the biological activity of the extract. Materials and Methods: Response surface methodology was used to optimize extraction conditions, and the antioxidant activity of the extracts was evaluated by radical scavenging assay, α-Glucosidase inhibition, and cytotoxicity on MCF-7. Results: The optimal conditions of extracting polysaccharides were shown as following: ultrasonic time for 40 min, ultrasonic power for 330 W, solid-to-liquid ratio for 1:36 g/mL, the extraction yield reached 12.63% under above parameters. The optimum conditions of ultrasonic-assisted extraction of total polyphenols were as following: ultrasonic time for 102 min, ultrasonic power for 377 W, alcohol concentration for 62%, under these conditions, the extraction yield reached 11.45%. Besides, the extracts of polyphenols possessed stronger activity. Conclusion: This study provides the scientific guidance for further exploitation and utilization of S. henslowianum. Abbreviations used: RSM: Response Surface Methodology; BBD: Box-Behnken Design; ANOVA: Analysis of variance
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Metabolic profiling analysis of three processed rhizomes of Curcuma wenyujin Y.H. Chen et C. Ling by ultra-performance liquid chromatography/time-of-flight mass spectrometry p. 164
Min Hao, De Ji, Lin Li, Lianlin Su, Ji Zhang, Qiaohan Wang, Wei Gu, Chengxi Jiang, Tulin Lu, Chunqin Mao
DOI:10.4103/pm.pm_399_18  
Background: In China, rhizome of Curcuma wenyujin (CW) is used to improve blood stasis syndrome-related diseases for many years. Nonsteamed, steamed, and boiled with vinegar rhizomes of CW can be used as three different traditional Chinese medicine s, named as Pian-Jiang-Huang (PJH), Sheng-E-Zhu (SEZ), and Cu-E-Zhu (CEZ), respectively. After processing, the therapeutic effects have changed. Objective: In order to illustrate the effective substance of the three kinds of rhizome of CW, an ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS) method coupled with statistical approach was developed. Materials and Methods: Fresh rhizomes of CW were processed into PJH, SEZ, and CEZ according to Chinese pharmacopoeia 2015. UPLC-Q/TOF-MS coupled with chemometric analysis was applied to compare the differences in chemical profiles of the three kinds of rhizome of CW. Results: In this study, 21 compositions in the rhizome of CW were identified. The one-way ANOVA of relative intensities of the 21 compositions was processed by SPSS software version 20.0. The column superposed graph of the relative intensities of 21 components shows that the total relative intensities of the 21 components in SEZ are the topmost and that of PJH is about the same as CEZ, but the proportion of each component is quite differ from each sample. Principal component analysis and orthogonal partial least squares discrimination analysis were processed by Simca-p14.1 software. Finally, five ions including curcumenone, curcumol, curzerenone, furanodiene, and germacrone were discovered to be the Q-Makers of the three kinds of rhizome of CW. Conclusion: This study provides a reliable research basis for the clinical application of the three kinds of rhizome of CW. Abbreviations used: PJH: Pian-Jiang-Huang; SEZ: Sheng-Er-Zhu; CEZ: Cu-Er-Zhu; CW: Curcuma wenyujin; PCA: Principal component analysis; OPLS-DA: Orthogonal partial least squares discrimination analysis; RT: Retention time; TCM: Traditional Chinese medicine; TIC: Total ion chromatogram; QC: Quality control; RSD: Relative standard deviation.
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Protective effect of Schisandra chinensis extracts against H2O2-induced oxidative damage in RINm5F cells p. 172
Lulu Gong, Wei Dong, Xuechun Lu, Guangxin Yuan, Yunpeng Sun, Peige Du, Hui Yang, Meitong Pan, Yingnuo Li
DOI:10.4103/pm.pm_478_18  
Objective: To study the protective effect of Schisandra chinensis extracts (SCEs) on the RINm5F cells against H2O2-induced oxidative damage and to provide a basis for the study of the role of SCEs in the prevention and treatment of diabetic oxidative stress. Materials and Methods: We performed ultrasonic preparation of SCEs and verified the presence of 8 lignans by high-performance liquid chromatography (HPLC). An in vitro model of H2O2-induced oxidative damage was established using RINm5F cells and treated with various concentrations of SCEs (high dose, medium dose, and low dose). The antioxidative activity of SCEs was observed and its mechanisms were investigated by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay, and measuring malondialdehyde (MDA) and superoxide dismutase assay (SOD) according to the instructions of kits. Results: The total lignan content in SCEs was 18.86 mg/mL; the concentration of 8 kinds of lignans were schisandrin, 5.99 mg/mL; schisandrol B, 3.27 mg/mL; schisantherin A, 1.99 mg/mL; schisanhenol, 0.94 mg/mL; anwulignan, below the detection limit; deoxyschisandrin, 1.33 mg/mL; schisandrin B, 3.95 mg/mL; and schisandrin C, 1.39 mg/mL. SCEs promoted proliferation of islet cells and exhibited a protective effect against H2O2-induced oxidative damage in islet cells. The cell survival rates in the high-dose and medium-dose SCEs groups were greater than that in the model group by 63.4% and 45.6%, respectively; the protective effect of SCEs against oxidative damage was dose dependent. The intracellular MDA content was significantly decreased (P < 0.05), the degree of oxidative damage was reduced, and the activity of superoxide dismutase was significantly increased (P < 0.05) in the SCEs groups. Conclusion: SCEs exhibited a dose-dependent protective effect against H2O2-induced oxidative damage in RINm5F cells. The underlying mechanism was related to the antioxidant activity of the SCEs. Abbreviation used: Schisandra chinensis Extracts (SCEs).
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