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   2020| Oct-Dec  | Volume 16 | Issue 72  
    Online since February 16, 2021

 
 
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ORIGINAL ARTICLES
Anwulignan from the fruits of Schisandra chinensis and its cytotoxicity against human cancer cell lines

Oct-Dec 2020, 16(72):695-699
DOI:10.4103/pm.pm_208_20  
Background: The fruits of Schisandra chinensis have been used traditionally as the medicinal materials in East Asia. Their extracts have been reported to be cytotoxic to several cancer cells and phytochemical research has identified numerous cytotoxic lignans. Objectives: We reviewed for the effects of cytotoxic lignans against various human cancer cells. Materials and Methods: Solvent extraction of fruits in aqueous methanol and fractionation of the extract into H2O, n-BuOH, and EtOAc fractions (Fr) were carried out. Column chromatography (CC) of the non-polar EtOAc Fr, which indicated cytotoxicity to cancer cells, was performed to isolate the cytotoxic lignin. They were identified on the basis of the intensive spectroscopic interpretation of IR, 1D-, 2D-NMR, and mass spectrometry data. The cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and cell staining using Hoechst 32258 and propidium iodide. Results: Anwulignan was isolated and identified as a cytotoxic principal component. The IC50 value was calculated to be 22.01 ± 1.87, 156.04 ± 6.71, and 32.68 ± 2.21 μM for human stomach adenocarcinoma, human colon cancer (HT29), and human cervical cancer cells, respectively. Conclusion: Solvent extraction, systematic fractionation, and repeated CC for S. chinensis fruits led to the isolation of a lignin identified to be anwulignan based on various spectroscopic analyses. Anwulignan demonstrated very high cytotoxicity to most human cancer cell lines, and cell death was induced by apoptosis.
  2,455 322 -
Aqueous leaf extract of Clinacanthus nutans inhibits growth and induces apoptosis via the intrinsic and extrinsic pathways in MDA-MB-231 human breast cancer cells

Oct-Dec 2020, 16(72):689-694
DOI:10.4103/pm.pm_121_20  
Background: Clinacanthus nutans possesses several reported biological activities against different human cancer cells. However, reports on the growth-inhibitory effect of C. nutans leaf extract on the aggressive triple-negative breast cancer cells and the mechanisms of induced-cell death in these cells are limited. Objectives: The study aimed to assess the anticancer efficacy and associated mechanisms of the crude aqueous extract of C. nutans leaves (cCN) in MDA-MB-231 triple-negative human breast cancer cells. Materials and Methods: The metabolic viability of the MDA-MB-231 cells following respective treatments with cCN was measured using an adenosine triphosphate luminescent assay. The mode of cell death in MDA-MB-231 cells induced by cCN was examined using a luminescence- and fluorescence-based assay and the mechanisms involved were evaluated by comparative analysis of gene expression by reverse transcription–quantitative polymerase chain reaction. Results: Dose- and time-dependent growth inhibition of MDA-MB-231 cells by cCN was observed (IC50: 191.20 μg/mL). cCN also induced apoptotic cell death in the treated cells via the intrinsic and extrinsic apoptosis pathways by affecting the mRNA expression levels of Bad, Bax, Bcl-2, Bcl-xL, and FasL. Conclusion: These results suggest that C. nutans can be used as a potential agent in the treatment and prevention of breast cancer.
  2,215 529 -
Inhibitory effects of triterpenoid betulin on inflammatory mediators inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor-alpha, interleukin-6, and proliferating cell nuclear antigen in 1,2-dimethylhydrazine-induced rat colon carcinogenesis
Jinfeng Yu, Min Li, Dong Zhan, Chang Shi, Le Fang, Chunmei Ban, Weifeng Zheng, Vishnu Priya Veeraraghavan, Surapaneni Krishna Mohan, Xiaoling Tang
Oct-Dec 2020, 16(72):836-842
DOI:10.4103/pm.pm_516_19  
Background: Colon carcinogenesis is the third most commonly occurring malignant neoplasm and the second most common cause of cancer-related deaths globally. Betulin (BE) is a pentacyclic lupane-type triterpenoid naturally dispersed in many plants. It is also recognized as betuline, betulinol, or betulinic alcohol. Objectives: In this study, we explored the anticancer, antiproliferative, and anti-inflammatory effects of BE on 1,2-dimethylhydrazine (DMH)-treated rat model of colon cancer. Materials and Methods: Colon cancer was induced by a subcutaneous injection of DMH (20 mg/kg bwt) once a week for the initial 4 weeks of the experiment. We analyzed body weight, tumor incidence, tumor volume, total number of tumors, thiobarbituric acid reactive substances (TBARS), and levels of antioxidants (glutathione peroxidase, glutathione, catalase, and superoxide dismutase), bacterial enzymes (β-glucuronidase and mucinase), Phase I (cytochrome P450 and cytochrome b5) and Phase II (GST and Glutathione reductase (GR)) detoxification enzymes, and inflammatory (cyclooxygenase-2, interleukin-1 beta [IL-1β], inducible nitric oxide synthase, tumor necrosis factor-alpha, and IL-6) and cell proliferative (cyclin D1 and proliferating cell nuclear antigen) markers. We also assessed the histopathological alterations found in experimental and control rats. Results: We observed decreased body weight and levels of antioxidants and Phase II enzymes; augmented tumor incidence, tumor volume, total number of tumors, Phase I enzymes, TBARS, and levels of bacterial enzymes; and irregular histopathological changes in DMH-treated rats. Moreover, the Western blotting analysis of colon tissues revealed upregulation of inflammatory and cell proliferative markers in DMH-treated rats. Oral supplementation of 20 mg/kg bwt BE led to inhibition of tumor formation and inflammation, regulation of cell proliferation, and restoration of biochemical parameters. Our findings were supported by histopathological analysis. Conclusion: Our results suggested that BE exhibited anticancer, anti-inflammatory, and antiproliferative effects against DMH-induced colon cancer in rats.
  1,745 167 -
Ethanol extract of Thuja orientalis L. seeds ameliorated skin lesions in a dinitrofluorobenzene-induced mouse model of contact dermatitis

Oct-Dec 2020, 16(72):700-705
DOI:10.4103/pm.pm_245_20  
Background: Seed oils like sesame and camellia oils are used in herbal medicine to treat skin diseases and it has been reported that the seeds of Thuja orientalis have antioxidant, antibacterial, anti-inflammatory, and anti-allergic effects. Objectives: We investigate the anti-inflammatory effects of an ethanol extract of the seeds of T. orientalis (EETO) on a dinitrofluorobenzene-induced mouse model of contact dermatitis (CD). Materials and Methods: We investigated the effects of EETO on CD lesions and skin thickness, skin color, histopathologic findings, and cytokine production in mouse skins. Results: EETO ameliorated skin roughness, excoriation, scabs, and erythema. EETO also reduced erythema, melanin index, and inhibited skin thickness increases in CD mice. In addition, histopathologic examinations showed that EETO decreased epidermal hyperplasia, keratinization, and the productions of inflammatory cytokines (i.e., interleukin [IL]-6, tumor necrosis factor [TNF]-α, and monocyte chemotactic protein [MCP]-1). Conclusion: These results suggest that ethanolic extracts of the seeds of T. orientalis might provide relatively safe treatments for CD and that its therapeutic effect is exerted through inhibitions of IL-6, TNF-α, and MCP-1.
  1,624 271 1
Bioactivity-guided isolation, characterization, and estimation of esculetin – A Potential Marker from Launaea pinnatifida cass

Oct-Dec 2020, 16(72):713-721
DOI:10.4103/pm.pm_264_20  
Background: Loss of expertise in Sanskrit language and medicinal plant identification as per ayurvedic texts has resulted in the creation of “Controversial Drugs,” i.e., plants with different botanical identities being used by the same name Launaea pinnatifida Cass. It is one of the species of one such traditional Indian ayurvedic herb known as “Gojihva.” As per the ayurvedic and modern research, Gojihva possesses hepatoprotective, diuretic, galactagogue, and blood-purifying properties. The present study was aimed at bioactivity-guided isolation, characterization, and estimation of a marker compound obtained from L. pinnatifida. Materials and Methods: Fractions were obtained through the successive solvent extraction technique and subjected to in vitro antioxidant and hepatoprotective bioassays which signified that the methanol extract of leaves had significantly better pharmacological activity. Hence, the methanol extract was subjected to further isolation of the active compound. Estimation of the isolated compound was carried out by high-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography fingerprinting. Results: A marker compound was isolated from chloroform:ethyl acetate (90:10) fraction and elucidated to be esculetin based on its spectral analysis (Fourier transform infrared, liquid chromatography–mass spectroscopy, 13C nuclear magnetic resonance, and ultraviolet). The percentage purity and yield of esculetin in methanol extract of leaf fractions was found to be 99.75% and 0.927%, respectively, by HPLC. Conclusion: Esculetin has been reported for the first time in L. pinnatifida. Hence, it can be used as the bioactive marker for this plant. This marker can also be useful in the development of hepatoprotective formulations as well as standardization and quality control of L. pinnatifida formulations.
  1,334 234 2
Preclinical study on effects of Acalypha indica on streptozotocin-induced liver damage in diabetic rats

Oct-Dec 2020, 16(72):722-729
DOI:10.4103/pm.pm_272_20  
Background: Diabetes is one of the most common non-communicable global diseases. It is the fourth leading cause of death, thus posing a serious threat to human beings. The conventional drugs developed along the principles of Western medicine are often limited in efficacy in diabetes management. However, significant evidences suggest that polyphenol-rich diet has the ability to protect diabetes complications. Objective: In this study we investigated the quantifying and profiling of phenols in Acalypha indica polyphenol-rich fraction (PLF) and their therapeutic potential on hepatic tissue damage in streptozotocin (STZ) induced diabetic rats. Materials and Methods: In this study,PLF was subjected the gas chromatography–mass spectrometry and High Resolution Liquid Chromatograph Mass Spectrometer HR-LC-MS and evaluate the liver biomarkers includes aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), bilirubin content, lipid profiles like low-density lipoprotein (LDL), total cholesterol (TC), triglycerides (TGs), and high-density lipoprotein (HDL), insulin and body weights in diabetic, PLF, and glibenclamide-treated diabetic and normal rats. Results: The result showed that PLF was significantly decreased the blood glucose levels, liver biomarkers such as AST, ALT, ALP, bilirubin content, lipid profiles like LDL, TC, and TGs and decreased the tissue damage in diabetic rats. Subsequently, insulin, body weight, and HDL levels were increased in diabetic rats. Subsequently, insulin, body weight, and HDL levels were increased. Conclusion: We concluded that Ai (Acalypha indica ) leaves have a rich number of polyphenolic compounds and those were may be protected from STZ-induced toxicity in liver of diabetic rats. Further research is necessary for this area to isolate bioactive polyphenolic compounds from Ai leaves and to be validated against STZ-intoxicated liver damage with assessing mechanism of action.
  1,384 165 -
Simultaneous determination of two diterpenoids, continentalic acid and kaurenoic acid, in the water extract of Aralia continentalis and their wound-healing activity

Oct-Dec 2020, 16(72):745-749
DOI:10.4103/pm.pm_344_20  
Background: The roots of Aralia continentalis have been used in traditional Korean medicine to treat pain, lumbago, lameness, and rheumatism. Objective: Simultaneous determination of two diterpenoids; continentalic acid (CA) and kaurenoic acid, in the water extract of A. continentalis using the high-performance liquid chromatography (HPLC) was established for quality control, as well as evaluation of their wound-healing activity. Materials and Methods: The separation was conducted on YMC hydrosphere C18 column with isocratic elution of 65% acetonitrile. Furthermore, the isolated diterpenoids were screened for their wound-healing activity using keratinocytes (HaCaT cells). Results: The calibration curves were linear over the established range with R2 > 0.9999. The intraday and interday RSDs for each compound were 0.13%–0.89% and 0.14%–0.73%, respectively. The limits of detection and limits of quantification for the two tested diterpenoids were 0.0912, 0.0018 μg/μL, and 0.2764, 0.0056 μg/μL, respectively. In addition, the CA-treated groups showed accelerated wound closure compared to the control group. Conclusion: The HPLC method could be accomplished to the quality control and stable experiment for the preparations consisted of two diterpenoids. Compared to the control group, the CA-treated group showed that wound closure was accelerated, indicating its potential in promoting migration of skin cells which is the most important step of wound closure.
  1,320 228 1
Chemical composition and biological potential of a chloroform fraction from the leaves of marine plant Syringodium filiforme Kützing
Kethia Gonzalez Garcia, David Marrero Delange, Yasnay Hernández Rivera, Yulexi Acosta Suárez, Richard Gutiérrez Cuesta, Mario Riera-Romo, Olga Echemendia, Lívia Macedo Dutra, Jackson Roberto Guedes Da Silva Almeida, Dayana Pérez-martínez, Laurent Picot, Idania Rodeiro Guerra
Oct-Dec 2020, 16(72):750-756
DOI:10.4103/pm.pm_24_20  
Background: Seagrasses are an important component of Nearshore marine ecosystems and a rich source of secondary metabolites with important pharmacological properties. Materials and Methods: In this work, crude hydroethanolic extract and chloroform fraction (SfCHCl3) from marine plant Syringodium filiforme were evaluated for antimicrobial and cytotoxic potentials. In addition, the chemical composition of chloroform fraction was determined by gas chromatography-mass spectrometry (GC-MS) analysis. Results: GC-MS analysis allowed the identification of 68 compounds in the SfCHCl3, where palmitic acid (39.18%) was the main component. The evaluation of antibacterial activity of SfCHCl3 showed good activity against strains of Staphylococcus aureus (ATCC 6538), Pseudomonas aeruginosa (ATCC 9027), and Salmonella typhi (ATCC 9992v), and it was less active against Escherichia coli (ATCC 10576) and Candida albicans (ATCC 10231), while the crude extract showed low antibacterial activity. Furthermore, an important cytotoxic effect of SfCHCl3 in the A549 human lung carcinoma cell line was evidenced, which were similar to the cytotoxicity of cisplatin in the same cell line. Conclusion: The results suggest that compounds may play an important role in the antimicrobial and cytotoxic effects observed for this species. In addition, this research contributes to the chemotaxonomic characterization of S. filiforme and validates this species as a potential source of natural antimicrobial and cytotoxic molecules.
  1,310 195 1
Isolation of new diterpene from methanolic extract of Capsicum annuum Linn. fruits
Neeraj Kumar Fuloria, Shivkanya Fuloria, Vijay Kumar Sharma, Mohammed Ali, Amit Singh, Pradeep Kumar Sharma
Oct-Dec 2020, 16(72):730-732
DOI:10.4103/pm.pm_250_20  
Background: Capsicum annuum (CA) fruits are consumed in the human diet for more than six centuries. Less research evidence for critical isolation experiment over CA fruits methanolic extract (CAFME) is an important concern for the investigators. Objectives: The present investigation was intended to explore the CAFME for the presence of any new phytoisolate. Materials and Methods: The study involved the preparation of CAFME, followed by critical isolation through normal phase silica-gel based column chromatography using various combinations of petroleum ether and chloroform. The phytoisolate was characterized using ultraviolet, Fourier transformed infrared, Nuclear magnetic resonance (1H, 13C, Distortion less enhancement by polarization transfer and correlation spectroscopy), and mass spectrometry. Results: The chromatographic critical isolation and spectrometric experiment over CAFME offered a new phytoisolate characterized as 3,11,15-trimethyl-14 β-hydroxy-n-hexadeca-7-en-4,18-olide (CA-1). Conclusion: The present study concludes the isolation of a new diterpene 3,11,15-trimethyl-14 β-hydroxy-n-hexadeca-7-en-4,18-olide (CA-1) for the first time in CAFME. The present study also recommends that in future this new phytoisolate should be further standardized and explored for its therapeutic properties.
  1,288 190 2
Acute oral toxicity studies of methanolic extract and chloroform fraction of methanolic extract of seeds of Annona muricata
Bibu John Kariyil, PT A. Usha
Oct-Dec 2020, 16(72):757-763
DOI:10.4103/pm.pm_118_20  
Background: Natural products are considered to be safe and efficacious. Adequate proof of the safety of these herbal drugs is lacking. Objectives: The present study was undertaken to evaluate the acute oral toxicity of methanolic extract of seeds of Annona muricata (MAM) and chloroform fraction of MAM and to derive the lethal dose 50 (LD50) of chloroform fraction of methanol extract of seeds of A. muricata (CMAM). Materials and Methods: Acute toxicity studies of MAM and CMAM were conducted as per the Organisation for Economic Co-operation and Development (OECD) guidelines 420. LD50 of CMAM in rats and mice was estimated as per OECD guidelines 425. The toxic response was evaluated in vitro using hemolytic assay in rat and human erythrocytes. Results: The acute toxicity study done using OECD 420 guideline revealed that MAM and CMAM were safe at 300 mg/kg and produced mortality at 2000 and 5000 mg/kg. On gross examination, congestion of cranial, mesenteric, and uterine blood vessels was observed. As per OECD guidelines 425, CMAM was safe up to 980 mg/kg in rats and up to 175 mg/kg in mice and LD50 value was estimated to be 1310 mg/kg in rats and 310 mg/kg in mice. In vitro hemolytic assay showed that the plant fraction produced hemolysis. Conclusion: The present study concluded the toxicity produced by A. muricata seeds in the vascular system might be due to oxidative injury caused by the phytochemical constituents and it is essential to derive the safe pharmacological dose whenever the plant material is tested for efficacy studies.
  1,272 182 -
The effects of Elateriospermum tapos against obese maternal rat in mitigating obesity development among their adult female offspring

Oct-Dec 2020, 16(72):706-712
DOI:10.4103/pm.pm_142_20  
Background: Maternal obesity is a metabolic disorder described by chronic inflammation and dyslipidemia and influences non-communicable diseases in offspring. Elateriospermum tapos is a plant containing high alpha-linolenic acid with antiobesity effect. Objectives: This study was aim to investigate the effect of E. tapos supplementation in obese rat prior pregnancy on the female offspring's body weight, liver and kidney profile, and histological changes of heart, liver, kidney, and retroperitoneal white adipose tissue (RpWAT). Materials and Methods: Thirty female Sprague Dawley rats were used in this study. Six rats were assigned to dams normal diet group fed with standard chow diet. The remaining rats were fed with high-fat and cafeteria diet (HFCD) to generate obesity for 5 weeks. The obese rats were further divided into 4 groups; group1; Dams Negative Control (DNC, normal saline only), group 2; Dams Positive Control (DPC, Orlistat drug 200mg/kg), Dams Treatment 1 (DTX1, E. tapos seed 200 mg/kg) and Dams Treatment 2 (DTX2, E. tapos shell 200mg/kg) extracts for 6 weeks daily prior to mating. The weaning offspring were then designated into 6 groups according to their dam's group (n = 6/group). The offspring were culled at 12th week of age in order to have their blood sample and organs. Results: Adult offspring from DTX2 group showed a decrease in weight gain, alanine aminotransferase, and creatinine level compared to adult offspring from DNC. Histological examination showed that liver and RpWAT of offspring from DTX1 and DTX2 groups were comparable with normal diet group, indicating that the extracts were capable in reversing the high fat diet effect to the organ. Conclusion: E. tapos shell supplementation was effective in reducing the development of obesity with no harmful effect among adult female rat offspring.
  1,245 187 1
Isolation, structural characteristics, and in vitro and in vivo antioxidant activity of the acid polysaccharide isolated from Pholiota nameko

Oct-Dec 2020, 16(72):738-744
DOI:10.4103/pm.pm_109_20  
Background: Previous studies have revealed that edible fungi are rich in polysaccharides, which typically have strong biological activity. Pholiota nameko is one of the main cultivated edible fungi and an important economic crop. Objectives: In order to screen a polysaccharide with antioxidant activity from Pholiota nameko. Methods and Materials: A water-soluble acidic P. nameko polysaccharide (PNP) was found from extraction with alkaline water. After purification by chromatography, the structural features of PNP were analyzed by chromatographic, spectral, & nuclear magnetic resonance (NMR) spectroscopic analyses and the content of sulfate & uronic acid in PNP was determined concurrently. Results: The content of sulfate and uronic acid was 1.756 and 7.173 w/w, respectively. The molecular weight of PNP was 1.05 × 106 Da by gel permeation chromatography. The Fourier transform infrared (FTIR) and NMR spectra displayed that the following sugar chains might be present in PNP: β-1,4-D-Manp, →4)-β-D-Manp-(1→, β-1,3-D-Glcp, β-D-Xylp-(1→. Evaluation of in vitro antioxidant activity showed that PNP had a strong scavenging ability to scavenge ABTS+, hydroxyl, and superoxide anion radicals. The results of our in vivo experiments exposed that PNP might significantly lessen the formation of malondialdehyde. High levels of PNP can meaningfully increase the activity of catalase and superoxide dismutase activity in mice serum. In addition, PNP can significantly increase the activity of GSH-Px, effectively protect the structure and function of the cell membrane. Conclusion: PNP has antioxidant effect and can be explored as a potential antioxidant.
  1,162 193 2
Anti-inflammatory and anti-cell proliferative effects of dieckol in the prevention and treatment of colon cancer induced by 1,2-dimethyl hydrazine in experimental animals
Baoxiu Yang, Ying Li, Zhengbo Yang, Legang Xue, Minqin Zhang, Guoliang Chen, Arunachalam Chinnathambi, Tahani Awad Alahmadi, Xu Liu
Oct-Dec 2020, 16(72):851-858
DOI:10.4103/pm.pm_165_20  
Objectives: Colon carcinogenesis is a major cause of mortality and morbidity in developing and developed countries, and its etiology is familiar to be a grouping of environmental and nutritional factors, hereditary factors, and deficiency of physical activity. In the present study, we investigated the anti-cell proliferative and anti-inflammatory effects of dieckol (DEK) on 1,2-dimethylhydrazine (DMH)-treated colon carcinogenesis in investigational rats. Materials and Methods: Colon carcinogenesis was induced with DMH (20 mg/kg body weight) by subcutaneous injection once weekly. We analyzed body weight, tumor incidence, tumor volume, total number of tumors, thiobarbituric acid-reactive substances (TBARS), antioxidants (glutathione peroxidase, glutathione, catalase, and superoxide dismutase), Phase II (glutathione reductase and glutathione S-transferase) and Phase I (Cyt-b5 and CYP450) biotransformation enzymes, and histopathological alterations in the control and investigational rats. Moreover, the inflammatory (interleukin [IL] IL-1β, cyclooxygenase-2, tumor necrosis factor-alpha, IL-6, and inducible nitric oxide synthase) and cell proliferative (cyclin D1 and PCNA) markers were analyzed by Western blot technique in experimental and control rats. Results: We noted decreased body weight, antioxidants and Phase I and II enzymes, augmented tumor incidence, tumor volume, total number of tumors, TBARS, and irregular histopathological changes in DMH-induced animals. In addition, the Western blotting analysis of colon tissues showed an upregulation of inflammatory and cell proliferative markers in DMH-treated rats. Oral supplementation of DEK inhibited the tumor formation, controlled inflammation, cell proliferation, and restoration of biochemical parameters, and it was supported by the histopathological analysis. Conclusion: Findings from the study suggest that DEK demonstrated anticancer, anti-inflammatory, and anti-cell proliferative effects against DMH-treated colon carcinogenesis in rats.
  1,160 155 2
Terfezia claveryi and Terfezia boudieri extracts: An antimicrobial and molecular assay on clinical isolates associated with eye infections
Lorina Ineta Badger-Emeka, Promise Madu Emeka, Saif Aldossari, Hany Ezzat Khalil
Oct-Dec 2020, 16(72):780-788
DOI:10.4103/pm.pm_199_20  
Background: Ocular infections are capable of spreading to different anatomical sites of the eyes and, if not appropriately treated, can lead to blindness. The emergence of difficult to treat microbial infections has led to the search of alternatives from natural sources. Objectives: The antimicrobial effects of Terfezia claveryi and Terfezia boudieri (T. boudieri) against bacteria isolates associated with eye infections and their molecular mechanism were investigated. Materials and Methods: Crude aqueous and methanolic extracts, including fractions of chloroform, petroleum, and ethyl acetate of T. claveryi and T. boudieri, were used for the investigation. Bacterial isolation and identification were carried out using basic microbiological and biochemical techniques. scanning electron microscopy.(SEM) and molecular docking were used to adduce possible antimicrobial mechanism of these extracts and their fractions. Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus hominis, Staphylococcus lugdunensis, Serratia odorifera, Serratia liquefaciens, Pseudomonas stutzeri, Pseudomonas oryzihabitans, Proteus mirabililis, Kocuria kristinae, Kocuria rosea, and Micrococcus luteus were isolated from patients with ocular infections. Results: Isolates were resistant to benzylpenicillin (78.0%), rifampicin (57.0%), tetracycline (56.0%), clindamycin (33.3%), and tigecycline (24.0%). Furthermore, the percentage resistance to gentamicin and ciprofloxacin was 13.0% each. All isolates were susceptible to extracts/fractions of T. claveryi and T. boudieri. Docking analysis showed binding with surface protein Sortase A of Staphylococcus aureus, indicating that stigmasterol, the active compound in both Terfezia species, interacted with valine amino acid 110. SEM imaging showed morphological alterations in treated isolated Staphylococcal species. Conclusion: Therefore, extracts of both Terfezia species have demonstrated the potential to possess antibacterial activity, which can be further exploited for clinical use.
  1,072 187 1
Senna alexandriana mill as a potential inhibitor for quorum sensing-controlled virulence factors and biofilm formation in Pseudomonas aeruginosa PAO1
Manmohit Kalia, Devendra Singh, Deepmala Sharma, Shahid Suhail Narvi, Vishnu Agarwal
Oct-Dec 2020, 16(72):797-802
DOI:10.4103/pm.pm_315_20  
Background: Senna alexandriana Mill is a medicinally important plant used in laxatives and also possesses potent antimicrobial compounds, but its effect on the Pseudomonas aeruginosa was never analyzed. Aim: This study was carried out with the objective to attenuate quorum sensing (QS)-controlled virulence factors in P. aeruginosa and its biofilm. Methods: In the present study, acetone and hexane extracts of S. alexandriana mill were tested at sub-minimum inhibitory concentration concentrations for their anti-QS activity using bioreporter strain the Chromobacterium violaceum CV026. The effect of S. alexandriana Mill on different virulence phenotypes such as pyocyanin and protease production was investigated. Finally, the effect of the extract on P. aeruginosa, biofilm formation was also determined using the spectrophotometry-based method and microscopic analysis. Results: The extracts were tested for their activities against QS-controlled virulence factors of P. aeruginosa at 600 μg/ml and 800 μg/ml acetone and hexane extract concentration, respectively. Results showed that both the extracts have an inhibitory effect on different virulence factors of P. aeruginosa, including pyocyanin, protease production, and biofilm formation, without affecting the growth. The P. aeruginosa biofilm was found to be decreased by 75% and 62% in acetone and hexane extract presence. Conclusion: The results revealed that both acetone and hexane extracts showed anti-biofilm and anti-QS activity. It depicted the potential of S. alexandriana Mill against P. aeruginosa infection, its biofilm, and QS.
  1,076 157 3
Factorial design-guided optimization of extraction of therapeutically active furanocoumarin khellin from Ammi majus L. fruits
Hina Kausar, Lubna Abidin, Mohd Mujeeb, Mohd Aqil, Ozair Alam
Oct-Dec 2020, 16(72):830-835
DOI:10.4103/pm.pm_295_20  
Background: The use of Ammi majus L. for the treatment of various diseases has been reported by many. Its umbelliferous fruits possess khellin that is responsible for various pharmacological activities and used in treating psoriasis and vitiligo. Objectives: The study objective was to analyze the optimization of extraction parameters of khellin from A. majus L. by response surface methodology (RSM). Materials and Methods: Box–Behnken Design (BBD) of RSM was used for optimization needs. Quantification of khellin in extracts was done by high performance thin-layer chromatography using a mix of ethyl acetate: toluene: formic acid, and the peaks were monitored at 254 nm. Results: Among the explored traditional modes and ultrasound-assisted extraction (UAE), UAE was found to be the most ideal for khellin extraction, with methanol being the most suitable solvent. The applied BBD established a quadratic model for the experimental setup with the regression coefficient of 0.998. The optimal conditions were set up as – extraction temperature: 63.84°C, extraction time: 29.51 min. and solvent-to-drug ratio: 21.64 v/w, which yielded 6.21% w/w of khellin. Contrarily, under the modified experimental conditions, 6.86% w/w of khellin was extracted. Conclusion: It was concluded that all the variables studied significantly affected khellin yield. Moreover, a contemporary green extraction mode stood out to be the best for khellin extraction.
  1,086 123 2
Ameliorative effect of Tinospora tuberculata in insulin resistance: Potential roles of oxidative stress resistance and heat shock protein 70 (Hsp 70) modulation
PK Lakshmi, Shweta Kumar, Neha Dobriyal, Chandan Sahi, Rajesh Singh Pawar
Oct-Dec 2020, 16(72):764-773
DOI:10.4103/pm.pm_366_20  
Background: Tinospora tuberculata Bumee is a climbing vine distributed throughout the South Asian countries and it is known to be a hypoglycaemic agent. However, its role in cytoprotection and cellular oxidation is unclear. In the current investigation, cytoprotection against cellular stress and heat shock protein 70 (Hsp 70) modulation was investigated to understand the beneficial effect of butanolic fraction of T. tuberculata (TTB) in hyperglycemia and hyperlipidemia. Materials and Methods: The cytoprotective effect of TTB was studied on wild and knock-out yeast strain (Δtrx2). The effect of TTB on cell viability and the level of mitochondrial superoxide were determined to assess the antioxidant protection against reactive oxygen species assault. IR was induced in Wistar rats by high-fat diet and a low dose of streptozotocin. Hyperglycemic rats were orally treated with TTB (50 and 100 mg/kg body weight) or metformin (200 mg/kg b.w.) daily for 2 weeks. The effect of TTB on different parameters such as body weight, glucose levels, insulin resistance, glucose tolerance, lipid profile, antioxidant status, and hepatic Hsp 70 was analyzed in IR rats. TTB was characterized using high-resolution liquid chromatography-mass spectrometer (HR-LC/MS). Results: TTB demonstrated significant protection against the oxidative damage caused by H2O2 in both wild and Δtrx2 strains. In obese rats, TTB treatment reduced insulin resistance, improved glycaemic status, glucose tolerance, lipid profile, antioxidant level, and upregulated hepatic Hsp 70. HR-LC/MS analysis of TTB showed the presence of isoquinoline alkaloids (ambelline and reticuline), flavonoid glycoside (apiin), disaccharide carbohydrate (trehalose), cardenolide (calotropin), etc., Conclusion: The positive effect of the TTB on IR is attributed to its cytoprotective action against cellular stress and the activation of hepatic Hsp 70.
  1,030 101 -
Two new xeniolide diterpenes from the soft coral Xenia umbellata; displayed anti proliferative effects
Walied M Alarif, Mohamed A Ghandourah, Ahmed Abdel Lateff, Nahed O Bawakid, Ahmed Abdullah Alotaibi, Mohammad Y Alfaifi, Serag Eldin I. Elbehairi
Oct-Dec 2020, 16(72):774-779
DOI:10.4103/pm.pm_300_20  
Background and Objective: Xenia is an octocoral genus of family Xeniidae. It contains 98 species and is rich of xenican-type diterpenoids. These compounds play an important role in the biological activity of Xenia. Different bioactivities were reported, particularly, anticancer effects. Materials and Methods: A specimen of a Xenia umbellata was exhausted with organic solvents. Then, the organic extract has been fractionated and purified employing different chromatographic procedures. The spectral information obtained from different nuclear magnetic resonance experiments, mass spectroscopy, infrared, and ultraviolet was the key to elucidate the chemical structures. The anti-proliferative activities of all compounds have been evaluated against hepatocellular carcinoma (HepG2), prostate adenocarcinoma (PC-3), and colorectal adenocarcinoma (HT-29) cells. Results: Two new xeniolide-type diterpenes, xeniolide L (1) and xeniolide M (2), along with two known diterpenes, xeniolide K (3) and xeniumbellal (4) were isolated. Compounds 1–4 exhibited significant cytotoxic effect with IC50 values ranged from 0.17 ± 0.01 to 64.7 ± 0.40 μg/mL. Compound 1 displayed late apoptotic and necrotic effects in both HepG2 and PC-3, while 2 exhibited late apoptosis in HepG2 cells. Conclusion: The isolated xeniolide diterpenes displayed antiproliferative effects against tumor cells (HepG2, PC-3, and HT-29). The new compounds showed late apoptotic and necrotic effects in HepG2 cells.
  993 121 1
Limonin: A triterpenoid exerts protective effect during lipopolysaccharide stimulated inflammation in BV2 microglial cells
Shiyong Lu, Jixin Zhang, Xuemei Chen, Suresh Mickymaray, Zihao Liu
Oct-Dec 2020, 16(72):859-864
DOI:10.4103/pm.pm_304_19  
Objectives: Neuroinflammation mediated by the reactive oxygen and nitrogen species (ROS and RNS, respectively) is responsible for the production of cytokines and chemokines in the central nervous system, which causes glial activation and associated pathology. Limonin is an oxygenated triterpenoid dilactone obtained from the families Rutaceae and Meliaceae. It exerts anticancer, antioxidant, antiviral, antinociceptive, and anti-inflammatory effects. In this study, we intended to analyze the anti-inflammatory and neuroprotective activities of limonin against neuroinflammation induced by lipopolysaccharide (LPS) in an in vitro model with BV2 immortalized murine microglial cells. Materials and Methods: Cytotoxicity of limonin was investigated via 3-(4,5-dimethylthiazolyl-2)-2,5-dipheniltetrazolium bromide tetrazol assay, and anti-inflammatory activity was assessed by analyzing the status of ROS, nitric oxide (NO), prostaglandin E2 (PGE2), interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α using ELISA. The expression of inducible nitric oxide synthase (iNOS), TNF-α, IL-1β, and Cyclooxygenase (COX-2) was studied through immunoblot analysis. Results: Our results showed that pretreatment of LPS-induced microglial cells with limonin appreciably prevented the formation of ROS and prevented the accretion of NO, PGE2, IL-1β, IL-6, and TNF-α. Furthermore, limonin pretreatment significantly reduced the expressions of iNOS and COX-2. Conclusion: Our results indicate that limonin inhibits inflammatory response through repressing the formation of ROS, accretion and discharge of cytokines, and activation of inflammatory mediators. Limonin can be used as a neuro-protecting agent.
  976 125 2
Astaxanthin attenuates contrast-induced acute kidney injury through silent mating-type information regulation 2 homolog-1/peroxisome proliferator-activated receptor γ co-activator–α/NRF1 signaling pathway

Oct-Dec 2020, 16(72):733-737
DOI:10.4103/pm.pm_81_20  
Objectives: In this study, we established a model of contrast-induced acute kidney injury with iohexol. This model was used to investigate whether astaxanthin (AST) attenuates oxidative stress and apoptosis by activating silent mating-type information regulation 2 homolog-1 (SIRT1) signaling pathway in rat renal tubular epithelial cells (NRK-52E). Materials and Methods: NRK-52E cells were randomly divided into six groups: control group (CON group), vehicle (dimethyl sulfoxide group, iohexol group (I group), AST pretreatment group (AST + I group), AST plus nicotinamide (NA) co-pretreatment group (AST + NA + I group), and NA treatment group (NA + I group). The cellular activity was measured by cell counting kit-8. We estimated the levels of malonaldehyde (MDA) by the thiobarbituric acid method, and the level of intracellular reactive oxygen species (ROS) was measured by flow cytometry. Western blot analysis was conducted to detect the protein levels of SIRT1, peroxisome proliferator-activated receptor γ co-activator–α (PGC-1α), and NRF1. Results: Compared with the CON group, the I group showed suppressed cellular activity; increased levels of MDA and ROS; and decreased levels of SIRT1, PGC-1α, and NRF1 protein. Compared with the I group, AST + I pretreated group showed high cellular activity; low levels of MDA and ROS; and increased levels of SIRT1, PGC-1α, and NRF1 protein. SIRT1 inhibitor NA reversed the protective effect of AST. Compared with the AST + NA + I group, NA + I group showed low cellular activity; high levels of MDA and ROS; and low levels of SIRT1, PGC-1α, and NRF1 protein, which further confirmed the protective effect of AST. Conclusion: AST alleviated iohexol-induced NRK-52E cell injury by decreasing the formation of MDA and ROS and upregulating the SIRT1/PGC-1α/NRF1 signaling pathway.
  927 151 -
Antineoplastic potential of eupatilin against benzo[a]pyrene-induced lung carcinogenesis
Yanzhou Han, Zhiqing Zheng, Fanping Liu, Yanqing Tian, Lixin Bi, Sujuan Zhang
Oct-Dec 2020, 16(72):843-850
DOI:10.4103/pm.pm_162_20  
Background: Most of the conservative therapies used to treat lung cancer show serious side effects. In addition, the prevalence and death rates due to lung cancer have been increasing alarmingly across the globe. Eupatilin (EUP) is a naturally occurring flavone which is primarily the active ingredient of the traditional Chinese medicine Artemisia asiatica. Materials and Methods: In this study, we evaluated the antineoplastic effect of EUP against benzo(a)pyrene-induced lung cancer in Swiss albino mice. We analyzed the level of xenobiotics, liver dysfunction enzymes (LDEs), pro-inflammatory cytokines, and histology of the liver. Furthermore, we conducted in vitro experiments (A549 cells) to elucidate the amount of cell proliferation, apoptosis, and their markers (caspases 3 and 9). Results: The EUP (30 mg/kg bw) treatment of tumor-bearing mice with EUP revealed the normal levels of xenobiotic, LDEs, antioxidant enzymes, lipid peroxidation in the liver and further carcinoembryonic antigen, pro-inflammatory marker, and histology in lung tissues. EUP inhibited the proliferation of A549 cells and induced the formation of reactive oxygen species and apoptosis by upregulating the expression of caspases 3 and 9. Conclusion: Overall, these results substantiate the anti-neoplastic effects of EUP against carcinogen-induced lung cancer in in vitro and in vivo models.
  886 97 -
Two new flavonoid glycosides isolated from the fruits of Catalpa ovata
Liu-Qiang Zhang, Hai-Bing Xue, Wei-Liang Zhu, Yi-Ming Li, Kai-Xian Chen
Oct-Dec 2020, 16(72):812-816
DOI:10.4103/pm.pm_91_20  
Background: Catalpa ovata, widespread in China, Japan, and Korea, is receiving greater attention due to its potent pharmacological properties. Objectives: This communication report presents the results of chemical investigation and antioxidant capacity of the fruits of C. ovata. Materials and Methods: The polar fraction from the fruits of C. ovata was isolated using various chromatographic methods and their structures were identified through nuclear magnetic resonance spectrometry and high-resolution mass spectrometry, and the data were compared with the reported literature. Total antioxidant activity was analyzed using the total antioxidant capacity assay kit with a rapid ABTS method. Results: Cavonosides A and B (1 and 2), two new flavonoid glycosides, together with 13 known phenols (3–15), were obtained from the fruits of C. ovata. The antioxidant capacity of compounds 4, 7, 10, 13, and 15 was approximately equal to that of Vitamin C at the same concentration. Conclusion: This research work resulted in two new flavonoid glycosides (1 and 2) and 13 known phenols isolated from the fruits of C. ovata; among them, compounds 4, 7–10, and 13 were newly isolated from this genus. According to our results, the antioxidant capacity of compounds 4, 7, 10, 13 and 15 was approximately same as that of Vitamin C at the same concentration. We also discuss the structure–activity relationships of the isolated compounds.
  742 107 -
Synergistic and toxicity-attenuating effects of Periplaneta americana extract CII-3 combined with cisplatin on lewis lung cancer-bearing mice
Qiaoling Yang, Mili Shi, Guangming Liu, Fang Peng, Xiaobo Liu, Meixian Guo
Oct-Dec 2020, 16(72):817-829
DOI:10.4103/pm.pm_499_19  
Objective: The objective was to study the synergistic and attenuating effects of CII-3 combined with cisplatin. Materials and Methods: A Lewis tumor-bearing mouse model was established. After 15 days of continuous administration of CII-3 and cisplatin, the pathological changes in the tumor, liver, lung, and femur tissues were observed; the life prolongation rate, tumor inhibition rate, Q value, organ indices, spleen T- and B-lymphocyte proliferation activities, NK cell killing activity, the bone marrow cell proliferation rate and cell cycle phase, and the number of peripheral blood cells and bone marrow nucleated cells were measured. The expression of granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in mouse serum and bone marrow tissue was measured. Results: The combination of CII-3 and cisplatin could enhance the ability of cisplatin to inhibit tumor cell proliferation and protects the liver damage and femoral injury and could significantly increase the life prolongation rate; tumor inhibition rate of cisplatin; the liver, spleen, lung, and thymus indices; the T- and B-lymphocyte proliferation activity; the NK cell killing activity; and the number of peripheral blood cells and bone marrow nucleated cells. The combination of drugs can stimulate the transformation of bone marrow cells from S phase to G2/M phase, significantly increase the proliferation rate of bone marrow cells and the contents of G-CSF and GM-CSF in mouse serum, and downregulate the mRNA and protein levels of them in bone marrow tissue. Conclusion: These results suggest that CII-3 combined with cisplatin can significantly enhance the antitumor effect and reduce the toxicity and side effects of cisplatin in Lewis tumor-bearing mice.
  692 116 -
Juglanin attenuated adjuvant-induced arthritis via inactivating NF-κB/IκBα and a disintegrin-like and metalloproteinase domain with thrombospondin-1 repeat pathways in experimental rats
Chao Ning, Shaoqi Ning, Heng Luo, Lei Yan
Oct-Dec 2020, 16(72):803-811
DOI:10.4103/pm.pm_106_20  
Background: Rheumatoid arthritis (RA) is an autoimmune and inflammatory disease affecting multiple joints and worsening the quality of life. Juglanin, a natural aldose reductase inhibitor, has shown to exhibit anti-inflammatory, anti-nociceptive, and anti-oxidant properties. Objective: The present study was undertaken to evaluate their possible mechanism of action against adjuvant-induced arthritis (AIA), i.e., Freund's complete adjuvant (FCA)-induced arthritis, in experimental rats. Materials and Methods: FCA (0.1 mL) was administered into the subplantar region of female Wistar rats paw to induced AIA. The rats were treated with vehicle (distilled water, 10 mL/kg) or leflunomide (10 mg/kg), or juglanin (10, 20, and 40 mg/kg, respectively) orally for the next 16 days. Various biochemical, molecular, and histological parameters were evaluated to determine the potential of juglanin. Results: Subplantar administration of FCA resulted in a statistically significant (P < 0.05) induction of AIA reflected by alteration in paw volume, joint diameter, paw withdrawal threshold, and paw withdrawal latency, which was statistically significantly inhibited (P < 0.05) by juglanin (20 and 40 mg/kg). It also statistically significantly attenuated (P < 0.05) FCA-induced elevated hepatic oxido-nitrosative stress and mRNA expressions of tumor necrosis factor-alpha (TNF-α), interleukin (IL) IL-1β, IL-6, transforming growth factor-beta (TGF-β), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOs). Western blot analysis revealed that juglanin statistically significantly downregulated (P < 0.05) protein expressions of NF-κB, IκBα, a disintegrin-like and metalloproteinase domain with thrombospondin-1 repeats (ADAMTS)-4, and ADAMTS-5 in hepatic tissue. It also statistically significantly attenuated (P < 0.05) histopathological anomalies induced in the tibiotarsal joint. Conclusion: The present communication suggests that juglanin attenuated altered mechano-tactile allodynia and hyperalgesia via inhibition of elevated oxido-nitrosative stress, cytokines (TNF-α and ILs) levels, immune-inflammatory (TGF-β, NF-κB, Ikβα, COX-2, and iNOs) mediators, and ADAMTS, thus exerting its anti-arthritic potential.
  664 97 -
Anti-arthritic effect of neferine on adjuvant-induced experimental arthritis in rats via nuclear factor kappa B signaling pathway
Guoxi Gao, Chao Zhang
Oct-Dec 2020, 16(72):789-796
DOI:10.4103/pm.pm_538_19  
Background: Inflammation plays an important role in the progression of arthritis. The imbalance between pro-inflammatory and anti-inflammatory mediators regulates the induction and progression of arthritis. Nuclear factor kappa B (NF-κB) protein is ubiquitously present in all the cells. It controls the activation of immune system and regulates the inflammatory responses. Therefore, targeting NF-κB signaling pathway may be an effective strategy in treating arthritis. Neferine, a bisbenzylisoquinoline alkaloid, is present in the seeds of Nelumbo nucifera. Materials and Methods: In this study, arthritis was induced in rats with complete Freud's adjuvant (CFA, Group 2) and treated with neferine (Group 3) and diclofenac sodium (Group 4), respectively. The impact of neferine on arthritis was assessed by measuring the weight of organ, arthritis score index, hematological indices, and cytokines levels. Results: Hepatic enzymes were measured to assess the toxicity of neferine. The oxidative stress induced by CFA and the antioxidant property of neferine were estimated with biochemical assay, and their impact on the synovial tissue was confirmed with hematoxylin and eosin staining. To confirm the anti-inflammatory activity of neferine, the inflamed synovial tissue of normal and investigational animals was inspected through immunoblotting of NF-κB signaling proteins. Conclusion: Our overall results confirm that neferine effectively scavenged the oxidative stress induced by CFA and inhibited the NF-κB signaling, thereby alleviating the severity of arthritis. Histological analysis of the synovial tissue and arthritic score of arthritis-induced and neferine-treated rats authentically prove the potency of anti-arthritic drug in rat model.
  648 91 -
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