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   2017| October-December  | Volume 13 | Issue 52  
    Online since November 13, 2017

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Influence of flavonoids on mechanism of modulation of insulin secretion
Juliana Mikaelly Dias Soares, Ana Ediléia Barbosa Pereira Leal, Juliane Cabral Silva, Jackson R. G. S. Almeida, Helinando Pequeno de Oliveira
October-December 2017, 13(52):639-646
DOI:10.4103/pm.pm_87_17  PMID:29200726
Background: The development of alternatives for insulin secretion control in vivo or in vitro represents an important aspect to be investigated. In this direction, natural products have been progressively explored with this aim. In particular, flavonoids are potential candidates to act as insulin secretagogue. Objective: To study the influence of flavonoid on overall modulation mechanisms of insulin secretion. Methods: The research was conducted in the following databases and platforms: PubMed, Scopus, ISI Web of Knowledge, SciELO, LILACS, and ScienceDirect, and the MeSH terms used for the search were flavonoids, flavones, islets of Langerhans, and insulin-secreting cells. Results: Twelve articles were included and represent the basis of discussion on mechanisms of insulin secretion of flavonoids. Papers in ISI Web of Knowledge were in number of 1, Scopus 44, PubMed 264, ScienceDirect 511, and no papers from LILACS and SciELO databases. Conclusion: According to the literature, the majority of flavonoid subclasses can modulate insulin secretion through several pathways, in an indication that corresponding molecule is a potential candidate for active materials to be applied in the treatment of diabetes. Abbreviations used: KATP channels: ATP-sensitive K+ channels, GLUT4: Glucose transporter 4, ERK1/2: Extracellular signal-regulated protein kinases 1 and 2, L-VDCCs: L-type voltage-dependent Ca+2 channels, GLUT1: Glucose transporter 1, AMPK: Adenosine monophosphate-activated protein kinase, PTP1B: Protein tyrosine phosphatase 1B, GLUT2: Glucose transporter 2, cAMP: Cyclic adenosine monophosphate, PKA: Protein kinase A, PTK: Protein tyrosine kinase, CaMK II: Ca2+/calmodulin-dependent protein kinase II, GSIS: Glucose-stimulated insulin secretion, Insig-1: Insulin-induced gene 1, IRS-2: Insulin receptor substrate 2, PDX-1: Pancreatic and duodenal homeobox 1, SREBP-1c: Sterol regulatory element binding protein-1c, DMC: Dihydroxy-6'-methoxy-3',5'-dimethylchalcone, GLP-1: Glucagon-like peptide-1, GLP-1R: Glucagon-like peptide 1 receptor.
  22 5,132 382
Anti-cancer effects of polyphenolic compounds in epidermal growth factor receptor tyrosine kinase inhibitor-resistant non-small cell lung cancer
Hyungmin Jeong, Ai N. H. Phan, Jong-Whan Choi
October-December 2017, 13(52):595-599
DOI:10.4103/pm.pm_535_16  PMID:29200719
Background: Polyphenolic phytochemicals are natural compounds, easily found in fruits and vegetables. Importantly, polyphenols have been intensively studied as excellent antioxidant activity which contributes to anticancer function of the natural compounds. Lung cancer has been reported to mainly account for cancer-related deaths in the world. Moreover, epidermal growth factor receptor tyrosine kinase inhibitor (TKI) resistance is one of the biggest issues in cancer treatment, especially in nonsmall cell lung cancer (NSCLC). Even though several studies both in preclinical and clinical trials have showed promising therapeutic effects of polyphenolic compounds in anticancer therapy, the function of the natural compounds in TKI-resistant (TKIR) lung cancer remains poorly studied. Objective: The aim of this study is to screen polyphenolic compounds as potential anticancer adjuvants which suppress TKIR lung cancer. Materials and Methods: Colony formation and thiazolyl blue tetrazolium blue assay were performed in the pair-matched TKI-sensitive (TKIS) versus TKIR tumor cell lines to investigate the therapeutic effect of polyphenolic compounds in TKIR NSCLC. Results: Our data show that equol, kaempferol, resveratrol, and ellagic acid exhibit strong anticancer effect in HCC827 panel. Moreover, the inhibitory effect of most of tested polyphenolic compounds was highly selective for TKIR lung cancer cell line H1993 while sparing the TKIS one H2073. Conclusion: This study provides an important screening of potential polyphenolic compounds for drug development to overcome TKI resistance in advanced lung cancer. Abbreviations used: EGFR: Epidermal growth factor receptor, EMT: Epithelial-to-mesenchymal transition, GTP: Green tea polyphenols, IGF1R: Insulin-like growth factor 1 receptor, MET: Met proto-oncogene, MTT: Thiazolyl blue tetrazolium blue, NSCLC: Non-small cell lung cancer, ROS: Reactive oxygen species, RTK: Receptor tyrosine kinase, STAT3: Signal transducer and activator of transcription 3, TKIR: TKI-resistant, TKIs: Tyrosine kinase inhibitors, TKIS: TKI-sensitive.
  17 3,770 295
Amelioration of intestinal barrier dysfunction by berberine in the treatment of nonalcoholic fatty liver disease in rats
Donghao Li, Jimin Zheng, Yiting Hu, Hongtao Hou, Shurong Hao, Na Liu, Yuzhen Wang
October-December 2017, 13(52):677-682
DOI:10.4103/pm.pm_584_16  PMID:29200733
Objective: To investigate the effect of berberine (BBR) on intestinal barrier function in nonalcoholic fat liver disease (NAFLD) in rats. Materials and Methods: Rats were divided into three groups: normal diet group (control group [CON group]), high-fat diet feeding group (HFD group), and HFD with BBR group. After 8 weeks of HFD feeding, rats in the BBR group were given BBR intragastrically at a dose of 150 mg/kg daily for 4 weeks. The same volume of normal saline was given to the CON and HFD groups. Liver index was detected, and Sudan black B staining was used to study fatty degeneration, also the expression level of occluding and intestinal flora was analyzed. Results: BBR administration significantly reduced HFD-induced increase in body weight (CON group: 379.83 ± 61.51 g, HFD group: 485.24 ± 50.15 g, and BBR group: 428.60 ± 37.37 g). It obviously alleviated the HFD-induced liver fatty degeneration and histopathological changes of intestinal mucosa according to liver index low-density lipoprotein-cholesterol, high-density lipoprotein-cholesterol, and total cholesterol (P < 0.05). The triglyceride, alanine transaminase, and aspartate aminotransferase levels were greatly elevated after BBR treatment (P < 0.05); while endotoxin, intestinal fatty acid-binding protein, and tumor necrosis factor-α were significantly reduced (P < 0.05). Moreover, we found that BBR could obviously elevate the level of occludin and decrease the level of Faecalibacterium prausnitzii and upregulate the level of bacteroides. Conclusion: BBR provides significant protection in NAFLD through ameliorating intestinal barrier function. Abbreviations used: BBR: Berberine, NAFLD: Nonalcoholic fat liver disease, ALT: Alanine transaminase, AST: Aspartate aminotransferase, TG: Triglyceride, I-FABP: Intestinal-fatty acid-binding protein, IBD: Inflammatory bowel disease.
  15 3,835 339
Toxicity and detoxification effects of herbal Caowu via ultra performance liquid chromatography/mass spectrometry metabolomics analyzed using pattern recognition method
Yan Yan, Aihua Zhang, Hui Dong, Guangli Yan, Hui Sun, Xiuhong Wu, Ying Han, Xijun Wang
October-December 2017, 13(52):683-692
DOI:10.4103/pm.pm_475_16  PMID:29200734
Background: Caowu (Radix Aconiti kusnezoffii, CW), the root of Aconitum kusnezoffii Reichb., has widely used clinically in rheumatic arthritis, painful joints, and tumors for thousands of years. However, the toxicity of heart and central nervous system induced by CW still limited the application. Materials and Methods: Metabolomics was performed to identify the sensitive and reliable biomarkers and to characterize the phenotypically biochemical perturbations and potential mechanisms of CW-induced toxicity, and the detoxification by combinatorial intervention of CW with Gancao (Radix Glycyrrhizae) (CG), Baishao (Radix Paeoniae Alba) (CB), and Renshen (Radix Ginseng) (CR) was also analyzed by pattern recognition methods. Results: As a result, the metabolites were characterized and responsible for pentose and glucuronate interconversions, tryptophan metabolism, amino sugar and nucleotide sugar metabolism, taurine and hypotaurine metabolism, fructose and mannose metabolism, and starch and sucrose metabolism, six networks of which were the same to the metabolic pathways of Chuanwu (Radix Aconiti, CHW) group. The ascorbate and aldarate metabolism was also characterized by CW group. The urinary metabolomics also revealed CW-induced serious toxicity to heart and liver. Thirteen significant metabolites were identified and had validated as phenotypic toxicity biomarkers of CW, five biomarkers of which were commonly owned in Aconitum. The changes of toxicity metabolites obtained from combinatorial intervention of CG, CB, and CR also were analyzed to investigate the regulation degree of toxicity biomarkers adjusted by different combinatorial interventions at 6th month. Conclusion: Metabolomics analyses coupled with pattern recognition methods in the evaluation of drug toxicity and finding detoxification methods were highlighted in this work. Pattern recognition plot reflects the toxicity effects tendency of the urine metabolic fluctuations according to time after treatment of herbal Caowu. Abbreviations used: CW: Caowu (Radix Aconiti kusnezoffii); CHW: Chuanwu (Radix Aconiti); TCM: Traditional Chinese Medicine; CG: Caowu and Gancao; CB: Caowu and Baishao; CR: Caowu and Renshen; QC: Quality control; UPLC: Ultra performance liquid chromatography; MS: Mass spectrometry; PCA: Principal component analysis; PLS-DA: Partial least squares-discriminant analysis; OPLS: Orthogonal projection to latent structures analysis.
  12 3,586 267
Antioxidant and inhibitory effects of saponin extracts from Dianthus basuticus Burtt Davy on key enzymes implicated in type 2 diabetes In vitro
Mikhail Olugbemiro Nafiu, Anofi Omotayo Tom Ashafa
October-December 2017, 13(52):576-582
DOI:10.4103/pm.pm_583_16  PMID:29200716
Context: Dianthus basuticus is a plant of South African origin with various acclaimed pharmaceutical potentials. Aims: This study explored the antioxidant and antidiabetic activities of saponin extract from D. basuticus in vitro. Materials and Methods: Antioxidant activity of saponin was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and nitric oxide (*NO)-free radical scavenging activity while antidiabetic potentials were measured by the α-amylase and α-glucosidase inhibitory activities of the saponin extract. Results: The results showed that the saponin extract, compared with quercetin, displayed better DPPH (IC50 = 6.95 mg/ml) and NO (IC50 = 3.31 mg/ml) radical scavenging capabilities. Similarly, the saponin extracts elicited stronger α-glucosidase (IC50 = 3.80 mg/ml) and moderate α-amylase (IC50 = 4.18 mg/ml) inhibitory activities as compared to acarbose. Saponin exhibited a competitive mode of inhibition on α-amylase with same maximum velocity (Vmax) of 0.0093 mM/min for saponin compared with control 0.0095 mM/min and different the Michaelis constant (Km) values of 2.6 × 10-6 mM and 2.1 × 10-5 mM, respectively, while for α-glucosidase, the inhibition was uncompetitive, Vmax of 0.027 mM/min compared with control 0.039 mM/min and Km values of 1.02 × 10-6 mM and 1.38 × 10-6 mM, respectively. The gas chromatography-mass spectrometric analysis revealed the presence of bioactive like β- and α-amyrin, 3-O-methyl-D-glucose, methyl commate, and olean-12-en-3-beta-ol. Conclusion: Overall, the data suggested that the saponin extract from D. basuticus has potentials as natural antioxidants and antidiabetics. Abbreviations used: DPPH: 2,2-diphenyl-1-picrylhydrazyl, Km: The Michaelis constant, Vmax: Maximum velocity, ROS: Reactive oxygen species, NIDDM: Non-insulin-dependent diabetes mellitus, UFS: University of the Free State, GC-MS: Gas chromatography-mass spectrometric, MS: Mass spectrometry, NIST: National Institute of Standards and Technology, DNS: 3,5-dinitrosalicylic acid, NO: Nitric oxide, RNS: Reactive nitrogen species, PNPG: p-Nitrophenyl-α-D-glucopyranoside.
  12 4,065 360
Hispidulin-7-O-neohesperidoside from Cirsium japonicum var. ussuriense attenuates the production of inflammatory mediators in LPS-induced raw 264.7 cells and HT-29 cells
Jong Cheol Park, Hyunji Yoo, Cho Een Kim, Sun-Yup Shim, Mina Lee
October-December 2017, 13(52):707-711
DOI:10.4103/0973-1296.218116  PMID:29200737
Background: Inflammatory bowel disease (IBD) is characterized by chronic inflammation of the gastrointestinal tract and involves secretion of inflammatory mediators. The flavone diglycoside hispidulin-7-O-neohesperidoside (HN) isolated from the methanolic extract of aerial parts of Cirsium japonicum var. ussuriense, but its pharmacologic activities, with the exception of alleviation of alcohol toxicity, have not been investigated to date. Objective: The aim of the present study was to investigate the anti-inflammatory activities of HN for the treatment of chronic inflammatory illnesses, including IBD. Materials and Methods: In lipopolysaccharide (LPS)-induced RAW264.7 cells and HT-29 cells, the effects of HN on cell viability and nitric oxide (NO) production were examined via MTT assay and the Griess reaction, respectively. The expression levels of interleukin (IL)-1α, IL-8, and tumor necrosis factor (TNF)-α and inducible nitric oxide synthase (iNOS) protein levels were measured by enzyme-linked immunosorbent assay and Western blotting, respectively. Results: HN concentration-dependently inhibited NO production in LPS-induced RAW 264.7 cells. Treatment with HN considerably downregulated the levels of the pro-inflammatory cytokines, IL-1β and TNF-α and the iNOS protein level in LPS-induced RAW 264.7 cells. Furthermore, HN inhibited the production of the chemotactic cytokine, IL-8, in LPS-induced HT-29 cells. Conclusion: HN has potential as an anti-inflammatory agent to prevent and/or treat IBD. Abbreviations used: IBD: Inflammatory bowel disease, HN: hispidulin-7-O-neohesperidoside, LPS: lipopolysaccharide, NO: nitric oxide, IL: interleukin, TNF: tumor necrosis factor, iNOS: inducible nitric oxide synthase, CD: Crohn's disease, UC: ulcerative colitis, RT: room temperature, DMEM: Dulbecco's modified Eagle's medium, FBS: fetal bovine serum, PBS: phosphate buffered saline, SDS: sodium dodecyl sulfate, PVDF: polyvinylidene difluoride, SD: standard deviation
  11 3,603 309
Superoxide scavenging and antiglycation activity of rhinacanthins-rich extract obtained from the leaves of Rhinacanthus nasutus
Muhammad Ajmal Shah, Haji Muhammad, Yasir Mehmood, Ruqaiya Khalil, Zaheer Ul-Haq, Pharkphoom Panichayupakaranant
October-December 2017, 13(52):652-658
DOI:10.4103/pm.pm_196_17  PMID:29200728
Background: Oxidative stress and nonenzymatic protein glycation lead to serious diabetic complications that increase the risk of mortality. Rhinacanthus nasutus leaf crude extracts are previously reported for their antidiabetic, antiglycation, and antioxidant potential. Objective: The present study was performed to prepare a standardized rhinacanthins-rich extract (RRE) and evaluate its superoxide scavenging and antiglycation effects as compared to its marker compounds, namely, rhinacanthin-C (RC), rhinacanthin-D (RD), and rhinacanthin-N (RN). Materials and Methods: RRE was obtained by microwave-assisted green extraction along with a simple step of fractionation using Amberlite® column. RC, RD, and RN were isolated from the RRE using silica gel column chromatography. Superoxide scavenging activity was performed by cyclic voltammetry, and fructose-mediated human serum albumin glycation model was used for antiglycation activity. In silico studies were conducted to identify the structure-activity relationships of rhinacanthins. Results: On the basis of kinetic measurements, RRE exhibited the most potent antioxidant activity via ErCi mechanism, with a 50% inhibitory concentration (IC50) value of 8.0 μg/mL, antioxidant capacity of 39439 M−1, and binding constant of 45709 M−1. Antiglycation assay showed that RRE exhibited almost equivalent glycation inhibitory effect to that of RC, with IC50values of 39.7 and 37.3 μg/mL, respectively, but higher than that of RD (IC50 of 50.4 μg/mL), RN (IC50 of 89.5 μg/mL), as well as the positive control, rutin (IC50of 41.5 μg/mL). Conclusions: The potent superoxide scavenging and albumin glycation inhibitory effect of RRE rationalized its therapeutic application in various chronic diseases, especially in the complications of diabetes. Abbreviations used: RRE: Rhinacanthins-rich extract; RC: Rhinacanthin-C; RD: Rhinacanthin-D; RN: Rhinacanthin-N; IC50: 50% inhibitory concentration; Kao: Antioxidant activity coefficient; Kb: Binding constant; ErCi: Reversible electron transfer followed by an irreversible chemical reaction; DM: Diabetes mellitus; AGEPs: Advanced glycation end products; NMR: Nuclear magnetic resonance; HPLC: High-performance liquid chromatography; CV: Cyclic voltammetry; DMSO: Dimethyl sulfoxide; Ipa: Anodic peak current; Ipc: Cathodic peak current; HSA: Human serum albumin; MOE: Molecular operating environment; PASSonline: Online prediction of activity spectra for substances.
  9 3,100 241
Extraction of chelerythrine and its effects on pathogenic fungus spore germination
Qinghui Wei, Min Zhao, Xiaoyan Li
October-December 2017, 13(52):600-606
DOI:10.4103/pm.pm_545_16  PMID:29200720
Background: Chemical fungicides are widely used to control crop diseases, but these chemicals have adverse effects. They destroy the ecological environment and even have toxic effects on human beings. In this context, the development of botanical pesticides is relevant. One potential botanical pesticide is chelerythrine, a main alkaloid of Chelidonium majus L., which has high antitumor, fungistasis, and antiphlogosis bioactivity. Objective: This study was designed to present an ultrasonic extraction method for chelerythrine and spore germination experiments to inhibit pathogenic fungi. Fungistasis of chelerythrine is now centralized in basic microbiology experiments, such as observing bacteriostatic rings. This study investigates chelerythrine based on pathogenic fungal spore germination and the influence of germ tube elongation. Materials and Methods: Samples of C. majus L., which were wild used in this experiment, were picked from Harbin experimental forest farm of Northeast Forestry University. An L9 (34) orthogonal experiment was designed to optimize the ultrasonic extraction method. All the plant pathogenic fungus strains used in the experiment were preservation strains of Northeast Forestry University Microbial preservation center. Pathogenic fungi were cultivated by joining chelerythrine with and observed germ tube growth and spore germination. Results: The optimum ultrasonication extraction process for chelerythrine has a liquid/solid ratio of 1:8, 35 min of extraction time, 85% of ultrasonic frequency, and 75% of ethanol concentration. When the concentration of chelerythrine was 1.7 × 10−2 mg/ml, the inhibition rates of Septoria microspora Speg. spores and Curvularia lunata were 96.67% and 84.94%, respectively. Moreover, when the concentration of chelerythrine was 1.7 × 10−6 mg/ml, the inhibition rates of S. microspora spores and C. lunata were 47.64% and 12.05%, respectively. Conclusion: Fungistasis activity reached a high level with 1.7 × 10−6 mg/ml of chelerythrine. Chelerythrine has the characteristics of less dosage and obvious fungistasis and has a good prospect for botanical fungicide development. Abbreviations used: C. majus L.: Chelidonium majus L.; Sphaerulina juglandis: S. juglandis; Septoria microspora Speg.: S. microspora; Fusarium oxysporum f. sp. Lycopersici: F. oxysporum f. sp. lycopersici; F. oxysporum f. cucumerinum: F. oxysporum f. cucumerinum; Curvularia lunata: C. lunata.
  8 2,697 200
Evaluation of hypolipidemic and antioxidant effects in phenol-rich fraction of Crataegus pinnatifida fruit in hyperlipidemia rats and identification of chemical composition by ultra-performance liquid chromatography coupled with quadropole time-of-flight mass spectrometry
Feng Shao, Lifei Gu, Huijuan Chen, Ronghua Liu, Huilian Huang, Lanying Chen, Ming Yang
October-December 2017, 13(52):725-731
DOI:10.4103/pm.pm_402_16  PMID:29200740
Background: Hawthorn (Crataegus pinnatifida) fruit has enjoyed a great popularity as a pleasant-tasting food associated with hypolipidemic and antioxidant effects. Objective: Our aim was to screen the effective fraction of hawthorn fruit in the treatment of hyperlipidemia rats. Materials and Methods: In this study, ethanol extract of hawthorn fruit (Fr.1) and four fractionated extracts (Fr.2, Fr.3, Fr.4, and Fr.5) were compared to total phenol content evaluated using Folin–Ciocalteu method, and hypolipidemic and antioxidant effects were assessed in hyperlipidemic rats. Results: Total phenol content of Fr.4 was higher than other fractions by at least 2 fold. Furthermore, this fraction possessed the strongest hypolipidemic and antioxidant effects in hyperlipidemic rats. On this basis, 15 phenolic compounds and four organic acids in Fr.4 were positively or tentatively identified using ultra-performance liquid chromatography coupled with quadropole time-of-flight mass spectrometry. In addition, 5-O-caffeoyl quinic acid butyl ester was first reported in hawthorn fruit. Conclusion: Phenol-rich fraction in hawthorn fruit exhibited satisfactory hypolipidemic and antioxidant effects, and this could be exploited for further promotion of functional foods. Abbreviations used: UPLC-Q-TOF-MS/MS: Ultra performance liquid chromatography coupled with quadropole time-of-flight mass spectrometry; TC: Total cholesterol; TG: Triglyceride; LDL-C: Low-density lipoprotein-cholesterol; HDL-C: High-density lipoprotein-cholesterol; GSH-Px: Glutathione peroxidase; SOD: Superoxide dismutase; MDA: Malondialdehyde; CAT: Catalase; NO: Nitric oxide; NOS: Nitric oxide synthase; ROS: Reactive oxygen species; •OOH: Superoxide anions, •OH: Hydroxyl radicals.
  8 3,029 242
Rapid quantitative analysis of naringenin in the fruit bodies of Inonotus vaninii by two-phase acid hydrolysis followed by reversed phase-high performance liquid chromatography-ultra violet
Xia Guohua, Ruirong Pan, Rui Bao, Yanru Ge, Cunshan Zhou, Yuping Shen
October-December 2017, 13(52):659-662
DOI:10.4103/pm.pm_350_16  PMID:29200729
Introduction: Sanghuang is one of mystical traditional Chinese medicines recorded earliest 2000 years ago, that included various fungi of Inonotus genus and was well-known for antitumor effect in modern medicine. Inonotus vaninii is grown in natural forest of Northeastern China merely and used as Sanghuang commercially, but it has no quality control specification until now. This study was to establish a rapid method of two-phase acid hydrolysis followed by reversed phase-high performance liquid chromatography-ultra violet (RP-HPLC-UV) to quantify naringenin in the fruit body of I. vaninii. Materials and Methods: Sample solution was prepared by pretreatment of raw material in two-phase acid hydrolysis and the hydrolysis technology was optimized. After reconstitution, analysis was performed using RP-HPLC-UV. The method validation was investigated and the naringenin content of sample and comparison were determined. Results: The naringenin was obtained by two-phase acid hydrolysis method, namely, 10.0 g of raw material was hydrolyzed in 200 mL of 1% sulfuric acid aqueous solution (v/v) and 400 mL of chloroform in oil bath at 110°C for 2 h. Good linearity (r = 0.9992) was achieved between concentration of analyte and peak area. The relative standard deviation (RSD) of precision was 2.47% and the RSD of naringenin contents for repeatability was 3.13%. The accuracy was supported with recoveries at 96.37%, 97.30%, and 99.31%. The sample solution prepared using the proposed method contained higher content of naringenin than conventional method and was stable for 8 h. Conclusion: Due to the high efficiency of sample preparation and high reliability of the HPLC method, it is feasible to use this method for routine analysis of naringenin in the fungus. Abbreviations used: RP-HPLC-UV: Reversed Phase-High Performance Liquid Chromatography-Ultra Violet, RSD: Relative Standard Deviation, EtOAc: Ethyl acetate, ACN: Acetonitrile, MeOH: Methanol, RH: Relative Humility.
  5 2,779 167
Therapeutic effects of Cyathula officinalis Kuan and its active fraction on acute blood stasis rat model and identification constituents by HPLC-QTOF/MS/MS
Yanmei Cao, Cuicui Gu, Fangli Zhao, Yuanlin Tang, Xiaobing Cui, Le Shi, Li Xu, Lian Yin
October-December 2017, 13(52):693-701
DOI:10.4103/pm.pm_560_16  PMID:29200735
Background: Cyathula officinalis Kuan is widely used in the clinics for the treatment of blood stasis in China. Objective: To evaluate the improving blood rheology and anti-inflammatory properties of C. officinalis Kuan extract (CO) and its active fraction (ACO) on acute blood stasis model Wistar rats and characterize the correlative constituents. Materials and Methods: CO at 0.26, 0.53, and 1.04 g/kg and ACO at 0.38, 0.75, and 1.5 g/kg were administered to acute blood stasis model Wistar rats for 3 days. Whole blood viscosity, plasma viscosity, and the levels of interleukin-6 (IL-6), nitric oxide (NO), tumor necrosis factor alpha (TNF-α), and cyclooxygenase-2 (COX-2) in the plasma were measured. HPLC-QTOF/MS/MS method was used to identify the major constituents of ACO; the properties of two representative components (cyasterone and chikusetsusaponin IV) from ACO on thrombin-induced human umbilical vein endothelial cells damage model were also assessed by the levels of thromboxane A2 (TXA2), endothelin (ET), malondialdehyde (MDA), COX-2, endothelial nitric oxide synthase (eNOS), and superoxide dismutase (SOD). Results: CO and ACO significantly reduced whole blood viscosity, plasma viscosity, and levels of IL-6, NO, TNF-α, and COX-2 in vivo. Forty compounds were identified from ACO, mainly as phytoecdysteroids and saponins. Cyasterone and chikusetsusaponin IV could significantly inhibit levels of TXA2, ET, MDA, and COX-2 and promote the activities of eNOS and SOD in vitro. Conclusion: CO and ACO possessed significant improving blood rheology and anti-inflammatory effects on acute blood stasis model rats and the representative components Cyasterone and chikusetsusaponin IV showed significant anti-inflammatory, antioxidant, and anticoagulant effects in vitro. Abbreviations used: TCM: Traditional Chinese Medicine, CO: Cyathula officinalis Kuan extract, ACO: Active fraction of Cyathula officinalis Kuan, ROS: Reactive oxygen species, IL-6: Interleukin-6, TNF-α: Tumor necrosis factor alpha, NO: Nitric oxide, COX-2: Cyclooxygenase-2, TXA2: Thromboxane A2, ET: Endothelin, MDA: Malondialdehyde, eNOS: Endothelial nitric oxide synthase, SOD: Superoxide dismutase, ESI: Electronic spray ionization, ELISA: Enzyme-linked immunosorbent assay, HUVECs: Human umbilical vein endothelial cells, DMEM: Dulbecco's modified Eagle medium, MMP: Matrix metalloproteinase.
  5 3,293 304
Comparative proteomic analysis of three gelatinous chinese medicines and their authentications by tryptic-digested peptides profiling using matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry
Huan Yang, Jie Zheng, Hai-yan Wang, Nan Li, Ya-ya Yang, Yu-ping Shen
October-December 2017, 13(52):663-667
DOI:10.4103/pm.pm_54_17  PMID:29200730
Background: Gelatinous Chinese medicines (GCMs) including Asini Corii Colla, Testudinis Carapacis ET Plastri Colla, and Cervi Cornus Colla, were made from reptile shell or mammalian skin or deer horn, and consumed as a popular tonic, as well as hemopoietic and hemostatic agents. Misuse of them would not exert their functions, and fake or adulterate products have caused drug market disorder and affected food and drug safety. GCMs are rich in denatured proteins, but insufficient in available DNA fragments, hence commonly used cytochrome c oxidase I barcoding was not successful for their authentication. Objective: In this study, we performed comparative proteomic analysis of them and their animal origins to identify the composition of intrinsic proteins for the first time. Materials and Methods: A reliable and convenient approach was proposed for their authentication, by the incorporation of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, two-dimensional electrophoresis, and matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry (MALDI-TOF/TOF-MS). Results: A total of 26 proteins were identified from medicinal parts of original animals, and GCMs proteins presented in a dispersive manner in electrophoresis analyses due to complicated changes in the structure of original proteins caused by long-term decoction and the addition of ingredients during their manufacturing. In addition, by comparison of MALDI-TOF/TOF-MS profiling, 19 signature peptide fragments originated from the protein of GCM products were selected according to criteria. Conclusion: These could assist in the discrimination and identification of adulterates of GCMs and other ACMs for their form of raw medicinal material, the pulverized, and even the complex. Abbreviations used: GCMs: Gelatinous Chinese medicines, COI: Cytochrome c oxidase I, SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis, 2-DE: Two-dimensional electrophoresis, MALDI-TOF/TOF-MS: Matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry, LC: Liquid chromatography, ChP: Chinese Pharmacopoeia, HPLC: High performance liquid chromatography, LC-ESI+-MS: Liquid chromatography-electro spray ionization-mass spectrometry, IEF: isoelectric focusing, HCCA: α-Cyano-4-hydroxycinnamic acid.
  4 2,554 192
In vitro and In vivo postprandial glycemic activity of Citrus limetta peel flour
José Miguel Flores-Fernandez, Carla Patricia Barragán-Álvarez, Nestor Emmanuel Díaz-Martínez, Socorro Villanueva-Rodríguez, Eduardo Padilla-Camberos
October-December 2017, 13(52):613-616
DOI:10.4103/pm.pm_158_17  PMID:29200722
Background: Previous studies of Citrus spp. peel have shown hypoglycemic and antioxidant activities. Citrus limetta has been studied for its therapeutic properties. Diabetes mellitus (DM) is a health problem in Mexico and worldwide, that takes a vital importance due to its high incidence. Recently, scientists have searched natural sources to control the disease. Materials and Methods: In this study, we evaluated the in vitro hypoglycemic activity and in vivo postprandial glycemic effect of C. limetta peel flour by glucose adsorption and retardation assays as well as postprandial serum glucose levels using a group of female Balb-c mice, respectively. Results: C. limetta peel flour showed a glucose adsorption capacity of 16.58 mM, having a similar effect regarding the positive control. The glucose diffusion in the dialysate was elevated, with a glucose dialysis retardation index of 33.79% in a period of 3 h, showing similar results to positive control. Postprandial serum glucose levels in the animal group treated with C. limetta peel flour showed a glucose level of 41.4 mg/dL, being this value significantly lower than negative control group and similar to positive control. Toxicity tests showed good tolerance to the dose of 2000 mg/kg. Conclusion: C. limetta peel flour could act as a source of functional compounds for the control of DM. Abbreviations used: CIATEJ: Center for Research and Assistance in Technology and Design of Jalisco; DM: Diabetes mellitus; FGC: Final glucose concentration; GDRI: Glucose dialysis retardation index; IGC: Initial glucose concentration; OECD: Organization for Economic Cooperation and Development.
  4 2,854 142
Mice behavioral phenotype changes after administration of Anani (Symphonia globulifera, clusiaceae), an alternative Latin American and African medicine
Ivana Barbosa Suffredini, Mateus Luís Barradas Paciencia, Ingrit E. C. Díaz, Sergio Alexandre Frana, Maria Martha Bernardi
October-December 2017, 13(52):617-626
DOI:10.4103/pm.pm_168_17  PMID:29200723
Background: Anani, (Symphonia globulifera, Clusiaceae), known as chewstick, is a traditional plant occurring in Africa and in Central and South Americas that is used against parasites and microorganisms. Although its use is popular in some of these countries, there is a lack of information related to its influence over behavioral phenotype (BP). Objective: The objective of this study is to evaluate the influence of the administration of the extract obtained from the aerial organs of Anani (EB1257) to male Balb-c mice over BP. Materials and Methods: Open cage observation, open field, and elevated-plus maze apparatuses were used. Evaluations were done 15, 30, 60, 120, and 180 min after intraperitoneal administration of Anani extract. Results: Impairment of general behavior activity, response to touch, tail squeeze, defecation, locomotion and rearing frequency were observed although no signs of hemorrhage or macroscopical alterations of internal organs. Anani is harmful, but not toxic if used in the appropriate doses, yet to be determined to male mice. Impairment of locomotion and defecation was observed, indicating some degree of influence over locomotion, but no alterations in anxiety levels were assessed. Three compounds were previously found in the plant-lupeol (1), β-amyrin (2) and 3-β-hydroxyglutin-5-ene (3), and one is being described for the first time to occur in the species: oleanolic acid (4). Conclusions: The present work contributes in the support of the rational use of Anani, an important Latin American and African alternative medicine, presenting findings that are being reported for the first time. Abbreviations used: BP: Behavioral phenotype; OF: Open field, EPM: Elevated-plus maze, MMA/ICMBio/SISBIO: Ministério do Meio Ambiente/Instituto Chico Mendes de Conservação da Biodiversidade/Sistema de Autorização e Informação em Biodiversidade, IBAMA/MMA/CGen: Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis/Ministério do Meio Ambiente/Conselho de Gestão do Patrimônio Genético, AM: Amazonas State, UNIP: Universidade Paulista, mg: milligram, kg: kilogram, I.P: Intraperitoneal, CEUA/ICS/UNIP: Comissão de Ética no Uso de Animais/Instituto de Ciências da Saúde/Universidade Paulista, LD: Lethal dose, NLD: Nonlethal dose, GBA: General behavior activity, FCHCL3: Fraction chloroform, FBuOH: Fraction buthanol, FH2O: Fraction water, FrHEX: Fraction hexane, FrDCM: Fraction dichloromethane, FrMeOH: Fraction methanol, 13C NMR: Carbon nuclear magnetic resonance, EPA: United States Environmental Protection Agency.
  3 2,370 134
Bioassay-guided isolation of neuroprotective fatty acids from Nigella sativa against 1-methyl-4-phenylpyridinium-induced neurotoxicity
Leila Hosseinzadeh, Hoda Monaghash, Farahnaz Ahmadi, Nastaran Ghiasvand, Yalda Shokoohinia
October-December 2017, 13(52):627-633
DOI:10.4103/pm.pm_470_16  PMID:29200724
Objective: Parkinson's disease, a slowly progressive neurological disease, is associated with degeneration of the basal ganglia of the brain and a deficiency of the neurotransmitter dopamine. The main aspects of researches are the protection of normal neurons against degeneration. Fatty acids (FAs), the key structural elements of dietary lipids, are carboxylic straight chains and notable parameters in nutritional and industrial usefulness of a plant. Materials and Methods: Black cumin, a popular anti-inflammatory and antioxidant food seasoning, contains nonpolar constituents such as FAs which were extracted using hexane. Different fractions and subfractions were apt to cytoprotection against apoptosis and inflammation induced by 1-methyl-4-phenylpyridinium (MPP+) in rat pheochromocytoma cell line (PC12) as a neural cell death model. The experiment consisted of examination of cell viability assessment, mitochondrial membrane potential (MMP), caspase-3 and -9 activity, and measurement of cyclooxygenase (COX) activity. Results: MPP+ induced neurotoxicity in PC12 cells. Pretreatment with subfractions containing FA mixtures attenuated MPP+-mediated apoptosis partially dependent on the inhibition of caspase-3 and -9 activity and increasing the MMP. A mixture of linoleic acid, oleic acid, and palmitic acid also decreased the COX activity induced by MPP+ in PC12 cells. Conclusion: Our observation indicated that subtoxic concentration of FA from Nigella sativa may exert cytoprotective effects through their anti-apoptotic and anti-inflammation actions and could be regarded as a dietary supplement. Abbreviations used: ANOVA: Analysis of variance; Ca: Calcium; CDCl3: Chloroform; COX: Cyclooxygenase; DMSO: Dimethyl sulfoxide; EA: Elidic acid; EDTA: Ethylene diamine tetraacetic acid; ELISA: Enzyme Linked Immunosorbent Assay; ESI-MS: Electron spray mass spectroscopy; FAs: Fatty acids; FBS: Fetal bovine serum; GC: Gas chromatography; 1HNMR: Hydrogen nuclear magnetic resonance; LA: Linoleic acid; MPP+: 1-Methyl-4-phenylpyridinium; MPTP: 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine; MTT: 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide; N. sativa: Nigella sativa; OA: Oleic acid; PA: Palmitic acid; PBS: Phosphate buffer saline; PC12: Rat pheochromocytoma cell line; PD: Parkinson's disease; PDA: Photo diode array detector; PGE2: Prostaglandin E2; TLC: Thin layer chromatography; TMPD: N,N,N',N'-tetramethyl-p-phenylenediamine; USA: United states of America.
  3 2,785 214
Ligustilide relieves complete Freund's adjuvant-induced mechanical hyperalgesia through inhibiting the activation of spinal c-Jun N-terminal kinase/c-jun pathway in rats
Yi-Rui Wang, Hui Xu, Min Tao, Li-Hua Xu, Xin-Chun Fu
October-December 2017, 13(52):634-638
DOI:10.4103/pm.pm_546_16  PMID:29200725
Background: Ligustilide, an active ingredient in a traditional Chinese medicine, has anti-inflammatory and analgesic effects. The underlying mechanisms of the anti-inflammatory pain effects of ligustilide are not completely understood. Objective: The aim of this study to investigate whether ligustilide conducts its analgesic effects on the complete Freund's adjuvant (CFA)-induced inflammatory pain through regulating the c-Jun N-terminal kinase (JNK)/c-Jun pathway in the spinal cord. Materials and Methods: Paw withdrawal thresholds (PWTs) and paw withdrawal latencies (PWLs) were tested to examine the analgesic effect of ligustilide on CFA-induced inflammatory pain in rats. The change of spinal JNK/c-Jun activation was detected by western blotting after CFA injection with or without consecutive intrathecal ligustilide administration. After SP600125 (JNK inhibitor) was intrathecally injected in CFA rats, PWTs and PWLs were tested to investigate the change of ligustilide's analgesic effect. Results: Repeated intravenous injection of ligustilide could attenuate the pain hypersensitivity induced by CFA. CFA caused increased activation of spinal JNK/c-Jun, which could be inhibited by ligustilide administration. Intrathecal injection of JNK inhibitor inhibited the CFA-induced mechanical hyperalgesia. Conclusion: Ligustilide could inhibit the upregulation of spinal p-JNK/p-c-Jun caused by CFA, and the inhibition of JNK/c-Jun activation is closely related to its anti-mechanical hyperalgesia effect in inflammatory pain. Abbreviations used: CFA: Complete Freund's adjuvant, JNK: c-Jun N-terminal kinase, MAPK: Mitogen-activated protein kinase, PWT: Paw withdrawal threshold, PWL: Paw withdrawal latency.
  3 2,325 190
In vitro and In vivo activity of Myrsine africana on elastase inhibition and anti-wrinkle activity
Namrita Lall, Navneet Kishore, Bianca Fibrich, Isa Anina Lambrechts
October-December 2017, 13(52):583-589
DOI:10.4103/pm.pm_145_17  PMID:29200717
Background: Myrsine africana (MA) is a plant traditionally used in South Africa to treat various diseases. Objective: The ethanolic extract of MA, was used for in vitro and in vivo studies to determine its elastase inhibitory activity. Materials and Methods: MA and its isolated compound, myrsinoside B, were tested in vitro for their elastase inhibitory activity. The MA extract was also evaluated for mutagenicity using two strains of Salmonella typhimurium (TA 98 and TA 100), microbial count, metal analysis, and stability. In vivo studies included irritancy and wrinkle reduction trials using Visioscan and Visioface. Results: The leaf extract showed good elastase inhibition with a 50% inhibitory concentration (IC50) of 28.04 μg/ml. Myrsinoside B inhibited the elastase enzyme at an IC50of 4.68 ± 0.34 μg/ml. No colony growth observed during mutagenicity studies and it was concluded that MA ethanolic extract is a nonmutagen. MA extract was found to be a nonirritant during the patch test clinical trial. MA was found to contain negligible amounts of microorganisms and heavy metals. Gel cream containing MA crude extract was found to be stable for 2 years when kept at temperatures below 30°C. In clinical trials (in vivo), it was found that the test product containing 5% ethanolic extract of MA was effective in reducing wrinkles after application 2 times a day for 14 days and 28 days compared to the placebo aqueous cream. Conclusion: MA is effective in reducing the appearance of wrinkles. Abbreviations used: 4-NQO: 4-nitroquinoline, D14-BL: Baseline to day fourteen, D28-BL: Baseline to day twenty-eight, CFU: Colony forming units, IC50: 50% inhibitory concentration, MA: Myrsine africana, MOU: Measurement of uncertainty, NaCl: Sodium chloride, NaH2PO4.H2O: Sodium phosphate monobasic monohydrate, SEM: Standard error of the mean, TA 98: Salmonella typhimurium strain 98, TA 100: S. typhimurium strain 100, TLC: Thin layer chromatography, TMA: Total microbial activity, XVB salt: Vogel-Bonner medium E.
  3 2,769 216
α-Glucosidase inhibitory activity from ethyl acetate extract of Antidesma bunius (L.) Spreng stem bark containing triterpenoids
Marista Gilang Mauldina, Rani Sauriasari, Berna Elya
October-December 2017, 13(52):590-594
DOI:10.4103/pm.pm_25_17  PMID:29200718
Background: Buni (Antidesma bunius [L.] Spreng) has been used as a traditional antidiabetic agent in Asia. Objective: The mechanism of antidiabetic properties was studied in this study by determine its α-glucosidase inhibitory activity. Method: Inhibition of α-glucosidase was performed in all fraction of Buni stem bark with acarbose and miglitol as standards. The half maximal inhibitory concentration (IC50) value of acarbose and miglitol was 5.75 and 59.76 μg/mL respectively while ethyl acetate (EtOAc) fraction was the most active fraction with IC50of 19.33 μg/mL. Three isolates (B1, B2, and B3) were found in the EtOAc fraction and elucidated by infrared, 1hydrogen-nuclear magnetic resonance, 13carbon-nuclear magnetic resonance, and two-dimensional nuclear magnetic resonance. Result: The chemical structures of the isolates were identified by the spectrum then compared with literature which concluded that B1 is friedelin, B2 is β-sitosterol, and B3 is betulinic acid. Inhibition of the α-glucosidase assay showed IC50values of B1, B2, and B3 were 19.51, 49.85, and 18.49 μg/mL, respectively. Abbreviations used: IC50: Half maximal inhibitory concentration; H-NMR: Hydrogen-nuclear magnetic resonance; C-NMR: Carbon nuclear magnetic resonance; 2D-NMR: Two dimensional-nuclear magnetic resonance; EtOH: Ethanol; EtOAc: Ethyl acetate; MeOH: Methanol; CHCl3: Chloroform; DMSO: Dimethyl sulfoxide; EtF: Ethyl acetate fraction; Na2CO3: Sodium carbonate; IR: Infrared; TGR5: Transmembrane G protein-coupled receptor 5; EC50: Half maximal effective concentration
  3 2,804 228
Identification of hepatoprotective constituents in Limonium tetragonum and development of simultaneous analysis method using high-performance liquid chromatography
Jae Sun Lee, Yun Na Kim, Na-Hyun Kim, Jeong-Doo Heo, Min Hye Yang, Jung-Rae Rho, Eun Ju Jeong
October-December 2017, 13(52):535-541
DOI:10.4103/pm.pm_477_16  PMID:29200710
Background: Limonium tetragonum, a naturally salt-tolerant halophyte, has been studied recently and is of much interest to researchers due to its potent antioxidant and hepatoprotective activities. Objective: In the present study, we attempted to elucidate bioactive compounds from ethyl acetate (EtOAc) soluble fraction of L. tetragonum extract. Furthermore, the simultaneous analysis method of bioactive EtOAc fraction of L. tetragonum has been developed using high-performance liquid chromatography (HPLC). Materials and Methods: Thirteen compounds have been successfully isolated from EtOAc fraction of L. tetragonum, and the structures of 1–13 were elucidated by extensive one-dimensional and two-dimensional spectroscopic methods including 1H-NMR, 13C-NMR, 1H-1H COSY, heteronuclear single quantum coherence, heteronuclear multiple bond correlation, and nuclear Overhauser effect spectroscopy. Hepatoprotection of the isolated compounds against liver fibrosis was evaluated by measuring inhibition on hepatic stellate cells (HSCs) undergoing proliferation. Results: Compounds 1–13 were identified as gallincin (1), apigenin-3-O-β-D-galactopyranoside (2), quercetin (3), quercetin-3-O-β-D-galactopyranoside (4), (−)-epigallocatechin (5), (−)-epigallocatechin-3-gallate (6), (−)-epigallocatechin-3-(3″-O-methyl) gallate (7), myricetin-3-O-β-D-galactopyranoside (8), myricetin-3-O-(6″-O-galloyl)-β-D-galactopyranoside (9), myricetin-3-O-α-L-rhamnopyranoside (10), myricetin-3-O-(2″-O-galloyl)-α-L-rhamnopyranoside (11), myricetin-3-O-(3″-O-galloyl)-α-L-rhamnopyranoside (12), and myricetin-3-O-α-L-arabinopyranoside (13), respectively. All compounds except for 4, 8, and 10 are reported for the first time from this plant. Conclusion: Myricetin glycosides which possess galloyl substituent (9, 11, and 12) showed most potent inhibitory effects on the proliferation of HSCs. Abbreviations used: HSQC: Heteronuclear single quantum coherence; HMBC: Heteronuclear multiple bond correlation; NOESY: Nuclear Overhauser effect spectroscopy; EGCG: Epigallocatechin-3-gallate; EGC: Epigallocatechin; HSC: Hepatic stellate cell; MTT: 3-(4,5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide.
  3 3,773 285
Gastroprotective and ulcer healing effects of camel milk and urine in HCl/EtOH, non-steroidal anti-inflammatory drugs (indomethacin), and water-restraint stress-induced ulcer in rats
Zijuan Hu, Xiaoman Chang, Qing Pan, Kebin Gu, Patrick Nwabueze Okechukwu
October-December 2017, 13(52):559-565
DOI:10.4103/pm.pm_135_17  PMID:29200713
Background: Camel milk has been reportedly used to treat dropsy, jaundice, tuberculosis, and diabetes while camel urine is used to treat diarrhea and cancer. However, there is no scientific evidence on the antiulcer activity of camel milk and urine. Thus, the present is designed to investigate the gastroprotective and ulcer healing effect of camel milk and urine on experimentally induced gastric ulcer models in rats. Materials and Methods: The gastroprotective effect was investigated in HCl/EtOH, water-restraint stress (WRS) and non-steroidal anti-inflammatory drugs (indomethacin)-induced ulcer models while ulcer healing activity was investigated in indomethacin-induced ulcer model. Cimetidine (100 mg/kg) was used as a standard antiulcer drug. Results: Acute toxicity study done up to a dosage of 10 ml/kg of camel milk and urine showed no signs of toxicity and mortality among the rats, indicating the present dosage of 5 ml/kg is safe to be administered to the rats. In the HCl/EtOH model, oral administration of cimetidine (100 mg/kg), camel urine (5 ml/kg), and camel milk (5 ml/kg) significantly (P < 0.05) inhibited gastric lesions by 83.7, 60.5 and 100%, respectively. In the WRS-induced model, cimetidine, and camel urine showed an ulcer inhibition of 100% while camel milk showed an inhibition of 50%. Similarly, in the indomethacin-induced ulcer model, cimetidine, camel milk, and urine showed an ulcer inhibition of 100, 33.3, and 66.7%, respectively. In addition, camel milk and urine also showed a significant (P < 0.05) ulcer healing effect of 100% in indomethacin-induced ulcer model, with no ulcers observed as compared to that of cimetidine, which offers a healing effect of 60.5%. Conclusion: The antiulcer activity of camel milk and urine may be attributed to its cytoprotective mechanism and antioxidant properties. Abbreviations used: NSAIDs: Non-steroidal anti-inflammatory drugs, UI: Ulcer index, ANOVA: One-way analysis of variance, WRS: Water-restraint stress, ROS: Reactive oxygen species
  3 5,338 258
Antioxidant, acetylcholinesterase, butyrylcholinesterase, and α-glucosidase inhibitory activities of Corchorus depressus
Samina Afzal, Bashir Ahmad Chaudhry, Ashfaq Ahmad, Muhammad Uzair, Khurram Afzal
October-December 2017, 13(52):647-651
DOI:10.4103/pm.pm_95_17  PMID:29200727
Background: Corchorus depressus (Cd) commonly known as Boa-phalee belonging to the family Tiliaceae having 50 genera and 450 species. Cd is not among the studied medicinal agent despite its potential in ethnopharmacology. Objectives: The present study investigated antioxidant, acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and α-glucosidase inhibitory activities of Cd. The dichloromethane and methanolic extracts of the Cd were evaluated for biological activities such as antioxidant and enzyme inhibitory activities of AChE, BChE, and α-glucosidase. Materials and Methods: Antioxidant activity was evaluated by measuring free radical scavenging potential of Cd using 1,1-diphenyl-2-picrylhydrazyl. Enzyme inhibition activities were done by measuring optical density. Results: The methanol extract of roots of Cd showed potential free radical scavenging activity 99% at concentration 16.1 μg/ml. AChE was inhibited by aerial part of dichloromethane fraction by 46.07% ± 0.45% while dichloromethane extracts of roots of Cd possessed significant activity against BChE with 86% inhibition compared with standard drug Eserine at concentration 0.5 mg/ml. The dichloromethane extract of roots of Cd showed 79% inhibition against α-glucosidase enzyme activity with IC50 62.8 ± 1.5 μg/ml. Conclusion: These findings suggest Cd as useful therapeutic option as antioxidant and inhibition of AChE, BChE, and α-glucosidase activities. Abbreviations used: DPPH: 1,1-diphenyl-2-picrylhydrazyl, Cd: Corchorus depressus, AChE: Acetylcholinesterase, BChE: Butyrylcholinesterase, AD: Alzheimer's disease.
  3 2,938 193
A study of the anti-cancer effects of the hexane fraction of the methanol extract of forsythiae fructus
Se-Eun Lee, Chiyeon Lim, Soon-Cheol Ahn, Suin Cho
October-December 2017, 13(52):719-724
DOI:10.4103/0973-1296.211079  PMID:29200739
Background: Forsythiae Fructus (FF) is a well-known medicinal herb derived from the dried fruits of Forsythia suspensa (Thunb.) Vahl. (Oleaceae). Recently, bioactive compounds isolated from hydrophobic solvent fractions of FF have been reported to have anti-oxidant, antibacterial, and anti-cancer effects. Objective: Almost all herbal medicines are derived from water extracts, which suggests different extraction methods might enhance the practical efficacies of herbal medicines. In this study, the authors further investigated the most potential anti-cancer fraction, that is, the hexane fraction (FFH) of the methanol extract (FFM) of the dried fruits of Forsythia suspensa. Materials and Methods: FFH was investigated by measuring its effects on the viability and apoptotic death of PC-3 cells (a prostate cancer cell line), on the expression levels of Bcl-2, Bax, cytochrome c, procaspase-9, procaspase-3 and PARP, and caspase-3 activity. Results: FFH significantly accelerated apoptotic cell death and decreased the protein levels of Bcl-2, procaspase-9, and procaspase-3. Conclusion: FFH can act as a pro-oxidative agent and induce the apoptosis of prostate cancer cells. Abbreviations used: FF: Forsythiae Fructus; FFM: Methanol extract of Forsythiae Fructus; FFH: Hexane fraction of the methanol extract; DCFH-DA: 2',7'-dichlorodihydro-fluorescein diacetate.
  3 3,188 218
Induction of apoptosis by Tithonia diversifolia in human hepatoma cells
Min-Ren Lu, Huey-Lan Huang, Wen-Fei Chiou, Ray-Ling Huang
October-December 2017, 13(52):702-706
DOI:10.4103/0973-1296.218113  PMID:29200736
Background: Traditional Chinese herb Tithonia diversifolia, belonging to the Compositae family, has long been applied for the treatment of liver diseases. In recent years, many reports also indicated that it possesses hepato-protective, anti-inflammatory, and anti-cancer activities. Objective: In this study, we evaluated whether T. diversifolia is an effective therapy for hepatocellular carcinoma (HCC). Materials and Methods: Dry leaves of T. Diversifolia were first extracted in ethyl acetate, then further fractionated by different ratio of n-hexane-ethyl acetate (8:2→0:1) or methanol as fractions 1-6 (Td-F1 to Td-F6), respectively. We first showed that the ethyl acetate extracts of T. diversifolia leaves (Td-L-EA) exhibits growth inhibition on human hepatoma HepG2 cells. To further check the extracts-induced apoptosis, microscopic observation, fragmented chromosomal DNA electrophoresis, apoptotic DNA-detection ELISA assay, flow cytometry, and Western blot analysis were performed. Results: After isolating the effective fractions from Td-L-EA, we found strong cytotoxic effects of fraction-2 (Td-F2). By further analyzing the mechanisms of cytotoxic activities using microscopic observation, fragmented chromosomal DNA electrophoresis, apoptotic DNA-detection ELISA assay, and flow cytometry, we found that induction of apoptosis such as DNA fragmentation increased the apoptosis rate and the apoptosis sub-G1 populations in Td-F2-treated HepG2 cells. In addition, we also confirmed Td-F2-induced degradation of caspase-8, caspase-9, caspase-3, and caspase-3 substrate PARP. Besides, Td-F2 also increased the Bcl-2 proapoptotic family protein Bax expression. Conclusion: In short, our results clearly showed the induction of apoptosis by ethyl acetate extracts of T. diversifolia leaves in human hepatoma HepG2 cells, suggesting its potential application as an antitumor agent. Abbreviations used: T. diversifolia, Tithonia diversifolia; HCC, Hepatocellular carcinoma DMEM, Dulbecco's modified Eagle's medium; DMSO, dimethyl sulfoxide; HRP, horseradish peroxidase; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide; OD, optical density; SDS-PAGE, sodium dodecylsulfate-polyacrylamide gel electrophoresis; PARP, Poly (ADP-ribose) polymerase; PBS, phosphate buffered saline; PI, propidium iodide.
  2 2,367 174
Simultaneous determination of ten constituents in chaiqin qingning capsule by high-performance liquid chromatography coupled with triple-quadrupole mass spectrometry
Ting Yu Li, Xiao Kui Huo, Lu Zheng, Chao Wang, Hai Jian Cong, Ting Xiang, Lin Zhang, Bao Jing Zhang, Shan Shan Huang, Bin Wu, Xin Yu Li
October-December 2017, 13(52):566-570
DOI:10.4103/pm.pm_81_17  PMID:29200714
Background: Chaiqin Qingning Capsule (CQQNC) was a prescription of Traditional Chinese Medicine with the effects of clearing away heat and removing toxin, harmonizing the exterior and interior, it was widely used in Asian, for example, China and Japan, different batches of the raws materials and different processing time may be the vital factor which raised a challenge to control the quality of the CQQNC. Experimental Methods: In this experiment, a high-performance liquid chromatography-mass spectrometry/MS (HPLC-MS/MS) method was developed to simultaneously determine ten bioactive components for the quality control of CQQNC. Chromatographic separation was achieved using an XBridge BEH C18 column (150 mm × 4.6 mm, 2.5 μm) with a mobile phase composed of 10 mm aqueous ammonium acetate and acetonitrile using a gradient elution in 20 min. This study was conducted by multiple reaction monitoring mode through electrospray ionization resource with a negative ionization mode. Results: The established method was validated with good performance of precision, accuracy, stability, and reproducibility and was utilized to simultaneously quantify ten constituents of CQQNC obtained from seven different batches. Conclusion: It is the first time to report the rapid and simultaneous analysis of the ten compounds in CQQNC by HPLC-MS/MS and apply to determine 10 constituents in 7 batches of CQQNC bought from drug store in china. This method could be considered as good quality criteria to control the quality of CQQNC. Abbreviations used: CHM: Chinese herbal medicine; TCM: Traditional Chinese Medicine; CQQNC: Triple-quadrupole mass spectrometry Chaiqin Qingning Capsules; HPLC–MS/MS: High liquid chromatography equipped with tandem mass spectrometry; ESI: Electrospray ionization; DP: Declustering potential; CE: Collision energy; RSD: Relative standard deviation; LOD: Limit of detection; LOQ: Limit of quantity.
  2 2,779 174
Antitumor effects of ethanol extracts from Hyptis rhomboidea in H22 tumor-bearing mice
Hong-Xin Cui, Lu Tang, Fang-Rong Cheng, Ke Yuan
October-December 2017, 13(52):571-575
DOI:10.4103/pm.pm_314_16  PMID:29200715
Background: Research the antitumor effects of ethanol extracts from Hyptis rhomboidea in H22 tumor-bearing mice. At the fist-stage of the experiments, the research team took MTT method to measure the antitumor activity in vitro, then selected the most inhibitory tumor cell strain as the test object of antitumor activity in vivo, established three models of a solid tumor H22 liver cancer, ascites tumor, and immunodeficiency in male mice. From inflammatory factor, liver toxicity, in vivo antioxidant index to observe antitumor activity of ethanol extracts from H. rhomboidea. Materials and Methods: Hundred and twenty ICR male mice were used to establish three models of a solid tumor H22 liver cancer, ascites tumor, and immunodeficiency in male mice and models group of a solid tumor H22 liver cancer randomly divided into six groupshe normal control group, the model control group, the positive group (cyclophosphamide), the sample treated group (high - 1.300 g/kg, medium - 0.750 g/kg, low - 0.373 g/kg). The animals were sacrificed 15d after oral administration and tumors were taken out for the tumor weights and antitumor rates. Blood in eyeball was collected for the determination of aspartate transaminase, alanine transaminase, malondialdehyde (MDA), superoxide dismutase (SOD), interleukin (IL)-2, and tumor necrosis factor (TNF)-α in serum. Sections of tumor issue were prepared, and morphological changes in tumor tissue cells were observed using hematoxylin and eosin staining technique. Results: The results showed that ethanol extracts from H. rhomboidea have a certain inhibitory effect on the digestive tumor cells. In solid tumor model, the inhibitory rate is up to 68.84% of the high dose of treated group from H. rhomboidea, and H. rhomboidea could improve the immune organ index, decrease the concentration of TNF-α and IL-2 in serum. In ascites tumor model, H. rhomboidea could slow down weight gain in mice and prolong the survival time; in immunodeficiency model, H. rhomboidea could improve the serum TNF-α and, IL-2 levels, increase SOD activity, and reduce MDA content, so as to achieve antitumor effect. Conclusions: Ethanol extracts from H. rhomboidea have obvious antitumor activity in vivo and can improve a tumor-burdened mice inflammation factors, improve the survival quality of H22 tumor mice, and enhance immunity and antitumor activity. Abbreviations used: H. rhomboidea: Hyptis rhomboidea; AST: Aspartate transaminase; ALT: Alanine transaminase; MDA: Malondialdehyde; SOD: Superoxide dismutase; IL-2: Interleukin 2; TNF-α: Tumor Necrosis Factor-α; CTX: Cyclophosphamide.
  2 2,611 149
Helichrysetin induces DNA damage that triggers JNK-mediated apoptosis in Ca Ski cells
Ho Yen Fong, Sri Nurestri Abd Malek, Hui Shin Yee, Saiful Anuar Karsani
October-December 2017, 13(52):607-612
DOI:10.4103/pm.pm_53_17  PMID:29200721
Background: Cervical cancer has become one of the most common cancers in women and currently available treatment options for cervical cancer are very limited. Naturally occurring chalcones and its derivatives have been studied extensively as a potential anticancer agent in different types of cancer and helichrysetin is naturally occurring chalcone that possess potent antiproliferative activity toward human cancer cells. Materials and Methods: Inhibitory activity of helichrysetin was evaluated at different concentrations. Ability of helichrysetin to induce apoptosis and its relation with c-Jun N-terminal kinase (JNK)-mediated mechanism of apoptosis was assessed using flow cytometry and Western blotting. Results: Helichrysetin inhibited Ca Ski cells at half maximal inhibitory concentration 30.62 ± 0.38 μM. This compound has the ability to induce DNA damage, mitochondrial membrane disruption, and loss of cell membrane integrity. We have shown that apoptosis was induced through the activation of JNK-mediated apoptosis by DNA damage in the cells then triggering p53-downstream apoptotic pathway with increased expression of pro-apoptotic proteins, Bax and caspase 3, and suppression of Bcl-2 anti-apoptotic protein. DNA damage in the cells also caused phosphorylation of protein ataxia-telangiectasia mutated, an activator of DNA damage response. Conclusion: We conclude that helichrysetin can inhibit Ca Ski cells through DNA damage-induced JNK-mediated apoptotic pathway highlighting the potential of this compound as anticancer agent for cervical cancer. Abbreviations used: ATM: Ataxia-telangiectasia mutated, DAPI: 4',6-diamidino-2-phenylindole, DMSO: Dimethyl sulfoxide, FITC: Fluorescein isothiocyanate, IC50: Half maximal inhibitory concentration, JC1-5,5',6,6'-Tetrachloro: 1',3,3'-tetraethylbenzimidazolylcarbocyanine, iodide, JNK: c-Jun N-terminal kinase, MMP: Mitochondrial membrane potential, PBS: Phosphate-buffered saline, SRB: Sulforhodamine B, TUNEL: Terminal deoxynucleotidyl transferase dUTP nick labeling
  2 2,714 153
Brazilian cerrado Qualea grandiflora Mart. leaves exhibit antiplasmodial and trypanocidal activities In vitro
Thuany de Moura Cordeiro, Fabian Borghetti, Sarah C Caldas Oliveira, Izabela Marques Dourado Bastos, Jaime Martins de Santana, Philippe Grellier, Sébastien Charneau
October-December 2017, 13(52):668-672
DOI:10.4103/pm.pm_100_17  PMID:29200731
Background: The rapid spread of drug-resistant strains of protozoan parasites required the urgent need for new effective drugs. Natural products offer a variety of chemical structures, which make them a valuable source of lead compounds for the development of such new drugs. Cerrado is the second largest biome in Brazil and has the richest flora of all the world savannahs. We selected Qualea grandiflora, a plant species known for its proprieties in folk medicine and its antibacterial activity. Objective: However, its antiprotozoal activity was not yet explored. Materials and Methods: We investigated the activities of fractions from the ethyl acetate extract of Q. grandiflora leaves against human life forms of Plasmodium falciparum, Trypanosoma cruzi, and Trypanosoma brucei gambiense, and for its cytotoxicity upon the rat L6-myoblast cell line. Ten fractions were produced by ethyl acetate:hexane chromatography. Results and Conclusion: The fractions showed no cytotoxicity against L-6 cells (IC50 > 100 μg/mL) and no hemolysis propriety. Three fractions had a moderate activity against P. falciparum, anyone was active against T. cruzi but four fractions demonstrated a high activity against bloodstream forms of T. brucei gambiense (8.0< IC50 <15 μg/mL). Identification and characterization of the active compounds are currently under investigation. Abbreviations used: CQ: Chloroquine, DMSO: Dimethyl sulfoxide, HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, HMI: Modified Iscove's medium, IC50: Concentration inhibiting 50% of parasite growth, IC90: Concentration inhibiting 90% of parasite growth, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, RPMI: Roswell Park Memorial Institute, SD: Standard deviation, SI: Ratio of cytotoxicity to biological activity − TC50/IC50, TC50: Concentration causing 50% of cell growth inhibition, TC90: Concentration causing 90% of cell growth inhibition, TLC: Thin-layer chromatography
  2 2,535 201
Enzyme inhibitors cause multiple effects on accumulation of monoterpene indole alkaloids in Catharanthus roseus cambial meristematic cell cultures
Zhou Pengfei, Zhu Jianhua, Yu Rongmin, Zi Jiachen
October-December 2017, 13(52):732-737
DOI:10.4103/0973-1296.218121  PMID:29200741
Background: Enzyme inhibitors have been used for the clarification of biosynthesis of natural products. Catharanthus roseus cambial meristematic cell (CMC) culture has been established and proved to be a better monoterpeneindole alkaloid (MIA) producer than C. roseus dedifferentiated cell (DDC) culture. However, little is known about the inter-relationship of the MIA-biosynthetic genes with respect to their transcription. Objective: To clarify effects of alteration of one gene transcription on transcript levels of another genes in MIA-biosynthetic pathway, and how the accumulation of MIAs in CMCs are influenced by the alteration of their biosynthetic gene transcript levels. Materials and Methods: 3-Hydroxy-3-methylglutaryl-CoA reductase (HMGR) inhibitor lovastatin and 1-deoxy-D-xylulose 5-phosphate synthase (DXS) inhibitor clomazone were fed to C. roseus CMC cultures. The contents of MIAs were qualified by High Performance Liquid Chromatography and the transcript levels of the relevant genes were measured by qRT-PCR. Results: Lovastatin improved the accumulation of MIAs via increasing the transcription of their biosynthetic genes encoding DXS1, tryptonphan decarboxylase (TDC), loganic acid methyltransferase (LAMT), strictosidine synthase (STR), desacetoxyvindoline-4-hydroxylase (D4H) and ORCA3 (a jasmonate-responsive transcriptional regulator), whereas clomazone reduced the contents of MIAs and the mRNA levels of the corresponding genes. Conclusion: The biosynthesis of MIAs in C. roseus is is manipulated via a complex mechanism, the knowledge of which paves the way for rationally tuning metabolic flux to improve MIA production in C. roseus CMCs.
  2 2,520 180
Contribution of the glucosinolate fraction to the overall antioxidant potential, cytoprotection against oxidative insult and antimicrobial activity of Eruca sativa Mill. leaves extract
Maria Fernanda Taviano, Antonietta Melchini, Angela Filocamo, Chiara Costa, Stefania Catania, Roberto Raciti, Shikha Saha, Paul Needs, Giuseppe Giovanni Bisignano, Natalizia Miceli
October-December 2017, 13(52):738-743
DOI:10.4103/pm.pm_245_16  PMID:29200742
Background: Eruca sativa Mill. (Brassicaceae) is commonly utilized as an ingredient in salads and also as a folk remedy to treat various diseases. Objective: The objective of this study was to establish the contribution of the glucosinolate (GLS) fraction to the overall antioxidant, cytoprotection against oxidative insult and antimicrobial properties of the hydro-alcoholic extract of E. sativa leaves from Sicily (Italy), characterized phytochemically. Materials and Methods: The antioxidant activity was evaluated by different in vitro systems. The cytoprotective effect against hydrogen peroxide (H2O2)-induced oxidative stress was tested in human peripheral blood mononuclear cells (PBMCs). The antimicrobial potential against bacteria and fungi was assayed by standard methods. Results: E. sativa extract exhibited both radical scavenging (50% inhibitory concentration [IC50] 1.04 ± 0.04 mg/mL) and ferrous ions-chelating activity (IC50 0.327 ± 0.0032 mg/mL) and mild reducing power; the GLS fraction showed chelating ability only (IC50 0.225 ± 0.009 mg/mL). In the experimental model of H2O2-induced oxidative stress in human PBMCs, a significant cytoprotective effect and a suppression of reactive oxygen species production by both extract and GLS fraction were observed (P < 0.001). E. sativa extract displayed moderate antimicrobial activity against Gram-positive bacteria, and Staphylococcus aureus was the most sensitive strain (minimum inhibitory concentration 0.125 mg/mL), whereas the GLS fraction was not active. Conclusion: GLSs are not involved in the primary antioxidant activity of E. sativa leaf extract but they are, almost in part, responsible for its ferrous ion-chelating properties. Iron-chelating compounds in E. sativa extract may protect cells under conditions of oxidative stress, and GLSs might play a chief role in this effect. Abbreviations used: GLS: Glucosinolate; H2O2: Hydrogen peroxide; PBMCs: Peripheral blood mononuclear cells; IC50: 50% inhibitory concentration; MIC: Minimum inhibitory concentration.
  2 2,450 231
Biological activities and cytotoxicity of Eperua oleifera Ducke oil-resin
Fernanda Torlania Alves Gomes, Ana Paula de Araújo Boleti, Lidiam M Leandro, Diego Squinello, Ellen S. P. Aranha, Marne C Vasconcelos, Paul Cos, Valdir F Veiga, Emerson Silva Lima
October-December 2017, 13(52):542-552
DOI:10.4103/pm.pm_552_16  PMID:29200711
Background: The oil-resin of Eperua oleifera Ducke has been used in popular medicine similarly to the copaiba oil (Copaifera spp.). Objective: This study aimed to investigate the effects of the acid fraction of E. oleifera oil-resin (AFEOR) on cell proliferation, collagen production in human fibroblasts, inhibition of metalloproteinases, and cytotoxicity against tumor cell lines. Materials and Methods: Acid fraction of E. oleifera was fractionated in the ion exchange column chromatography. Cytotoxicity and genotoxicity were evaluated by Alamar Blue® and Cometa assay. The inhibition of metalloproteinases was performed by zymography and Western blotting. Results: The predominant acidic diterpenes in the AFEOR were copalic and hardwickiic acids. AFEOR caused morphology alteration and decrease of proliferation at concentrations higher than 5 μg/mL. It also caused significant collagen proliferation in fibroblasts. It showed cytotoxicity against tumoral and nontumoral cell lines, with IC50values ranging from 13 to 50 μg/mL, and a hemolytic activity with an IC50value of 38.29 μg/mL. AFEOR inhibited collagenase activity, with an IC50value of 46.64 μg/mL, and matrix metalloproteinase-2 (MMP)-2 and MMP-9 in HaCaT cells or MMP-1 expression in MRC-5 cells. AFEOR induced genotoxicity in MRC-5 cells with a DNA damage index between 40% and 60% when compared to the negative controls (0%–20%). Conclusion: For the first time, biological activities from oil-resin E. oleifera demonstrated ratifying somehow its popular use. Abbreviations used: AFEOR: Eperua oleifera oil-resin.
  1 3,545 284
Evaluating the feasibility of five candidate DNA Barcoding Loci for Philippine Lasianthus Jack (Lasiantheae: Rubiaceae)
Muhammad Jefte C Arshed, Marcos B Valdez, Grecebio Jonathan D Alejandro
October-December 2017, 13(52):553-558
DOI:10.4103/pm.pm_1_17  PMID:29200712
Introduction: The pantropical genus Lasianthus Jack is identified for high phenotypic plasticity making traditional taxonomic identification difficult. Having some members with important medicinal properties, a precise complimentary identification through DNA barcoding is needed for species delineation. Materials and Methods: In this study, 12 samples representing six Philippine Lasianthus species were used to determine the most efficient barcoding loci among the cpDNA markers (matK, rbcL, rps16, and trnT-F) and nrDNA (ITS) based on the criteria of universality, discriminatory power, and resolution of species. Results: The results revealed that ITS has the recommended primer universality, greatest interspecific divergences, and average resolution of species. Among the cpDNA markers, matK and rbcL are recommended but with minimal resolution of species. While trnT-F showed moderate interspecific variations and resolution of Lasianthus species, rps16 has the lowest interspecific divergence and resolution of species. Conclusion: Consequently, ITS is the potential ideal DNA barcode for Lasianthus species. Abbreviations used: ITS: Internal Transcribe Spacer, matK: maturase K, rbcL: ribulose-1,5-biphospahte-carboxylase, rps16: ribosomal protein 16 small subunit gene.
  1 3,063 207
Two new phenolic glycosides from the aerial part of Dryopteris erythrosora
Guijae Yoo, SeonJu Park, Heejung Yang, Xuan Nhiem Nguyen, Nanyoung Kim, Jun Hyung Park, Seung Hyun Kim
October-December 2017, 13(52):673-676
DOI:10.4103/pm.pm_326_16  PMID:29200732
Background: Dryopteris erythrosora (D.C. Eaton) Kuntze is a species of fern in the family of Dryopteridaceae, which is distributed throughout East Asia. The genus Dryopteris has been used as traditional medicine, especially to treat hepatitis and protect liver. However, only few studies of chemical constituents of D. erythrosora have been conducted so far. Objective: In this study, we investigated the phytochemical constituents of D. erythrosora. Materials and Methods: The 80% methanol extract of the aerial part of D. erythrosora was used for the isolation of phenolic compounds. The isolated compounds were elucidated by various spectroscopic methods including nuclear magnetic resonance and mass spectrometry. Results: The present phytochemical investigation on the aerial part of D. erythrosora led to the isolation of two new phenolic glycosides, 1 and 2, as well as nine known flavonoids including two flavones (3 and 4) and seven flavonols (5-11). Conclusion: In this study, two new phenolic glycosides together with nine known flavonoids were isolated from the aerial part of D. erythrosora. Among them, compounds 4, 8, and 11 were isolated for the first time in Dryopteridaceae family from the present investigation. These results helped us to enrich our understanding of the chemical constituents of D. erythrosora and to identify compounds 1 and 2 which could be potential chemotaxonomic markers for the species. Abbreviations used: HPLC: High-performance liquid chromatography; Q-TOF LC/MS: Quadrupole-time-of-flight liquid chromatography/mass spectrometry; NMR: Nuclear magnetic resonance; TMS: Tetramethylsilane
  - 2,258 152
Zerumbone suppresses angiogenesis in HepG2 cells through inhibition of matrix Metalloproteinase-9, vascular endothelial growth factor, and vascular endothelial growth factor receptor expressions
Nozlena Abdul Samad, Ahmad Bustamam Abdul, Heshu Sulaiman Rahman, Abdullah Rasedee, Tengku Azmi Tengku Ibrahim, Yeap Swee Keon
October-December 2017, 13(52):731-736
DOI:10.4103/0973-1296.224274  PMID:29491625
Context: Due to increase in the number of patients with impaired immunity, the incidence of liver cancer has increased considerably. Aims: The aim of this study is the investigation the in vitro anticancer effect of zerumbone (ZER) on hepatocellular carcinoma (HCC). Materials and Methods: The anticancer mechanism of ZER was determined by the rat aortic ring, human umbilical vein endothelial cells (HUVECs) proliferation, chorioallantoic membrane, cell migration, and proliferation inhibition assays. Results: Our results showed that ZER reduced tube formation by HUVECs effectively inhibits new blood vessel and tissue matrix formation. Western blot analysis revealed that ZER significantly (P < 0.05) decreased expression of molecular effectors of angiogenesis, the matrix metalloproteinase-9, vascular endothelial growth factor (VEGF), and VEGF receptor proteins. We found that ZER inhibited the proliferation and suppressed migration of HepG2 cell in dose-dependent manner. Statistical Analysis Used: Statistical analyses were performed according to the Statistical Package for Social Science (SPSS) version 17.0. The data were expressed as the mean ± standard deviation and analyzed using a one-way analysis of variance. A P < 0.05 was considered statistically significant. Conclusion: The study for the first time showed that ZER is an inhibitor angiogenesis, tumor growth, and spread, which is suggested to be the mechanisms for its anti-HCC effect. Abbreviations used: ZER: Zerumbone, MMP-9: Matrix metalloproteinase-9, VEGF: Vascular endothelial growth factor, VEGFR: Vascular endothelial growth factor receptor, HUVECs: Human umbilical vein endothelial cells, HCC: Hepatocellular carcinoma, HIFCS: Heat inactivated fetal calf serum, DMSO: Dimethyl sulfoxide, EDTA: Ethyldiaminetetraacetic acid, Ig: Immunoglobulin, CAM: Chorioallantoic membrane, HRP: Horseradish peroxidase, NIH: National Institutes of Health, MTT: Microtetrazolium, SPSS: Statistical Package for Social Science.
  - 3,372 112
Cerebroprotective actions of Triticum aestivum Linn Powder and Bauhinia purpurea Flower powder in surgically induced cerebral infraction in rats
Akula Annapurna, Thonangi C Vishala, Veera R Bitra, Deepthi Rapaka, Asmath Shaik
October-December 2017, 13(52):737-741
DOI:10.4103/0973-1296.224309  PMID:29491626
Objective: The prime objective of this study is to evaluate the cerebroprotective actions of Triticum aestivum (wheatgrass) powder and Bauhinia purpurea flower (dev kanchan) powder against the experimentally induced global ischemia reperfusion injury in rats. Materials and Methods: In the first phase of the studies, 1 h before the surgical procedure, the Wistar rats were orally served with varied doses of wheatgrass powder (5, 10, 30, and 100 μg/kg) and Bauhinia flower powder (30, 100, 200, and 300 μg/kg), respectively. The ischemia was induced by 30-min bilateral carotid artery occlusion in succession to reperfusion for 4 h. It was proved that the wheatgrass powder and Bauhinia flower powder yielded a significant, dose-dependent cerebroprotection in terms of reduction in cerebral infarct size when compared with the control group. Coming to the second phase of the studies, a certain potential dose of 10 μg/kg of wheatgrass and 200 μg/kg of Bauhinia flower powders was selected keeping the protective action in view, and the animals were treated for 15 days. Results: The major findings of the study are that wheatgrass and Bauhinia flower powders significantly augmented the magnitude of the antioxidant enzymes, viz., super oxide dismutase and catalase, and further reduced the levels of lipid peroxidation. Conclusions: The present study clearly showed that the wheatgrass powder and Bauhinia flower powder possess significant antioxidant properties that may act as a key ingredient in various ayurvedic preparations for the treatment of various diseases like cerebral ischemic reperfusion injury. Abbreviations used: BCAO: Bilateral Carotid Artery Occlusion, MCA: middle cerebral artery, ROS: reactive oxygen species, SCMC: Sodium carboxy methyl cellulose, p.o: Per oral route, T.T.C: Triphenyl tetrazolium chloride, MDA: Malondialdehyde, SOD: Super oxide dismutase.
  - 2,036 100
Identification of inhibitors against metastasis protein “Survivin:In silico discovery using virtual screening and molecular docking studies
Swechha Mishra, Sangeeta Singh
October-December 2017, 13(52):742-748
DOI:10.4103/0973-1296.224310  PMID:29491627
Background: In experimental therapy of cancer, survivin is considered to be one of the well-established targets. Studies have found that it is overexpression in most of the human tumors, but it is rarely found in normal tissues. It is having varied structural and functional role. It controls cell division and cellular stress response and also regulates metastasis and migration of cancerous cells. It has also been recognized as a biomarker which makes it unconventional drug target. In spite of being one of the centrally active components in metastasis and invasion, their clinical use is minimal. To increase the therapeutic efficiency of cancer and its various stages, it is important to survey novel reagents targeting the pathways and mechanism involving survivin. Objective: The aim of this study was to identify novel survivin inhibitor candidates using in silico screening. Materials and Methods: In this course of work, virtual screening on a dataset of natural compounds retrieved from ZINC and other libraries were performed. Comparative analysis of the protein was done by studying the binding affinity of inhibitors that are already available. The best interacting complex was set for molecular dynamics simulation for 25 ns to validate the stability of system. These molecules were checked for their toxicity and absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties using OSIRIS and pre-ADMET tools. Results: We discovered ten such candidates with better binding efficiency with survivin in comparison to marketed chemical against the same. Furthermore, these inhibitor candidates did not induce cell toxicity. Binding affinity of reference molecules was varied from −6.8 to −8.5 kcal/mol while that of top scoring compound ZINC00689728 is −9.3 kcal/mol binding energy. Good placement and strong bond formation of selected molecule was observed during course of work. It is also having permissible ADMET property. Conclusion: Considering all the parameters, the screened molecule can be considered as a potential lead compound for designing new drug against survivin. Further investigation and testing will be required to make it to the final stage. Abbreviations used: MD: Molecular dynamics, LogS: Aqueous solubility, Acceptor HB: Hydrogen bond acceptor, Donor HB: Donor hydrogen bond donor, ADMET: Absorption, distribution, metabolism, excretion, and toxicity, RCSB: Research Collaboratory for Structural Bioinformatics, OPLS: Optimized potentials for liquid simulations, RMSD: Root-mean-square deviation.
  - 2,028 166
Taxifolin inhibits 7,12-Dimethylbenz(a)anthracene-induced breast carcinogenesis by regulating AhR/CYP1A1 signaling pathway
Md Wasimul Haque, Shakti Prasad Pattanayak
October-December 2017, 13(52):749-755
DOI:10.4103/0973-1296.224311  PMID:29491628
Background: Breast cancer (BC), because of its invasive characteristics, is one of the most common and deadliest cancers among the female population around the world. Research has demonstrated that AhR signaling also plays a vital role in BC initiation and development as well. Therefore, blocking this pathway to natural interferences paves a new channel for the prevention of BC. Several natural compounds such as flavonoids possess the anticancer activities against different cancers. Objective: The present study has been designed to estimate the chemotherapeutic potential of taxifolin (TAX) against 7,12-dimethylbenz(a)anthracene (DMBA)-induced mammary carcinoma in Sprague-Dawley rats. Materials and Methods: Initially, the molecular docking analysis of AhR and cytochrome P450s (CYPs) (CYP1A1 and CYP1B1) was performed using MAESTRO tool, in an attempt to rationalize the activity of TAX, based on their CYP1-binding potential. The in vitro CYP1A1 activity was determined by luciferase assay with CYP1A1 substrate luciferin CEE. The in vivo analysis was performed by administrating TAX at 10, 20, 40 mg/kg BW for 28 days intragastrically in DMBA induced (25 mg/animal dose) at 55 days of age Sprague-Dawley (SD) rats. BC initiates after 90 days of tumor induction phase. The molecular mechanism of TAX on Ahr and CYPs was also examined through the mRNA and protein expressions using reverse transcription-quantitative polymerase chain reaction and Western blotting analysis. Results: Furthermore, TAX altered the energy regulation on DMBA-induced BC in SD rats by considerably restoring the cancer-induced modulations in tumor growth. Our results showed that TAX reduced the expressions of CYP1A1 and CYP1B1 in DMBA-induced mammary carcinoma by downregulating the AhR signaling pathway. Conclusion: This study revealed that TAX might be able to act as a chemotherapeutic agent against CYP1A1- and CYP1B1-mediated cancer and the inhibition of the DMBA-induced mammary carcinogenesis in a rat model. Abbreviations used: CYPs: Cytochrome P450s; PAH: polycyclic aromatic hydrocarbons; HRP- Horseradish peroxidase; BSA: Bovine serum albumin; DTTP: Deoxythymidine Triphosphate (nucleotide); RT-qPCR: Real Time quantitative polymerase chain reaction; CADD: Computer Aided Drug Drafting.
  - 1,808 104
In vitro Cytotoxicity and apoptotic assay in HT-29 cell line using Ficus hispida Linn: Leaves extract
Jayalalitha Sathiyamoorthy, Natarajan Sudhakar
October-December 2017, 13(52):756-761
DOI:10.4103/0973-1296.224312  PMID:29491629
Background: Ficus hispida Linn. (Family Moraceae), well-known beneficial medicinal shrub, has been traditionally used for the treatment of various diseases such as leukoderma. Objective: The aim of the present study is to investigate the efficacy of F. hispida ethanolic leaves extract for antiproliferative, apoptotic, cell cycle blockade, and wound healing. Materials and Methods: F. hispida leaves extract was treated with colorectal adenocarcinoma cancer cell line HT29 for 24 h with control. The cells were treated at varying concentration ranges of 15, 31, 62, 125, and 250 μg/ml each The cytotoxicity effect of leaves extract was studied by 3-[4, 5-dimethylthiazol-2-yl]-2, 5 diphenyltetrazolium bromide assay and their anticancer activity was further evaluated using cell cycle analysis and wound scratch assay. Results: The end antiproliferative result showed that HT-29 cell viability decreases in a concentration-dependent manner and the growth inhibitory effect (IC50) values are obtained at a concentration of 125 μg. The increase in number of apoptotic cell was observed after treating HT-29 cells with the sample in double-staining methods. G0/G1 phase of the cell cycle was significantly blocked by the test sample followed by the G2/M phase in a negligible manner. In vitro cell wound closure or contracture was not significant when compared the sample against control group. Conclusion: F. hispida Linn. ethanolic leaves extract had shown to possess excellent cytotoxic effect through inducing apoptosis, especially causing cell cycle arrest at the G0/G1 phase. Abbreviations used: HT 29: Human adenocarcinoma colorectal cell line; PBS: Phosphate Buffered Saline; FBS: Fetal Bovine Serum; DMEM: Dulbecco's Modified Eagles Medium; MTT: 3 [4, 5 dimethylthiazol 2 yl] 2, 5 diphenyltetrazolium bromide; NCCS: National Centre for Cell Sciences; DMSO: DiMethyl SulfOxide; PI: Propidium Iodide; AO: Acridine Orange;EB: Ethidium Bromide; IC: Inhibitory Concentration.
  - 2,126 124
Safety profile investigations of Meyna spinosa (Roxb.) and Oroxylum indicum (Linn.) Extracts collected from Northeast India
Shweta Singh, Pronobesh Chattopadhyay, Sahindra Kumar Borthakur, Rudragoud Policegoudra
October-December 2017, 13(52):762-768
DOI:10.4103/0973-1296.224313  PMID:29491630
Background: Meyna spinosa (M.S) (Roxb.) ex Link and Oroxylum indicum (O.I) (Linn.) Vent, widely used traditional Northeast Indian medicinal plant used for various purposes, have not yet explored for safety profile. Objective: To investigate the safety profile of M.S (Roxb.) ex Link leaves and O.I (Linn.) Vent stem bark extracts collected from Northeast region of India. Materials and Methods: In this study, mutagenic, cytotoxic, and genotoxic and/or nontoxic potential of these two plant extracts using various toxicological investigations, as per the regulatory test guidelines, were evaluated. The mutagenic, cytotoxic, and genotoxic potential of these two plants were assayed using Ames test, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, comet assay, and micronucleus test in the bone marrow cells. Results: The results demonstrated that the tested doses of M.S (Roxb.) ex Link leaves extract showed mutagenic, cytotoxic, and genotoxic effects, whereas O.I (Linn.) Vent stem bark extracts showed nonmutagenic, noncytotoxic, and nongenotoxic effects. Conclusion: The stem bark extracts of O.I (Linn.) Vent has no mutagenic, cytotoxic, and genotoxic or clastogenic effects in our experimental conditions. However, M.S (Roxb.) ex Link leaves extract caused a significant increase in DNA damage as compared with the positive control, i.e., cyclophosphamide. Thus, the present study revealed that M.S (Roxb.) ex Link leaves extract is toxic, while O.I (Linn.) Vent stem bark extract was found to be safe. Abbreviations used: MS: Meyna spinosa; OI: Oroxylum indicum.
  - 4,697 99
Black Tea (Camellia sinensis) Extract induced prenatal and postnatal toxicity in experimental albino rats
Avijit Dey, Antony Gomes, Subir Chandra Dasgupta
October-December 2017, 13(52):769-774
DOI:10.4103/pm.pm_141_17  PMID:29491631
Background: Tea (Camellia sinensis) being the most widely drank beverage and despite having numerous beneficial role toward health and disease, its safety evaluation during pregnancy and prenatal, postnatal developmental period need to be monitored. Objective: This study was to evaluate the toxicity of black tea extract (BTE) in experimental pregnant rats and on their pups during prenatal and postnatal developmental periods. Materials and Methods: Pregnant female (120 ± 10 g) Wister albino rats were chosen for this study. Group 1 was control group where pregnant female rats were treated with saline. Group 2 and Group 3 were pregnant female rats treated with 50 mg and 100 mg BTE/kg/day, respectively, throughout prenatal and postnatal periods. All three groups of rats were provided food and drinking water ad libitum. Animals were examined through their urinary and serum parameters, histopathological studies, and biomorphometric studies in pups. All data were expressed as mean ± standard deviation with significance between the controls and the treated groups (n = 6). Collected data were subjected to the analysis of variance and Tukey test; P < 0.05 was considered as statistically significant. Results: BTE produced significant alterations in urinary calcium, creatinine, and urea during prenatal period; exhibited proteinuria, ketonuria, and histology showed nephrotoxicity during postnatal period, and BTE also showed a significant increase in serum proinflammatory cytokines and decreased anti-inflammatory cytokines level compared to control group. BTE caused significant changes in biomorphometric parameters in the pups as compared with pups of control mothers. Conclusion: This study confirmed the BTE-induced toxicity in pregnant rats and their pups. Abbreviations used: BTE: Black tea extract, IL-1α: Interleukin 1 alpha, IL-1 β: Interleukin 1 beta, IL-6: Interleukin 6, IL-10: Interleukin 10, TNF-α: Tumor necrosis factor alpha.
  - 1,603 88
Biochemical characterization of fungus isolated during In vitro Propagation of Bambusa balcooa
Bhawna Tyagi, Salil Tewari, Ashutosh Dubey
October-December 2017, 13(52):775-779
DOI:10.4103/0973-1296.224315  PMID:29491632
Background: Bambusa balcooa (Poaceae: Bambusoideae) is a multipurpose bamboo species, which is native of the Indian subcontinent. B. balcooa is regarded as one of the best species for scaffolding and building purposes because of its strong culm. Other uses include paper pulp, handicrafts, and products of the wood chip industry. Due to these various uses in industries, this species has been identified as one of the priority bamboos by the National Bamboo Mission. Objective: This study is designed to analyze the identification of fungus and develop the strategy to eliminate the contamination during in vitro establishment of B. balcooa through nodal part. Fungus contamination is a problem which is encountered during in vitro establishment of B. balcooa cultures. Materials and Methods: In the present study, fungus contamination from in vitro cultured plant has been isolated and subjected to partial sequence analysis of the 18S rRNA gene to identify the fungus strain. Experiments were designed to develop a strategy for removal of the fungus contamination with the help of antifungal compounds and commercial antimicrobial supplement supplied by HiMedia. Results: Fusarium equiseti was identified as endophytic fungus. It was observed that antimicrobial supplement at concentration of 500 μl/l was more effective concentration to remove fungus contamination and not showed any detrimental effect on growth parameters of shoot. Conclusion: This experiment would help in identification and to get rid of fungal contamination and improve the in vitro establishment of B. balcooa cultures for large-scale propagation. Abbreviations used: B. balcooa: Bambusa balcooa, F. equiseti: Fusarium equiseti, PDA: Potato dextrose agar, PCR: Polymerase chain reaction, MS: Murashige and Skoog's, BAP: 6-Benzylaminopurine, ITS1/4: Internal transcribed spacer region 1/4, GA3: Gibberellic acid
  - 1,401 94
Computational breakthrough of natural lead hits from the genus of Arisaema against human respiratory syncytial virus
Kamal Kant, Uma Ranjan Lal, Manik Ghosh
October-December 2017, 13(52):780-785
DOI:10.4103/0973-1296.224316  PMID:29491633
Background: To date, efforts for the prevention and treatment of human respiratory syncytial virus (RSV) infection have been still vain, and there is no safe and effective clinical accepted vaccine. Arisaema genus has claimed for various traditional bioactivities, but scientific assessments are quite limited. Objective: This encouraged us to carry out our present study on around 60 phytoconstituents of different Arisaema species as a natural inhibitor against the human RSV. Materials and Methods: Selected 60 phytochemical entities were evaluated on the docking behavior of human RSV receptor (PDB: 4UCC) using Maestro 9.3 (Schrödinger, LLC, Cambridge, USA). Furthermore, kinetic properties and toxicity nature of top graded ligands were analyzed through QikProp and ProTox tools. Results: Notably, rutin (glide score: −8.49), schaftoside (glide score: −8.18) and apigenin-6,8-di-C-β-D-galactoside (glide score − 7.29) have resulted in hopeful natural lead hits with an ideal range of kinetic descriptors values. ProTox tool (oral rodent toxicity) has resulted in likely toxicity targets of apex-graded tested ligands. Conclusion: Finally, the whole efforts can be explored further as a model to confirm its anti-human RSV potential with wet laboratory experiments. Abbreviations used: RSV: Respiratory syncytial virus, PRRSV: Porcine respiratory and reproductive syndrome virus, ADME-T: Absorption, distribution, metabolism, excretion, and toxicity.
  - 1,473 105
In silico prediction and wet lab validation of Arisaema tortuosum (Wall.) schott extracts as antioxidant and anti-breast cancer source: A comparative study
Kamal Kant, Uma Ranjan Lal, Manik Ghosh
October-December 2017, 13(52):786-790
DOI:10.4103/0973-1296.224317  PMID:29491634
Background: Globally, reactive oxygen species have served as an alarm predecessor toward pathogenesis of copious oxidative stress-related diseases. The researchers have turned their attention toward plant-derived herbal goods due to their promising therapeutic applications with minimal side effects. Arisaema tortuosum (Wall.) Schott (ATWS) is used in the traditional medicine since ancient years, but scientific assessments are relatively inadequate and need to be unlocked. Objective: Our aim was designed to validate the ATWS tuber and leaf extracts as an inhibitor of oxidative stress using computational approach. Materials and Methods: The reported chief chemical entities of ATWS were docked using Maestro 9.3 (Schrödinger, LLC, Cambridge, USA) tool and further ATWS extracts (tubers and leaves) were validated with 2,2'-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), ferric-reducing ability of plasma (FRAP), and sulforhodamine B assays experimentally. Results: In silico results showed notable binding affinity of ATWS phytoconstituents with the receptor (PDB: 3ERT). Experimentally, butanolic tuber fraction confirmed promising antioxidant potential (ABTS: IC50: 271.67 μg/ml; DPPH: IC50: 723.41 μg/ml) with a noteworthy amount of FRAP (195.96 μg/mg), total phenolic content (0.087 μg/mg), and total flavonoid content (7.5 μg/mg) while chloroform fraction (leaves) showed considerable reduction in the cell viability of MCF-7 cell line. Conclusion: The current findings may act as a precious tool to further unlock novel potential therapeutic agents against oxidative stress. Abbreviations used: ATWS: Arisaema tortuosum (Wall.) Schott, DPPH: 2,2'-diphenyl-1-picrylhydrazyl, ABTS: 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, FRAP: Ferric-reducing ability of plasma, TPC: Total phenolic content, TFC: Total flavonoid content, SRB: Sulforhodamine B
  - 1,473 102
Phytochemical study of Aegle marmelos: Chromatographic elucidation of polyphenolics and assessment of antioxidant and cytotoxic potential
Farina Mujeeb, Ahamad Faiz Khan, Preeti Bajpai, Neelam Pathak
October-December 2017, 13(52):791-800
DOI:10.4103/0973-1296.224321  PMID:29491635
Background: The antioxidant potential of medicinal plants has been illustrated through many reports clearly depicting that plants are a rich source of antioxidants, making them a great resource of novel drugs and health-care products. Objectives: The current study is, therefore, focused toward the assessment of antioxidant properties along with the presence of phytochemicals in leaves of 18 varieties/accessions of Aegle marmelos. Materials and Methods: The antioxidant activities were initially measured using superoxide radical scavenging method, 2, 2-diphenyl-1-picrylhydrazyl (DPPH), and ferric-reducing ability of plasma assays. Further, thin-layer chromatography (TLC), high-performance TLC, and column chromatography were performed to isolate the potentially active fraction and anti-inflammatory activity of crude, and the isolated fraction was tested on J774 macrophage cell line. Results: The maximum inhibition of superoxide anions was shown by Pant Aparna. Additionally, Pant Aparna extract was most efficient, exhibiting 92.0% inhibition in scavenging the DPPH radicals. The content of total carotenoids was found to be higher in Pant Aparna among all the varieties/accessions. Furthermore, the crude extract and the fraction A. marmelos methanolic fraction 21 (AMMF21) were found to be nontoxic and significant reactive oxygen species, and NO inhibition was observed in a concentration-dependent manner. Moreover, the methanolic extract of variety Pant Aparna showed promising in vitro antioxidant activity, indicating its potency for therapeutic applications. Conclusion: In brief, this is the first ever report on Pant Aparna as the best variety in terms of phytocompounds and identification of potential antioxidant activity. In addition, the AMMF21 fraction of methanolic extract possessing best antioxidant activity on macrophage cells indicates its use as a novel phytotherapeutic agent. Abbreviations used: AMMF21: Aegle marmelos methanolic fraction 21, DPPH: (2, 2-diphenyl-1-picrylhydrazyl), FRAP: Ferric-reducing ability of plasma, HP-TLC: High-performance-thin-layer chromatography, TLC: Thin-layer chromatography, TCA: Trichloroacetic acid, TPTZ: 2,4,6-Tripyridyl-s-triazine, DNPH: 2,4-dinitrophenyl hydrazine, NBT: Nitroblue tetrazolium, NADH: Nicotinamide adenine dinucleotide, PMS: Phenazine metho-sulfate, DMEM: Dulbecco's modified Eagle medium; MTT: (3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide, DCFDA: 2',7'-dichlorofluorescein diacetate, LPS: Lipopolysaccharide, NED: N-(1-Naphthyl) ethylenediamine.
  - 2,069 126
Viwithan, a Standardized Withania somnifera root extract induces apoptosis in murine melanoma cells
HV Sudeep, K Gouthamchandra, BJ Venkatesh, K Shyam Prasad
October-December 2017, 13(52):801-806
DOI:10.4103/0973-1296.224324  PMID:29491636
Background: Withania somnifera is an Indian medicinal herb known for the multipotential ability to cure various therapeutic ailments as described in the ayurvedic system of medicine. Objective: In the present study, we have evaluated the antiproliferative activity of a standardized W. somnifera root extract (Viwithan) against different human and murine cancer cell lines. Materials and Methods: The cytotoxicity of Viwithan was determined using thiazolyl blue tetrazolium blue assay and crystal violet staining. The apoptotic changes in B16F1 cells following treatment with Viwithan were observed by acridine orange/ethidium bromide (AO/EB) staining and DNA fragmentation assay. The binding affinity of withanolides in Viwithan with antiapoptotic proteins B-cell lymphoma 2, B-cell lymphoma-extra large, and myeloid cell leukemia 1 (MCL-1) were studied using in silico approach. Results: The half maximal inhibitory concentration (IC50) values of Viwithan against liver hepatocellular carcinoma, Henrietta Lacks cervical carcinoma cells, human colorectal carcinoma cell line, and Ehrlich ascites carcinoma cells were 1830, 968, 2715, and 633 μg/ml, respectively. Interestingly, Viwithan was highly effective against B16F1 cells with an IC50 value of 220 μg/ml after 24 h treatment. The morphological alterations of apoptotic cell death were clearly observed in the AO/EB-stained cells after treatment with Viwithan. Viwithan induced late apoptotic changes in treated B16F1 cells as evident by the ladder formation of fragmented DNA in a time-dependent manner. The findings of molecular docking showed that withanolides present in Viwithan have a more binding affinity with the antiapoptotic proteins, particularly MCL-1. Conclusion: We have reported for the first time that Viwithan with 5% withanolides has a potent cytotoxic effect, particularly against B16F1 murine melanoma cells among the different cancer cell lines tested. Abbreviations used: MTT: Thiazolyl blue tetrazolium blue; DMSO: Dimethyl sulfoxide; BSA: Bovine serum albumin; DMEM: Dulbecco's minimum essential medium; NCCS: National Centre for Cell Science; PBS: Phosphate-Buffered Saline; HepG2: Liver hepatocellular carcinoma; HeLa: Henrietta Lacks cervical carcinoma cells; HCT-116: Human colorectal carcinoma cell line; EAC: Ehrlich ascites carcinoma cells; IC50: Half maximal inhibitory concentration; AO/EB: Acridine orange/Ethidium bromide; BCL-2: B-cell lymphoma 2; BCL-XL: B-cell lymphoma-extra large; MCL-1: Myeloid cell leukemia 1; PDB: Protein Data Bank; ANOVA: Analysis of variance.
  - 1,535 90
Protective effect of Exacum lawii on cisplatin-induced oxidative renal damage in rats
Sonam Sharma, Arusha Modi, Gopeshwar Narayan, Siva Hemalatha
October-December 2017, 13(52):807-816
DOI:10.4103/0973-1296.224325  PMID:29491637
Background: Exacum lawii (Gentianaceae), a bitter herb conventionally used in kidney diseases and eye problems, endemic to the Western coast and Southern part of India. Aim: Folklore reports encourage the author to explore the nephroprotective effect of the standardized ethanolic extract of E. lawii against cisplatin-induced renal toxicity in the rat to scientifically validate its traditional use. Materials and Methods: Ethanolic extract of the whole plant of E. lawii was standardized with swertiamarin (secoiridoid glycoside) using high-performance liquid chromatography and tested for subacute toxicity according to the OECD guidelines. Nephroprotective potential at different doses of extract was evaluated against cisplatin (6 mg/kg, intraperitoneal) in experimental rats. The changes in serum renal toxicity markers, renal tissue oxidative stress biomarkers, and proinflammatory cytokines level were measured. To estimate the change in oxidative status of renal tissues, DNA and single viable cells were isolated from treated rat kidney, DNA fragmentation assay and flow cytometric analysis of reactive oxygen species (ROS) were performed. Histopathology of renal tissues was also examined. Results: Swertiamerin was found to be 119.59 mg/g of extract. Administration of E. lawii extract (ELE) restored the biochemical parameters. It also decreases the elevated proinflammatory cytokines level in kidney tissues and protected rat kidneys from oxidative stress in rats. Nephroprotective activity was validated by estimating ROS production in kidney live cells and DNA damage in kidney tissue. The histological architecture was also conserved. Conclusion: ELE showed significant renal protection against cisplatin through reducing oxidative stress and inflammation. Abbreviations used: ELE: Exacum lawii ethanolic extract; WHO: World Health Organization; SOD: Superoxide dismutase; CAT: Catalase; MDA: Malondialdehyde; HPTLC: High performance thin layer chromatography; p.o.: Per oral; i.p.: Intraperitoneal; TNF-α: Tumor necrosis factor alpha; IL-1β: Interleukin 1 beta; IL-6: Interleukin 6; ROS: Reactive oxygen species.
  - 2,096 97
Rhinacanthins-rich extract enhances glucose uptake and inhibits adipogenesis in 3T3-L1 Adipocytes and L6 Myotubes
Muhammad Ajmal Shah, Chanawee Jakkawanpitak, Decha Sermwittayawong, Pharkphoom Panichayupakaranant
October-December 2017, 13(52):817-821
DOI:10.4103/0973-1296.224326  PMID:29491638
Background: Obesity is one of the imperative dynamics in the incidence and intensification of type 2 diabetes mellitus (T2DM). Rhinacanthus nasutus leaf extracts are previously reported for their antidiabetic and antiobesity potential. Objective: The present study was performed to evaluate glucose uptake stimulatory and antiadipogenic activities of a standardized rhinacanthins-rich extract (RRE) and its marker compounds namely rhinacanthin-C (RC), rhinacanthin-D (RD), and rhinacanthin-N (RN) in 3T3-L1 and L6 cells. Materials and Methods: RRE was prepared by a green extraction process, and the marker compounds (RC, RD, and RN) were isolated from the RRE using a silica gel column chromatography. Glucose uptake stimulation in both 3T3-L1 and L6 cells was performed by quantification of residual glucose in the media using glucose oxidase kit. Antiadipogenic activity in 3T3-L1 adipocytes was performed by intracellular lipids quantification using oil red O dye. Results: At the highest effective dose, RRE (20 μg/mL) exhibited satisfactory glucose uptake stimulatory effect in 3T3-L1 adipocytes that equivalent to RN (20 μg/mL) and the positive control insulin (0.58 μg/mL) but higher than RC (20 μg/mL) and RD (20 μg/mL). In addition, treatments of L6 myotubes showed that RRE (2.5 μg/mL) exhibited potent and equivalent glucose uptake stimulation (>80%) to RC (2.5 μg/mL) and the standard drugs, insulin (2.90 μg/mL) and metformin (219.5 μg/mL), but higher than RD (2.5 μg/mL) and RN (2.5 μg/mL). Furthermore, RRE (20 μg/mL) exhibited potent antiadipogenic effect in 3T3-L1 adipocytes, which equivalent to RC (20 μg/mL) but higher than RD (20 μg/mL) and RN (20 μg/mL). Conclusions: The undertaken study suggests that RRE could be used as an effective remedy in the treatment of obesity-associated T2DM. Abbreviations used: T2DM: Type-2 diabetes mellitus; RRE: Rhinacanthins-rich extract; RC: Rhinacanthin-C; RD: Rhinacanthin-D; RN: Rhinacanthin-N; α-MEM: α-Minimum essential medium; DMEM: Dulbecco's modified Eagle's medium; HS: Horse serum; FBS: Fetal bovine serum; BSA: Bovine serum albumin; IBMX: 3-isobutyl-1-methylxanthine; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; GO: Glucose oxidase; NMR: Nuclear magnetic resonance; HPLC: High-performance liquid chromatography.
  - 1,632 123
Expression of hepatic cytochrome P450s in rats administered with Guibi-tang, a traditional herbal formula
Seong Eun Jin, Hyekyung Ha, Chang-Seob Seo, Hyeun-Kyoo Shin, Soo-Jin Jeong
October-December 2017, 13(52):822-827
DOI:10.4103/0973-1296.224327  PMID:29491639
Objective: The aim of this study was to investigate the possible herb-drug interactions between the traditional herbal formula Guibi-tang (GBT; Guipi-tang, Kihi-to) and conventional drugs. Materials and Methods: GBT was orally administered to either male or female Sprague Dawley (SD) rats once daily at doses of 1000, 2000, or 5000 mg/kg/day for 13 weeks. The messenger ribonucleic acid (mRNA) expression of drug-metabolizing enzyme cytochrome P450 isozymes (cytochrome P450s; CYP1A1, 1A2, 2B1/2, 2C11, 2E1, 3A1, 3A2, and 4A1) was analyzed in hepatic tissues by reverse transcription-polymerase chain reaction. Results: Repeated oral administration of GBT did not significantly influence the mRNA expression of hepatic CYP1A1, 1A2, 2B1/2, 2C11, 2E1, 3A1, 3A2, and 4A1 in male rats. By contrast, in female rats, the mRNA expression of hepatic CYP1A2 and 2B1/2 was significantly increased by repeated GBT treatment. Conclusion: Our findings indicate that caution is required in females when GBT is taken concomitantly with conventional drugs metabolized by CYP1A2 or 2B1/2. Our results provide information regarding the safety and effectiveness of GBT for clinical use. Abbreviations used: CYP450: Cytochrome P450s, GBT: Guibi-tang, SD: Sprague Dawley, HPLC: High-performance liquid chromatography, OECD: Organization for Economic Cooperation and Development, RNA: Ribonucleic acid, RT-PCR: Reverse transcription-polymerase chain reaction, GADPH: Glyceraldehyde-3-phosphate dehydrogenase.
  - 1,431 99
Antibacterial synergy of silver nanoparticles with gentamicin and chloramphenicol against Enterococcus faecalis
Sagar Katva, Satyajeet Das, Harpreet Singh Moti, Anupam Jyoti, Sanket Kaushik
October-December 2017, 13(52):828-833
DOI:10.4103/0973-1296.224328  PMID:29491640
Background: Enterococcus faecalis (Ef) is a multidrug-resistant pathogenic bacteria associated with hospital-acquired infections. Ef is involved in a number of infectious diseases. It generally infects patients with the weekend immune system, i.e. a person mostly acquires Ef infections in the hospital, especially in intensive care units and thus, is more likely to be resistant to many antibiotics. Development of resistance against various antibiotics and emergence of drug-resistant strains is a growing global concern. Objective: Due to the unselective use of antibiotics for a long time multidrug resistant bacteria and extensively drug-resistant, which is now posing a new challenge to the medical community. To treat infections caused by Ef, the synergistic effect of different antibiotics with silver nanoparticles (AgNPs) was tested against Ef. Materials and Methods: In the present study, synthesis of AgNPs was carried out from the cell-free supernatant of Klebsiella pneumoniae. AgNPs were characterized using various techniques, namely, ultraviolet-visible spectrophotometry, transmission electron microscopy, and Fourier transform infrared spectroscopy. Moreover, process optimization was done for enhanced production of AgNPs. In addition, antimicrobial activity of the nanoparticles was also tested. Furthermore, the nanoparticles were evaluated for their antimicrobial activities in combination with gentamicin and chloramphenicol, against Ef. Results: The results showed that the combination of gentamicin and chloramphenicol with AgNPs has a better antibacterial effect. To add to this, hemolytic activity of AgNPs was evaluated against human red blood corpuscles (RBCs). AgNPs were found to be nontoxic to RBCs. Conclusion: The collective effect of AgNps with Gentamicin and Chloramphenicol was more as compared to AgNps alone which indicate the synergistic effect of these components. These observations show the potential of AgNPs in combination with above-stated antibiotics against Ef infections. Abbreviations used: Ef: Enterococcus faecalis, MDR: Multidrug resistance, AgNPs: Silver nanoparticles, Kp: Klebsiella pneumoniae, RBCs: Red blood corpuscles, ENPs: Engineered nanoparticles, FTIR: Fourier transform infrared spectroscopy, TEM: Transmission electron microscopy, AgNO3: Silver nitrate, EDTA: Ethylenediaminetetraacetic acid, PBS: Phosphate-buffered saline.
  - 1,772 118
Triphala, regulates adipogenesis through modulation of expression of adipogenic genes in 3T3-L1 Cell line
Jyotibala Banjare, Prerna Raina, Prakash Mansara, Ruchika Kaul Ghanekar, Supriya Bhalerao
October-December 2017, 13(52):834-839
DOI:10.4103/0973-1296.224329  PMID:29491641
Background: Triphala, an Ayurvedic polyherbal formulation, is used for the treatment of various diseases including obesity. Objective: The present study was planned to evaluate the anti-adipogenic potential of aqueous extract of Triphala (TPaq) using 3T3-L1 adipocyte cell line model. Methods: The effect of aqueous extract of Triphala (TPaq) was tested on the viability of 3T3- L1 cells by MTT assay. The cells were treated with a cocktail of dexamethasone (DEX), isobutylmethylxanthine (IBMX) and insulin to induce adipogenesis. The cells were treated either with the induction cocktail or with the cocktail containing different concentrations (1, 10 and100 μg/ml) of TPaq. Intracellular lipid content was analyzed using Oil O Red stain and was quantified after extracting with isopropanol at 500 nm wavelength. The expression of early (PPAR-γ and C/EBP-α) and late (GLUT4 and FAS) phase adipogenic genes was studied by real time PCR. Results: TPaqdid not affect the viability of 3T3-L1 cell line. Interestingly, TPaqinduced a concentration dependant decrease in the intracellular lipid content and expression of both early and late phase adipogenic genes. This decrease was statistically significant compared to cells treated with only induction cocktail. Conclusion: These results suggested that Triphala regulated lipid accumulation by down regulating expression of adipogenic genes, resulting into prevention of adipogenesis. Abbreviations used: TPaq: Aqueous extract of Triphala; DMEM: Dulbecco's Modified Eagle's medium; FBS: Fetal Bovine Serum; IBMX: Isobutyl methylxanthine; DMX: Dexamethasone; MTT: [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide] assay; PPARγ: Peroxisome proliferator-activated receptor; C/EBP:Enhancer binding protein α, FAS:Fatty acid synthase; Glut-4: Glucose phosphate transporter 4.
  - 1,850 112
Green synthesis of silver nanoparticles using leaf extract of common arrowhead houseplant and its anticandidal activity
Mohammad Yasir, Jaspreet Singh, Manish Kumar Tripathi, Pushpendra Singh, Rahul Shrivastava
October-December 2017, 13(52):840-844
DOI:10.4103/0973-1296.224330  PMID:29491642
Background: Silver nanoparticles have excellent medical and nonmedical properties and application compared with other metallic nanoparticles. In the present study, fresh leaves of Syngonium podophyllum have been used for synthesis of silver nanoparticles. Objectives: In this study, we evaluated the anticandidal activity of S. podophyllum and the synthesized nanoparticles. Materials and Methods: In this study, simple and economical procedure was adopted for silver nanoparticles synthesis. S. podophyllum leaf was processed to obtain aqueous extract as a biological material for nanoparticles production. Synthesized nanoparticles were characterized by ultraviolet (UV) spectroscopy, X-ray diffraction (XRD), and atomic force microscopy. Results: The progress of silver nanoparticles biosynthesis from leaf extract of S. podophyllum was observed by UV-visible spectroscopy. The peaks maxima were observed at 455 nm for silver nanoparticles synthesized from the leaf extracts of S. podophyllum. XRD diffractogram showing Bragg peaks of face-centered cubic crystalline elemental silver confirming the formation of silver nanoparticles. The minimal inhibitory concentration values of aqueous extracts of S. podophyllum leaf were estimated by broth dilution method and found that the extracts exhibited antifungal activity against Candida albicans. The antifungal activity was also determined using disk diffusion method by measuring the diameter for zone of inhibition. Conclusion: S. podophyllum leaf extract shows strong antifungal activity against C. albicans. S. podophyllum could be applied in the fields of medical and pharmaceuticals for formulation of new drugs. Abbreviation used: AgNO3: Silver nitrate, MIC: Minimum inhibitory concentration, MTCC: Microbial type culture collection, SPR: Surface plasmon resonance, UV: Ultraviolet, XRD: X-ray diffraction
  - 2,579 164
A new highly selective and specific anti-puerarin polyclonal antibody for determination of puerarin using a mannich reaction hapten conjugate
Orapin Udomsin, Supaluk Krittanai, Tharita Kitisripanya, Hiroyuki Tanaka, Waraporn Putalun
October-December 2017, 13(52):845-851
DOI:10.4103/0973-1296.224331  PMID:29491643
Background: Puerarin (PUE) is a phytoestrogen found in Pueraria candollei and Pueraria lobata. These plants are substantial for traditional medicine in various Asian countries. PUE is a key marker that can be found only in the Pueraria species. Objective: To establish the method for determination of PUE content which is required for quality control of pharmaceutical products. Materials and Methods: PUE-cationized bovine serum albumin conjugate was created via Mannich reaction. After the rabbit immunization, the obtain anti-PUE polyclonal antibody (PAb) was used to develop an enzyme-linked immunosorbent assay (ELISA). Results: An anti-PUE PAb possess a great sensitivity and specificity. The cross-reactivity analysis shows no cross-reaction of an established antibody against other substances. In addition, we successfully developed an indirect competitive ELISA (icELISA) for the quantitative analysis of PUE. The result of method validation conforms to acceptance criteria and correlates with high-performance liquid chromatography, the reference method. The icELISA was applied to determine PUE content in Pueraria spp. plant samples and its derived pharmaceutical products. Conclusion: This highly specific immunogen was created from the Mannich reaction. An icELISA can also be applied to other research propose in the further studies. Abbreviations used: PUE: Puerarin; PAb: Polyclonal antibody; ELISA: Enzyme-linked immunosorbent assay; icELISA: Indirect competitive ELISA; cBSA: Cationized bovine serum albumin.
  - 1,338 83
Mechanistic In vitro Evaluation of Prosopis farcta roots potential as an antidiabetic folk medicinal plant
Saba Feyzmand, Behzad Shahbazi, Marzieh Marami, Gholamreza Bahrami, Ali Fattahi, Yalda Shokoohinia
October-December 2017, 13(52):852-859
DOI:10.4103/0973-1296.224332  PMID:29491644
Objective: Prosopis farcta has been used as a traditional herbal medicine for treating Diabetes mellitus. The aim of this study is to investigate the antidiabetic mechanisms of infusion (INF) extract of P. farcta and discovering the active extract for the first time. Materials and Methods: Six different extracts of P. farcta were prepared using five different solvents (ethanol, n-hexane, acetone, ethanol:water (1:1 v/v), and water). Cytotoxicity and cell proliferation assays were performed on mouse pancreatic β-cells (β-TC3) using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium method. The effects of P. farcta on glucose metabolism (in a hepatocellular carcinoma cell line [HepG2]) and glucose diffusion across a dialysis membrane (as a model of cellular glucose absorption) were evaluated. The protective effect of various P. farcta extracts on cytotoxicity, mitochondrial membrane potential (MMP), and streptozotocin (STZ)-induced apoptosis in β-TC3 cells was investigated. Results: Cytotoxicity study indicated that extracts were safe on β-TC3and HepG2 (≤0.5 mg/ml). INF protected β-TC3 cells from apoptosis induced by STZ and improved cell viability for 20% and significantly decrease depolarization of MMP (P < 0.005). The results showed that INF inhabited breaking/streaking the DNA. Proliferation study showed no significant increase in the number of cells either at single or multiple doses. In moderate hyperglycemia (11.1 mmol/l), a significant glucose-lowering effect was observed but glucose diffusion was not the probable mechanism of extracts antidiabetic effect. In conclusion, only INF, the traditionally used extract, has an antidiabetic potential by attenuating the death and apoptosis induced by STZ in β-TC3 cells and increase glucose consumption. Conclusion: The present study demonstrates that only INF extract have an antidiabetic potential by attenuating the death and apoptosis induced by STZ in β-TC3 cells and increase glucose consumption. Abbreviations used: AC: Acetone extract; ANOVA: Analysis of variance; BSA: Bovine serum albumin; β-TC3: Mouse pancreatic β-cells; DMEM: Dulbecco modified Eagle medium; DMSO: Dimethyl sulfoxide; ETH: Ethyl acetate extract; FBS: Fetal bovine serum; HDETH: Hydroethanolic extract; HepG2: Hepatocellular carcinoma cell line; HEX: Hexane extract; INF: Infusion; KUMS: Kermanshah University of Medical Sciences; MMP: Mitochondrial membrane potential; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium; NaCl: Natrium chloride; OD: Optical density; spp: Species; STZ: Streptozotocin; Tag: T-antigen; USA: United States of America.
  - 1,741 99
Hepatoprotective effects of nonpolar extracts from inflorescences of thistles Cirsium vulgare and Cirsium ehrenbergii on Acute liver damage in rat
Eduardo Fernández-Martínez, Maribel Jiménez-Santana, Mónica Centeno-Álvarez, Jose Martín Torres-Valencia, Mineko Shibayama, Raquel Cariño-Cortés
October-December 2017, 13(52):860-867
DOI:10.4103/0973-1296.224333  PMID:29491645
Background: Drugs for the treatment of liver diseases are scarce and not effective enough. Some species of the genus Cirsium possess hepatoprotective activity. There are no studies on the hepatoprotective effects of nonpolar extracts from inflorescences of thistles Cirsium vulgare and Cirsium ehrenbergii, and there are few reports on their chemical composition. Objective: The aim is to obtain the hexane extract from inflorescences of both thistles and to identify preliminarily their main chemical component, and to evaluate the hepatoprotective properties of the extracts. Materials and Methods: Hexane extracts were obtained using a Soxhlet apparatus. The chemical composition was analyzed using infrared spectra and gas chromatography-mass spectrometry. Two doses (250 and 500 mg/kg, p.o.) of both extracts were administered to assess their hepatoprotective effect on acute carbon tetrachloride (TC)-induced liver damage in rats using biochemical markers of necrosis, cholestasis, functionality, oxidative stress, and histological analysis. Results: Extracts were shown to have a very similar chemical profile. Their major constituent seems to be lupeol acetate. The two doses of both extracts demonstrated comparable hepatoprotective properties because they significantly diminished all the liver injury indicators (P < 0.05) and were corroborated using histopathology. Conclusion: This is the first study on the hepatoprotective effects of nonpolar extracts from inflorescences of thistles C. vulgare and C. ehrenbergii. Hexane extracts administration totally prevented the acute TC-induced liver damage. The preliminary chemical analysis strongly suggests the lupeol acetate as their major constituent. Lupeol and its derivatives have been previously reported as antiinflammatory and hepatoprotective agents. Abbreviations used: TC: Carbon tetrachloride; FT-IR: Fourier transform Infrared spectroscopy; GC-MS: Gas chromatography – Mass spectrometry; V: Vehicle; E: Extract; Ecv: Extract of Cirsium vulgare; Ece: Extract of Cirsium ehrenbergii; AP: Alkaline phosphatase; GGTP: γ-Glutamyl transpeptidase; ALT: Alanine aminotransferase; DB: Direct bilirubin; TB: Total bilirubin; LP: Lipid peroxidation; MDA: Malondialdehyde; NO: Nitric oxide; TNF-α: Tumor necrosis factor-α.
  - 3,034 24
The natural compound dansameum reduces foam cell formation by downregulating CD36 and peroxisome proliferator-activated receptor-gamma; Expression
Kang-Seo Park, Sang Hyun Ahn, Kang Pa Lee, Sun-Young Park, Jin Hong Cheon, Jun-Yong Choi, Kibong Kim
October-December 2017, 13(52):868-874
DOI:10.4103/0973-1296.224334  PMID:29491646
Background: Atherosclerosis-induced vascular disorders are major causes of death in most western countries. During the development of atherosclerotic lesions, foam cell formation is essential and formed through the expression of CD36 and the peroxisome proliferator-activated receptor gamma (PPAR-γ). Objective: To investigate whether dansameum extract (DSE) could show anti-atherosclerotic effect through down-regulating cellular redox state including CD36 and PARP-γ expression in oxidative low-density lipoprotein (oxLDL)-treated RAW264.7 cells and on differentiated foam cells in ApoE Knockout (ApoE-/-) mice. Materials and Methods: The Korean polyherbal medicine DSE was prepared from three plants in the following proportions: 40 g of Salvia miltiorrhiza root, 4 g of Amomumxanthioides fruit, and 4 g of Santalum album lignum. The immunohistochemistry and reverse transcription-polymerase chain reaction was used for analysis of protein and mRNA involved in foam cell formation. Results: We first showed that effects of DSE on foam cell formation in both oxLDL-induced RAW264.7 cells and in blood vessels from apolipoprotein E deficientApoE-/- mice with high fat diet-fed. DSE treatment significantly reduced the expression of CD36 and PPAR-γ in oxLDL-stimulated RAW264.7 cells and ApoE-/-mice, in the latter case by regulating heme oxygenase-1. Furthermore, DSE treatment also reduced cellular lipid content in vitro and in vivo experiments. Conclusion: Our data suggest that DSE may have anti-atherosclerotic properties through regulating foam cell formation. Abbreviations used: DSE: Dansameum extract, PPAR-γ: Peroxisome proliferator-activated receptor γ, HO-1: Heme oxygenase-1, CVD: Cardiovascular diseases
  - 1,589 122
Antioxidant activity of the essential oil and its major terpenes of Satureja macrostema (Moc. and Sessé ex Benth.) Briq.
Rafael Torres-Martínez, Yolanda Magdalena García-Rodríguez, Patricia Ríos-Chávez, Alfredo Saavedra-Molina, Joel Edmundo López-Meza, Alejandra Ochoa-Zarzosa, Rafael Salgado Garciglia
October-December 2017, 13(52):875-880
DOI:10.4103/0973-1296.224335  PMID:29491647
Background: The aim of this study was to investigate the in vitro antioxidant activity of Satureja macrostema (Moc. and Sessé ex Benth.) Briq. (Lamiaceae) essential oil, a Mexican medicinal plant known as nurite. Materials and Methods: Fresh aerial parts of S. macrostema plants cultivated in greenhouse for 3 months were subjected to hydrodistillation in a Clevenger apparatus to obtain essential oil. Volatile compounds were identified by gas chromatography (GC) and GC/mass spectrometry. Antioxidant effectiveness of essential oil and its major terpenes of S. macrostema was examined by three different radical scavenging methods: 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and total antioxidant capacity (TAC). The concentrations tested were 0.001, 0.01, 0.1, and 1 mg/mL. Results: The major volatile compounds were caryophyllene, limonene, linalool, pulegone, menthone, and thymol. S. macrostema essential oil showed the highest free radical scavenging activity with DPPH and ABTS methods (53.10% and 92.12%, respectively) at 1 mg/mL and 98% with TAC method at 0.1 mg/mL. Thymol exerted the highest antioxidant capacity with 0.1 mg/mL, reaching 83.38%, 96.96%, and 98.57% by DPPH, ABTS, and TAC methods. Caryophyllene, limonene, linalool, pulegone, and menthone exhibited an antioxidant capacity <25% with the DPPH and ABTS methods; however, limonene showed a TAC of 85.41% with 0.01 mg/mL. Conclusion: The essential oil of S. macrostema and thymol showed a free radical scavenging activity close to that of the synthetic butylated hydroxytoluene. Abbreviations used: GC: Gas Chromatography; DPPH: 2,2-diphenyl-1-picrylhydrazyl; ABTS: 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid; TAC: Total antioxidant capacity.
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Simultaneous quantification of forskolin and Iso-forskolin in Coleus forskohlii (Wild.) Briq. and identification of elite chemotype, collected from eastern ghats (India)
Pushpendra Kumar Shukla, Ankita Misra, Manish Kumar, Jaichand , Kuldeep Singh, Juber Akhtar, Sharad Srivastava, Pawan K Agrawal, Ajay K Singh Rawat
October-December 2017, 13(52):881-885
DOI:10.4103/0973-1296.224336  PMID:29491648
Background: Coleus forskohlii is a well-known industrially important medicinal plant, for its high forskolin content. Objective: A simple, selective, and sensitive high-performance thin layer chromatography (HPTLC) method was developed and validated for simultaneous quantification of forskolin and iso-forskolin in C. forskohlii germplasm collected from the Eastern Ghats, India. Materials and Methods: Chromatographic separation of the targeted marker(s) was obtained on precoated silica plates using toluene: ethyl acetate: methanol (90:30:0.5, v/v/v) as the mobile phase. Results: Densitometric quantification of forskolin and iso-forskolin was carried out at 545 nm. Forskolin and iso-forskolin were identified by comparing the ultraviolet spectra of standard and sample track at Rfof 0.64 ± 0.02 and 0.36 ± 0.01, after derivatization with anisaldehyde sulfuric acid reagent. The linearity of both the analytes was obtained in the range of 300–1200 ng/spot with the regression coefficient (R2) of 0.991 and 0.986. Recovery of analyte (s) at three levels, namely, 100, 150, and 200 ng/spot was found to be 100.46% ± 0.29%, 99.64% ± 0.33%, 100.02% ± 0.76% and 99.76% ± 0.62%, 99.56% ± 0.35%, 100.02% ± 0.22%, respectively, for forskolin and iso-forskolin. The content of forskolin and iso-forskolin varies from 0.046% to 0.187% and 0.002% to 0.077%, respectively (dry weight basis), the maximum content of both the markers was found in NBC-31, from Thakurwada, Maharashtra. Conclusion: The developed HPTLC method was linear, accurate, and reliable as per the International Council for Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use guidelines. The study aids in the identification of elite chemotype for commercial prospection of industrially viable medicinal crop.
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Methyl jasmonate and salicylic acid enhanced the production of ursolic and oleanolic acid in callus cultures of Lepechinia Caulescens
Víctor M Vergara Martínez, Samuel E Estrada-Soto, José de Jesús Arellano-García, Julio C Rivera-Leyva, Patricia Castillo-España, Angélica Flores Flores, Alexandre T Cardoso-Taketa, Irene Perea-Arango
October-December 2017, 13(52):886-889
DOI:10.4103/pm.pm_77_17  PMID:29491649
Background: The production of triterpenes from plants for pharmacological purposes varies in concentration, due to genetic and environmental factors. In vitro culture enables the control and increase of these bioactive molecules. Objective: To evaluate the effect of plant growth regulators and elicitors in the induction of calli and the production of ursolic acid (UA) and oleanolic acid (OA) in Lepechinia caulescens. Materials and Methods: Leaf explants were exposed for the induction of calli at different concentrations and combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). Methyl jasmonate (MJ) and salicylic acid were used as elicitors. High-performance liquid chromatography method was used to quantify UA and OA content in each treatment. Results: Treatment with 3.0 mg/L of 2,4-D and 0.1 mg/L of BAP produced the best results for calli induction and production of UA (1.57 mg/g dry weight [DW]) and OA (1.13 mg/g DW). Both elicitors facilitated the accumulation of triterpenes. Conclusion: The combination of auxins and cytokinins showed favorable results for the induction of calli. Variation concerning the accumulation of UA and OA was observed between treatments. MJ increased the production of triterpenes five times after 8 h of exposure, compared to control treatment. There is a greater accumulation of UA (16.58 mg/g DW) and OA (1.94 mg/g DW) in leaves of wild plants. Abbreviations used: 2,4-D: 2,4-dichlorophenoxyacetic acid, BAP: 6-benzylaminopurine, DW: Dry weight, MJ: Methyl jasmonate, OA: Oleanolic acid, PGRs: Plant growth regulators, UA: Ursolic acid, SA: Salicylic acid.
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Liquid chromatography mass spectrometry analysis and cytotoxicity of Asparagus adscendens roots against human cancer cell lines
Kashif Maqbool Khan, Lutfun Nahar, Abdul Mannan, Muhammad Arfan, Ghazanfar Ali Khan, Afaf Al-Groshi, Andrew Evans, Nicola M Dempster, Fyaz M D. Ismail, Satyajit D Sarker
October-December 2017, 13(52):890-894
DOI:10.4103/pm.pm_136_17  PMID:29491650
Background: Asparagus adscendens Roxb. (Asparagaceae), is native to the Himalayas. This plant has been used in the prevention and effective treatment of various forms of cancers. Objective: This paper reports, for the first time, on the cytotoxicity of the methanol (MeOH) extract of the roots of A. adscendens and its solid-phase extraction (SPE) fractions against four human carcinoma cell lines and LC-ESI-QTOF-MS analysis of the SPE fractions. Materials and Methods: Finely powdered roots of A. adscendens were macerated in methanol and extracted through SPE using gradient solvent system (water: methanol) proceeded for analysis on LC-ESI-QTOF-MS and cytotoxicity against four human carcinoma cell lines: breast (MCF7), liver (HEPG2), lung (A549), and urinary bladder (EJ138), using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide assay. Results: The MeOH extract and four SPE fractions exhibited cytotoxicity against all cell lines with the IC50values ranging from 6 to 79 μg/mL. As observed in other Asparagus species, the presence of saponins and sapogenins in the SPE fractions was evident in the liquid chromatography-mass spectrometry data. Conclusion: It is reasonable to assume that the cytotoxicity of the MeOH extract of the roots of A. adscendens and its SPE fractions, at least partly, due to the presence of saponins and their aglycones. This suggests that A. adscendens could be exploited as a potential source of cytotoxic compounds with putative anticancer potential. Abbreviation used: SPE: Solid-phase extraction, MCF7: Breast cancer cell line, HEPG2: Liver cancer cell line, A549: Lung liver cancer cell line, EJ138: Urinary bladder cancer cell line, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide, LC-MS: Liquid chromatography-mass spectrometry.
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Studying the Inhibitory Effect of Quercetin and Thymoquinone on Human Cytochrome P450 Enzyme Activities
Fawzy Elbarbry, Aimy Ung, Khaled Abdelkawy
October-December 2017, 13(52):895-899
DOI:10.4103/0973-1296.224342  PMID:29491651
Background: Quercetin (QR) and thymoquinone (TQ) are herbal remedies that are currently extensively used by the general population to prevent and treat various chronic conditions. Therefore, investigating the potential of pharmacokinetic interactions caused by the concomitant use of these herbal remedies and conventional medicine is warranted to ensure patient safety. Purpose of the Study: This study was conducted to determine the inhibitory effect of QR and TQ, two commonly used remedies, on the activities of selected cytochrome P450 (CYP) enzymes that play an important role in drug metabolism and/or toxicology. Materials and Methods: The in vitro studies were conducted using fluorescence-based high throughput assays using human c-DNA baculovirus expressed CYP enzymes. For measuring CYP2E1 activity, a validated High-performance liquid chromatography (HPLC) assay was utilized to measure the formation of 6-hydroxychlorzoxazone. Results: The obtained half-maximum inhibitory concentration values with known positive control inhibitors of this study were comparable to the published values indicating accurate experimental techniques. Although QR did not show any significant effect on CYP1A2 and CYP2E1, it exhibited a strong inhibitory effect against CYP2D6 and a moderate effect against CYP2C19 and CYP3A4. On the other hand, TQ demonstrated a strong and a moderate inhibitory effect against CYP3A4 and CYP2C19, respectively. Conclusions: The findings of this study may indicate that consumption of QR or TQ, in the form of food or dietary supplements, with drugs that are metabolized by CYP2C19, CYP2D6, or CYP3A4 may cause significant herb-drug interactions. Abbreviations used: ABT: Aminobenztriazole, BZF: 7,8 Benzoflavone, CYP: Cytochrome P450, GB: Gingko Biloba, IC50: Half-maximum inhibitory concentration, KTZ: Ketoconazole, QND: Quinidine, QR: Quercetin, TCP: Tranylcypromine, TQ: Thymoquinone.
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In Vitro bioassay-guided isolation of radioprotective fractions from extracts of Pinus koraiensis bark
Keli Yun, Jian-Hai Bai, ZhenYu Wang
October-December 2017, 13(52):712-718
DOI:10.4103/pm.pm_409_16  PMID:29200738
Objective: The aim of this study was to evaluate radioprotective effect of extracts of Pinus koraiensis bark and its fractions on rat splenocytes by using bioassay-guided isolation in order to obtain the best active fraction. Materials and Methods: P. koraiensis bark was ground and extracted with water, 40% acetone, 95% ethanol. Bio-guided assay was selected as an evaluation method to further fractionate radioprotective component from P. koraiensis bark extract. Total phenolic and flavonoid contents in fractions were also measured. Rat splenocytes were prepared by using mechanical trituration method. DNA damage was assessed as comet parameters (tail DNA%, tail length, tail moment, olive tail moment). The levels of malondialdehyde (MDA), and activity of superoxide dismutase (SOD), catalase (CAT) in cultured rat splenocytes were also measured. Results: The radioprotective effects decreased from rutin >95% ethanol extracts of Pinus koraiensis bark (95EEP) >40AEP > WEP. The stimulating effects decreased from rutin > n-butanol extract (NBE) > EAE. The results demonstrate that there exists toxic ingredients (PEE and dichloromethane extract), proliferative-promoting, radioprotective component (EAE and NBE) in 95EEP. fraction eluted from n-butanol fractions of 95EEP with 50% methanol solution (NBEPKB-50ME), a fraction of NBE result from bio-guided isolation, demonstrates good radioprotective efficacy on rat splenocytes. NBEPKB-50ME pretreated rat splenocytes demonstrated progressively reduced levels of MDA when compared with γ-ray exposed cells. Different dose of NBEPKB-50ME pretreatment with 8 Gy-irration showed an increase in enzymatic antioxidant. Conclusions: Proliferative-promoting efficacy, radioprotective effect of different solvents extracts of the bark of P. koraiensis were investigated in this work. NBEPKB-50ME was the best elution in NBE, especially in restoring SOD, CAT activities, content of GSH, decreasing DNA damage. Abbreviations used: MDA: Malondialdehyde; SOD: Superoxide dismutase; CAT: Catalase; PEE: Petroleum ether Extract; DME: Dichloromethane extract; EAE: Ethyl acetate extract; NBE: n-butanol extract; WAP: Water extracts of Pinus koraiensis bark; 40AEP: 40% acetone extracts of Pinus koraiensis bark; 95EEP: 95% ethanol extracts of Pinus koraiensis bark; TPC: Total phenolic content; TFC: Total flavonoid content; NBEPKB-50ME: Fraction eluted from n-Butanol fractions of 95EEP with 50% methanol solution.
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