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   2017| January-March  | Volume 13 | Issue 49  
    Online since January 6, 2017

 
 
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ORIGINAL ARTICLES
Vincamine alleviates amyloid-β 25-35 peptides-induced cytotoxicity in PC12 cells
Jianfeng Han, Qiumin Qu, Jin Qiao, Jie Zhang
January-March 2017, 13(49):123-128
DOI:10.4103/0973-1296.196309  PMID:28216895
Objective: Vincamine is a plant alkaloid used clinically as a peripheral vasodilator that increases cerebral blood flow and oxygen and glucose utilization by neural tissue to combat the effect of aging. The main purpose of the present study is to investigate the influence of vincamine on amyloid-β 25-35 (Aβ25-35) induced cytotoxicityto gain a better understanding of the neuroprotective effects of this clinically used anti-Alzheimer's disease drug. Materials and Methods: Oxidative stress was assessed by measuring malondialdehydeglutathioneand superoxide dismutase (SOD) levels. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell apoptosis detection was performed using an Annexin-V-FITC Apoptosis Detection Kit. The production of reactive oxygen species (ROS) was determined using an ROS Assay Kit. Western blot detection was carried out to detect the protein expression. Results: Our studies showed that pretreatment with vincamine could reduce Aβ25-35 induced oxidative stress. Vincamine markedly inhibited cell apoptosis dose-dependently. More importantlyvincamine increased the phosphatidylinositol-3 kinase (PI3K)/Akt and Bcl-2 family protein ratios on preincubation with cells for 2 h. Conclusion: Above observation led us to assume that one possible mechanism of vincamine protects Aβ25-35-induced cell death could be through upregulation of SOD and activation of the PI3K/Akt pathway. Abbreviation used: Aβ25-35: Amyloid-β 25-35; AD: Alzheimer's disease; BCA: Bicinchoninic acid; GSH: glutathione; PBS: Phosphate buffered solution; SDS: Sodium dodecylsulphate; SOD: Superoxide dismutase
  5,401 282 4
Anti-plasmodial and chloroquine resistance suppressive effects of embelin
OI Zaid, R Abd Majid, MS Hasidah, MN Sabariah, K Al-Zihiry, Sattar Rahi, R Basir
January-March 2017, 13(49):48-55
DOI:10.4103/0973-1296.203982  PMID:28479726
Background: Emergence of chloroquine (CQ) resistance among different strains of Plasmodium falciparum is the worst catastrophe that has ever perplexed the dedicated efforts to eradicate malaria. This urged the scientists to search for new alternatives or sensitizers to augment its antiplasmodium effect. Materials and Method: In this experiment, the potential of embelin, isolated from Embelia ribes, to inhibit the growth and sensitize CQ action was screened using SYBRE-green-I based drug sensitivity and isobologram assays, respectively. Its effect on red blood cells stability was screened to assess its safety. To explore its molecular mechanism, its effect on plasmodial Hemozoin and the in vitro β-hematin formation was screened as well. Furthermore, its anti-oxidant activity was measured using the conventional in vitro tests and its molecular characters were obtained using Molispiration program. Results: The results showed that its anti-plasmodial effect was weaker than CQ but synergism was obtained when they were combined at ratios lower than 5:5 CQ/embelin. Furthermore, β-hematin formation was inhibited by embelin without showing any synergism after mixing with CQ. Conclusion: Overall, embelin is not ideal to be suggested as a conventional antiplasmodium but it has a potential to ameliorate CQ resistance. Furthermore, its action is not related to its impact on hemozoin formation. Further, investigations are recommended to illustrate its detailed mechanism of action. Abbreviation used: CQ-DV-PBS-HEPES: Chloroquine-Digestive vacuole-Phosphate-buffer-saline-4-(2-hydroxyethyl-1-piperazin-ethan-sulphoni-acid), EDTA: Ethylen-diamin-tetra-acetic-acid, g.m.wt: Gram molecular weight, cMCM: Complete-malaria-culture-medium, Hct: Hematocrite, PRBCs: Parasitized-redblood-cells, nRBCs: Normal-red-blood-cells, RT: Room temperature, IC: Inhibitory concentration, FIC: Fractional inhibitory concentration, iCM: Incomplete-culturemedium, BSA: Bovin serum albumin, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, DPPH: 2,2-diphenyl-1- picrylhydrazy, BHT: Butylatedhydroxyl-toleuen, PSA: Polar surface area, ClogP: Log partition coefficient (octanol/water), GPCR: G-protein-coupled-receptors, DMSO: Dimethylsulphoxide, NaOH: Sodium hydroxide
  5,328 134 1
Medicinal plants used by traditional healers in Jordan, the Tafila region
Abeer Abdelhalim, Talal Aburjai, Jane Hanrahan, Heba Abdel-Halim
January-March 2017, 13(49):95-101
DOI:10.4103/0973-1296.203975  PMID:28479733
Background: The reporting of the medicinal plants and their traditional uses is important in order to prevent this knowledge from being lost. The aims of this study were to collect information concerning the traditional use of medicinal plants in the region of Tafila; identify the most important medicinal plants; determine the relative importance of the species surveyed; and calculate the informant consensus factor (Fic) in relation to medicinal plant use. Materials and Methods: Data on the traditional medicinal uses of local plants were collected using qualitative tools. The informant consensus factor (Fic) for the category of aliments and the use value (UV) of the plant species were calculated. Results and Conclusions: The survey revealed that 41 plant species are still in use in Tafila for the treatments of various diseases. Problems of the digestive system had the highest Fic values, while Allium cepa L. and Matricaria aurea (Loefl.) Sch. Bip. scored the highest UV. Abbreviations used: Fic: Informants consensus factor, nur: number of use reports per each category, nt: number of taxa used, UV: use value of a species, U: number of uses per species, n: number of informants
  4,612 232 16
Effect of shankhpushpi on alcohol addiction in mice
Mahi Heba, Sana Faraz, Sugato Banerjee
January-March 2017, 13(49):148-153
DOI:10.4103/0973-1296.203976  PMID:28479740
Alcohol addiction is a worldwide problem. It has mainly two components: dependence and withdrawal. Characteristic properties of most anti-addictive compounds include anti-anxiety, anticonvulsant, antidepressant, and nootropic actions. Shankhpushpi (Convolvulus pluricaulis. Convolvulaceae), known ethnopharmacologically as brain tonic, possess all the properties mentioned above. Here, we screen shankhpushpi for possible anti-addictive potential. Effect of shankhpushpi churna was measured on ethanol withdrawal anxiety using elevated plus maze. The role of shankhpushpi on chronic ethanol consumption (21 days) was measured using two bottle choice protocol of voluntary drinking. We also measured the effect of the above herb on cortico-hippocampal GABA levels. Shankhpushpi was found to reduce alcohol withdrawal anxiety in a dose-dependent manner. The herb also decreased ethanol intake and increased water intake significantly (P < 0.001) after 4 days of administration. Both these effects were blocked (P < 0.001) by GABAA antagonist suggesting the role of GABAA receptor. Chronic administration of shankhpushpi also significantly (P < 0.01) increased cortico-hippocampal GABA levels in mice. Shankhpushpi reduced both alcohol dependence and withdrawal in a GABAA-dependent manner, thus showing anti-addictive potential. Abbreviations used: GABA: Gamma-Aminobutyric Acid, HIV: Human Immunodeficiency Virus, CNS: Central Nervous System
  4,209 170 7
Preparation of ginseng extract with enhanced levels of ginsenosides Rg1 and Rb1 using high hydrostatic pressure and polysaccharide hydrolases
Sasikumar Arunachalam Palaniyandi, Joo-Won Suh, Seung Hwan Yang
January-March 2017, 13(49):142-147
DOI:10.4103/0973-1296.203992  PMID:28479739
Background: Ginsenosides are the principal components responsible for the pharmacological activities of ginseng. Ginsenosides Rg1 and Rb1 are the major compounds recognized as marker substances for quality control of ginseng-based products. These major compounds can be transformed to several pharmacologically active minor ginsenosides by chemical, microbial, and enzymatic means. Materials and Methods: In the present study, a combination of polysaccharide hydrolases and high hydrostatic pressure (HHP) were used to extract ginseng saponins enriched with ginsenosides Rg1 and Rb1. Temperature, pH, time, ginseng-to-water ratio, and pressure were optimized to obtain the maximum amount of Rg1 and Rb1 in the resulting extract using commercial polysaccharide hydrolases. Results: This study showed that treatment with a combination of cellulase, amylase, and pectinase at 100 MPa pressure, pH 4.8, and 45°C for 12 h resulted in higher Rg1 and Rb1 levels in the extract. Conclusion: This study describes a cheap and ecofriendly method for preparing ginseng extract enriched with Rg1 and Rb1. Abbreviations used: ATCC: American Type Culture Collection, Mpa: Mega Pascal
  4,055 293 3
Effect of a novel ashwagandha-based herbomineral formulation on pro-inflammatory cytokines expression in mouse splenocyte cells: A potential immunomodulator
Mahendra Kumar Trivedi, Sambhu Charan Mondal, Mayank Gangwar, Snehasis Jana
January-March 2017, 13(49):90-94
DOI:10.4103/0973-1296.197709  PMID:28479732
Background: Herbomineral formulations are momentous in an audience of worldwide by virtue of their holistic approach to life. These formulations are widely used as complementary therapies in immunocompromised patients including cancer. Still, there is the need of cost-effective and safe herbomineral-based formulation that can modulate immune response by the regulation of cytokines cascades. Objective: Current study, we investigated immunomodulatory effect of TEBEH in LPS-induced cytokines expression levels in mouse splenocytes in vitro. Materials and Methods: The most effective and safe concentrations of TEBEH were chosen by determining the cell viability of splenocytes using MTT assay. The pro-inflammatory cytokines such as TNF-α, IL-1β, MIP-1α, and IFN-γ were measured in cell supernatants using ELISA. Results: MTT data showed TEBEH formulation was found safe up to 10.53 µg/mL. At noncytotoxic concentrations (0.00001053–10.53 µg/mL), TEBEH significantly (P ≤ 0.001) inhibited the expressions of TNF-α, IL-1β, and MIP-1α in mouse splenocytes as compared with vehicle control. Conclusion: In summary, TEBEH may indeed promote an anti-inflammatory environment by suppression of pro-inflammatory cytokines. These observations indicated that TEBEH has potential effects in downregulating the immune system and might be developed as a useful anti-inflammatory product for various inflammatory disorders. SUMMARYThe present study was undertaken to evaluate an immunomodulatory effect of the herbomineral formulation in LPS-induced mouse splenocytes with the measurement of cytokines expression such as TNF-α, IL-1β, MIP-1α and IFN-γ. The results showed that the expression of TNF-α, IL-1β, and MIP-1α was significantly down-regulated while, IFN-γ was significantly up-regulated in mouse splenocytes. It is hypothesized that modulation of the proinflammatory cytokines might occur via NF-kB pathway. Therefore, the herbomineral test formulation might act as an effective anti-inflammatory and immunomodulatory product, and this can be used as a complementary and alternative treatment for the prevention of various types of inflammatory and auto-immune disorders. Abbreviations used: LPS: Lipopolysaccharide, IL: Interleukin; NF-kB: Nuclear factor kappa-B, TNF-α: Tumor necrosis factor alpha, MIP-1α: Macrophage inflammatory protein-1α, IFN-γ: Interferon, MTT: 3-(4, 5-diamethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium), ELISa: Enzyme linked immune sorbent assay, ANOVa: analysis of variance,
  3,948 266 6
Potential lipid-lowering effects of Eleusine indica (L) Gaertn. Extract on high-fat-diet-induced hyperlipidemic rats
Siew Ling Ong, Koteswara Rao Nalamolu, How Yee Lai
January-March 2017, 13(49):1-9
DOI:10.4103/0973-1296.203986  PMID:28479718
Background: To date, anti-obesity agents based on natural products are tested for their potential using lipase inhibition assay through the interference of hydrolysis of fat by lipase resulting in reduced fat absorption without altering the central mechanisms. Previous screening study had indicated strong anti-obesity potential in Eleusine indica (E. indica), but to date, no pharmacologic studies have been reported so far. Objective: This study was performed to investigate the lipid-lowering effects of E. indica using both in vitro and in vivo models. Methods: The crude methanolic extract of E. indica was fractionated using hexane (H-Ei), dichloromethane (DCM-Ei), ethyl acetate (EA-Ei), butanol (B-Ei), and water (W-Ei). All the extracts were tested for antilipase activity using porcine pancreatic lipase. Because H-Ei showed the highest inhibition, it was further subjected to chemical profiling using high-performance liquid chromatography. Subsequently, oral toxicity analysis of H-Ei was performed [Organization for Economic Cooperation and Development guidelines using fixed dose procedure (No. 420)]; efficacy analysis was performed using high-fat diet (HFD)-induced hyperlipidemic female Sprague–Dawley rats. Results: According to the toxicity and efficacy analyses, H-Ei did not demonstrate any noticeable biochemical toxicity or physiologic abnormalities and did not cause any tissue damage as per histologic analysis. Furthermore, H-Ei significantly reduced body weight and improved serum profile and did not show hepatotoxicity and nephrotoxicity based on the serum profile. Moreover, H-Ei alleviated HFD-induced hepatosteatosis and ameliorated induced adiposity in both visceral and subcutaneous adipose tissue. Conclusion: Our results demonstrate that H-Ei effectively improved hyperlipidemia. Further studies to explore its possibility as an alternative pharmacologic agent to treat obesity are warranted. Abbreviation Used: ALT: Alanine transaminase; AST: Aspartate transaminase; B-Ei: Butanol extract of E. indica; DCM-Ei: Dichloromethane extract of E. indica; EA-Ei: Ethyl acetate extract of E. indica; GHS: Globally Harmonized System; HDL: High-density lipoprotein; H-Ei: Hexane extract of E. indica; HFD: High-fat diet; HPLC: High-performance liquid chromatography; LDL: Low-density lipoprotein; NFD: Normal fed diet; PPL: Porcine pancreatic lipase; SEM: Standard error of mean; SD: Standard deviation; TC: Total cholesterol; TG: Triglycerides; W-Ei: Water extract of E. indica.
  3,935 264 2
Caffeic acid phenethyl ester (CAPE) prevents development of STZ-ICV induced dementia in rats
Manish Kumar, Devinder Kaur, Nitin Bansal
January-March 2017, 13(49):10-15
DOI:10.4103/0973-1296.203974  PMID:28479719
Background: Chronic oxidative stress and inflammation severely affect the normal physiology of neurons and lead to neurodegenerative disorders such as Alzheimer's disease (AD). Polyphenols proved a boon in the prevention of dementia due to their antioxidant and neuroprotective potential. Caffeic acid phenethyl ester (CAPE) is a natural polyphenolic compound attributed with antioxidant, immunomodulatory, and neuroprotective properties. Objective: The present study investigates the effect of CAPE on experimental dementia in rats. Methods: Intracerebroventricle (ICV) injection of streptozotocin (STZ; 3 mg/kg) was given to Wistar rats (200 g, either sex) on days 1 and 3 to induce dementia of AD type. CAPE (3 and 6 mg/kg, i.p.) was administered to separate groups of rats for 28 successive days daily. Morris water maze and elevated plus maze served as exteroceptive behavioral models to measure the memory of the rats. Results: The present study illustrated that CAPE treatment for 28 consecutive days arrested the development of cognitive deficits in STZ-ICV-treated rats, that is, a significant (P < 0.05) reduction in the mean escape latency during acquisition trial and increased (P < 0.05) time spent in target quadrant during retrieval trial in Morris water maze test and reduction (P < 0.05) in transfer latency in elevated plus maze test. Furthermore, both the doses of CAPE when administered to rats that were previously treated with STZ-ICV prevented the rise of brain thiobarbituric acid reactive substance as well as TNF-α and simultaneously enhanced the GSH content. Conclusion: CAPE administration ameliorated STZ-ICV-induced dementia through the attenuation of oxidative stress and inflammation. Abbreviation used: AD: Alzheimer's disease, ANOVA: Analysis of Variance, aCSF: Artificial cerebrospinal fluid, CAPE: Caffeic acid phenethylester, EPM: Elevated plus maze, ELT: Escape latency time, GSH: Reduced glutathione, IL: Interleukin, ICV: Intracerebroventricular, MDA: Malondialdehyde, MEL: Mean escape latency, MWM: Morris water maze, NFTs: Neurofibrillary tangles, RNS: Reactive nitrogen species, ROS: Reactive oxygen species, SEM: Standard error of mean, STZ: Streptozotocin, TBARS: Thiobarbituric reactive substances, TSTQ: Time spent in target quadrant, TL: Transfer latency, TNF-α: Tumor necrosis factor alpha.
  3,894 277 12
Influence of capsaicin on inflammatory cytokines induced by lipopolysaccharide in myoblast cells under in vitro environment
Ke Shang, Touseef Amna, Musarat Amina, Nawal M Al-Musayeib, Salem S Al-Deyab, Inho Hwang
January-March 2017, 13(49):26-32
DOI:10.4103/0973-1296.203984  PMID:28479722
Background: Cellular damage initiated by reactive oxygen species (ROS) is the main cause of numerous severe diseases and therefore for this reason, the natural antioxidants have note worthy significance in human health. Capsaicin possesses noteworthy analgesic and anti-inflammatory properties. It also possesses healing effects for treatment of arthritis, diabetic neuropathy, gastric lesions, and cardiac excitability that is why it is incorporated in creams and gels. Objective: The present study was carried out to estimate the in vitro antioxidant and ROS scavenging activities of capsaicin against muscle precursor cells. Till date,no investigation has been carried out to study the effect of capsaicin on myoblasts.Materials and methods: Herein, the cytotoxicity was induced by endotoxin lipopolysaccharide (LPS) to analyze the effect of capsaicin on LPS induced inflammation and apoptosis on muscle cells. To find out the toxicity of endotoxin, myoblasts were exposed to different concentrations of LPS, viability and morphology was checkedby the means of CCK-8 test and microscopy, respectively. Apoptotic cell death was examined by fluorescence staining. Additionally, LPS-induced apoptosis was determined by mRNAexpression of calpain, caspase-3 and tumor necrosisfactor alpha (TNF-α), and were quantified by qRT-PCR. Results: The outcome of the presentstudy demonstrated that LPS stimulation generatestoxicity in dose-dependent manner. Pre-treatmentof myoblasts with capsaicin can considerably alleviate LPS-induced inflammation. Conclusion: In conclusion, this study indicates that dietetic supplementation of capsicum may help to alleviate/reduce the inflammatory effects and is therefore potent source of natural antioxidant agent which can be utilized to control muscle related diseases, such as myotube atrophy. Abbreviation used: AMP: Adenosine monophosphate, AO/EB: Acridine orange / Ethidium bromide, ATL: T-cell leukemi, CAP: Capsaicin, CCK-8: Cell counting Kit-8, CLSM: Laser Scanning Microscopy, DCF-DA: 2', 7'-dichlorofluorescein diacetate, DMEM: Dulbecco's modified Eagle's medium, DPPH: α, α-diphenyl-β-picrylhydrazyl, FBS: Fetal bovine serum, KA: Kainic acid, LPS: Lipopolysaccharide, MDA: Malondialdehyde, NF-kB: Nuclear factor kgene binding, PBS: Phosphate buffer saline, pNA: p-nitroanilide, RNW: RNase free water, ROS: Reactive oxygen species, TNF-α: Tumor necrosis factor alpha, TRPV1: Transient receptor potential vanilloid 1
  3,605 273 5
Effect of naringenin (A naturally occurring flavanone) against pilocarpine-induced status epilepticus and oxidative stress in mice
Sheeba Shakeel, Muneeb U Rehman, Nahida Tabassum, Umar Amin, Manzoor ur Rahman Mir
January-March 2017, 13(49):154-160
DOI:10.4103/0973-1296.203977  PMID:28479741
Background: Epilepsy is a disorder of the central nervous system characterized by recurrent seizures. It is a very common disease in which approximately 30% of patients do not respond favourably to treatment with anticonvulsants. Oxidative stress is associated with neuronal damage arising from epileptic seizures. The present study investigated the effects of naringenin in pilocarpine-induced epilepsy in mice. Naringenin, one of the most frequently occurring flavanone in citrus fruits, was evaluated for its shielding effect against the pilocarpine induced behavioural, oxidative and histopathological alterations in rodent model of epilepsy. Methodology: Epilepsy was induced by giving pilocarpine (300mg/kg) and sodium valproate (300mg/kg) was given as standard anti-epileptic drug Pilocarpine was administered (300 mg /kg body weight) intraperitoneally to the mice on 15th day while naringenin was administered orally (20 and 40 mg/kg body weight) for 15 days prior to administration of pilocarpine. Results: The intraperitoneal administration of pilocarpine enhanced lipid peroxidation, caused reduction in antioxidant enzymes, viz., catalase, superoxide dismutase and glutathione reductase. Treatment of mice orally with naringenin (20 mg/kg body weight and 40 mg/kg body weight) resulted in a significant decrease in lipid peroxidation. There was significant recovery of glutathione content and all the antioxidant enzymes studied. Also in case of behavioural parameters studied, naringenin showed decrease in seizure severity. All these changes were supported by histological observations, which revealed excellent improvement in neuronal damage. Conclusion: The higher dose of naringenin was more potent in our study and was comparable to the standard drug (sodium valproate) in effectiveness. Abbreviations used:6-OHDA: 6-hydroxydopamine, AED: Anti epileptic drugs, AIDS: Acquired immune deficiency syndrome, ANOVA: Analysis of variance, ATP: Adenosine triphosphate, CA: Cornu ammonis, CAT: Catalase, DG: Dentate gyrus, EDTA: Ethylenediamine tetra acetic acid, GR: Glutathione reductase, GSH: Glutathione reduced, HCl: Hydrochloric acid, IL-1β: Interleukin 1 beta, LPO: Lipid peroxidation, MDA: Malondialdehyde, NADPH: Nicotinamide adenine dinucleotide phosphate, PMS: post mitochondrial supernatant, SE: Status epilepticus, SEM: Standard error of the mean, SOD Superoxide dismutase, TBA: Thiobarbituric acid, TBARS: Thiobarbituric acid reactive substance, TLE: Temporal lobe epilepsy, TNF-α: Tumor necrosis factor alpha
  3,576 247 18
Depolarizing effects of daikenchuto on interstitial cells of cajal from mouse small intestine
Hyungwoo Kim, Hyun Jung Kim, Dongki Yang, Myeong Ho Jung, Byung Joo Kim
January-March 2017, 13(49):141-147
DOI:10.4103/0973-1296.196312  PMID:28216898
Background: Daikenchuto (DKT; TJ-100, TU-100)a traditional herbal medicineis used in modern medicine to treat gastrointestinal (GI) functional disorders. Interstitial cells of Cajal (ICCs) are the pacemaker cells of the GI tract and play important roles in the regulation of GI motility. Objective: The objective of this study was to investigate the effects of DKT on the pacemaker potentials (PPs) of cultured ICCs from murine small intestine. Materials and Methods: Enzymatic digestions were used to dissociate ICCs from mouse small intestine tissues. All experiments on ICCs were performed after 12 h of culture. The whole-cell patch-clamp configuration was used to record ICC PPs (current clamp mode). All experiments were performed at 30-32°C. Results: In current-clamp modeDKT depolarized and concentration-dependently decreased the amplitudes of PPs. Y25130 (a 5-HT3 receptor antagonist) or SB269970 (a 5-HT7 receptor antagonist) did not block DKT-induced PP depolarizationbut RS39604 (a 5-HT4 receptor antagonist) did. Methoctramine (a muscarinic M2 receptor antagonist) failed to block DKT-induced PP depolarizationbut pretreating 4-diphenylacetoxy-N-methylpiperidine methiodide (a muscarinic M3 receptor antagonist) facilitated blockade of DKT-induced PP depolarization. Pretreatment with an external Ca2+-free solution or thapsigargin abolished PPsand under these conditionsDKT did not induce PP depolarization. FurthermoreGinseng radix and Zingiberis rhizomes depolarized PPswhereas Zanthoxyli fructus fruit (the third component of DKT) hyperpolarized PPs. Conclusion: These results suggest that DKT depolarizes ICC PPs in an internal or external Ca2+-dependent manner by stimulating 5-HT4 and M3 receptors. Furthermorethe authors suspect that the component in DKT largely responsible for depolarization is probably also a component of Ginseng radix and Zingiberis rhizomes. Abbreviation used: DKT: DaikenchutoGI: GastrointestinalICCs: Interstitial cells of CajalPPs: Pacemaker Potentials.
  3,622 193 1
Flavonoids from whole plant of Euphorbia hirta and their evaluation against experimentally induced gastroesophageal reflux disease in rats
Shyam Sundar Gupta, Lubna Azmi, PK Mohapatra, Ch V Rao
January-March 2017, 13(49):127-134
DOI:10.4103/0973-1296.203987  PMID:28479737
Background:Euphorbia hirta possesses antibacterial, anti-inflammatory, galactogenic, antidiarrheal, antioxidant, hypoglycemic, antiasthmatic, antiamebic, antifungal, and antimalarial activities. Objective:The overall objective of the current study was the investigation of the whole plant extract of E. hirta and flavonoids from E. hirta on gastroesophageal reflux disease (GERD) in rats. Materials and Methods:The whole plant extract of E. hirta was characterized by analysis of flavonoids (HPLC: HPLC, UV, IR, MS and 1HNMR). GERD model was induced surgically in Wistar rats under pentobarbitone sodium anesthesia (50 mg/kg, i.p.) and the tissue esophagus and stomach were removed. The tissues were washed with physiological saline and were examined for GERD. The whole plant extract of E. hirta in doses of 50, 100, and 200 mg/kg were administered orally twice daily at 10:00 and 16:00 hours, respectively, for 5 days and kaempferol (100 mg/kg) or omeprazole (OMZ) in the dose of 30 mg/kg 1 hour prior to the induction of GERD. Control groups received suspension of 1% carboxymethyl cellulose in distilled water (10 mL/kg). Results: The levels of gastric wall mucus increased and of plasma histamine and H+, K+ ATPase significantly decreased in groups treated by both the plant extract and flavonoids. Both the plant extract and flavonoids reduced the lipid peroxidation and superoxide dismutase and increased the levels of catalase and reduced glutathione. Conclusions: The whole plant extract of E. hirta is attributed to its antisecretory, gastroprotective, and antioxidant potential as that of quercetin, rutin, kaempferol, and proton pump blocker (omeprazole) to treat GERD. Abbreviation used: 1HNMR: Proton Nuclear Magnetic Resonance Spectroscopy, CAT: Catalase, EHAE: Aqueous extract of Euphorbia hirta, EHEF: Ethyl Acetate Fractions of Euphorbia hirta, GERD: Gastroesophageal reflux disease, GSH: Reduced Glutathione, HPLC: High performance liquid chromatography, IR: Infrared spectroscopy, LPO: Lipid Peroxidase, MDA: Malondialdehyde, MS: Mass Spectroscopy, OMZ: Omeprazole, ROS: Reactive Oxygen Species, SOD: Superoxide dismutase, TBHQ: tert-Butylhydroquinone, TLC: Thin Layer Chromatography, UV: Ultraviolet, UV: Ultraviolet–Visible Spectroscopy
  3,571 236 6
In vitro anti-proliferative effect of Tephrosia purpurea on human hepatocellular carcinoma cells
Ramamoorthy Padmapriya, Loganathan Gayathri, Larance Ronsard, Mohammad A Akbarsha, Ramasamy Raveendran
January-March 2017, 13(49):16-21
DOI:10.4103/0973-1296.203981  PMID:28479720
Background: Tephrosia purpurea is an Indian herb used in traditional medicine to treat various diseases such as jaundice, asthma, liver and urinary disorders. However, the anti-cancer potential of T. purpurea on hepatocellular carcinoma (HCC) is poorly understood. Therefore, this study aims to investigate the anti-cancer activity of T. purpurea in HepG2 hepatocellular carcinoma cells. Methods: The leaves and root of T. purpurea were extracted with methanol using soxhlet apparatus. The cytotoxicity of the T. purpurea extracts in HepG2 cells was evaluated using MTT assay whereas the mode of cell death was examined by AOEB, Hoechst and JC1 staining under a fluorescence microscope. T. purpurea extracts-induced caspase-3 expression was investigated using colorimetric assay. Results: The leaves and root extracts inhibited HepG2 cell growth at the IC50 of 102.33 ± 10.26 µg/mL and 276.67 ± 20.43 µg/mL respectively at 24 h. Chromatin condensation, nuclear fragmentation, apoptotic bodies formation and mitochondrial membrane depolarization were observed in HepG2 cells treated with both extracts. The caspase-3 expression was significantly (p < 0.05) increased in extracts treated cells when compared to control. Conclusion: The leaves and root extracts of T. purpurea induce apoptosis mediated cell death in HepG2 cells. Abbreviation used: AO: acridine orange, DMSO: dimethyl sulfoxide, EB: ethidium bromide, IC50: the concentration at which 50% of cancer cells are dead, JC-1: 5, 5', 6, 6'-tetrachloro-1, 1', 3, 3'-tetraethyl-imidacarbocyanine iodide, MTT: 3-4, 5-dimethylthiazole-2-yl, 2,5-diphenyl tetrazolium bromide, PBS: phosphate-buffered saline, ΔΨm: mitochondrial trans-membrane potential
  3,497 200 1
Antidermatophytic and protease-inhibiting activities of zerumbone: A natural sesquiterpene from the rhizome of Zingiber zerumbet (L.) Roscoe ex J.E; Smith
Madhavankutty Jyothilakshmi, Mathew Jyothis, Gokulanathan Nair Hari Narayanan, Mukalel Sankunni Latha
January-March 2017, 13(49):2-6
DOI:10.4103/0973-1296.197649  PMID:28216875
Context: Due to increase in the number of patients with impaired immunity, incidence of dermatophytoses has increased considerably. Antidermatophytic agents with anti-inflammatory and protease-inhibiting activities will help in restricting inflammatory response associated with dermatophytoses. Aims: The present study aims to evaluate antidermatophytic and protease-inhibiting activities of zerumbone. Cytotoxicity was tested using Chang liver cell line as a preliminary step in toxicity study. Methods and Materials: Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of zerumbone purified from the rhizome of Zingiber zerumbet were determined against Epidermophyton floccosum var. nigricans, Microsporum canis, Microsporum gypseum, and Trichophyton rubrum. MIC was determined according to Clinical and Laboratory Standards Institute (CLSI) method M38-A2. Protease-inhibiting property was tested using trypsin as the enzyme. In vitro cytotoxic effect was studied using the MTT assay. Results: MIC of zerumbone was 8 mg/L against E. floccosum and M. canis and 16 mg/L for M. gypseum and T. rubrum. MFC of zerumbone was 64 mg/L against E. floccosum and M. canis and 128 mg/L for M. gypseum and T. rubrum. Zerumbone exhibited remarkable protease-inhibiting activity. In the MTT assay, IC50 values were 150 and 0.31 µg, respectively, for zerumbone and reference drug. Statistical Analysis Used: For protease inhibition, assay and cytotoxicity assay control and tests were done in triplicate and the results are expressed as mean ± SD, where n = 3. Conclusions: Zerumbone is a novel candidate for use in dermatophytoses therapy because of the combined antifungal, anti-inflammatory (unpublished results), and protease-inhibiting properties. Cytotoxicity of zerumbone was found to be very low compared with the reference drug. Abbreviations used: CFU: colony forming unit, CLSI: Clinical and Laboratory Standards Institute, COX: cyclooxygenase, DMSO: dimethyl sulphoxide, EDTA: ethylene diamine tetra acetic acid, FT-IR: Fourier transform–infra red spectroscopy, HPLC: high-performance liquid chromatography, LOX: lipoxygenase, IMTECH: Institute of Microbial Technology, LCMS: liquid chromatography mass spectrometry, MTT: 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, MTCC: microbial type culture collection, MFC: minimum fungicidal concentration, MIC: minimum inhibitory concentration, MPO: myeloperoxidase, NMR: nuclear magnetic resonance spectroscopy, PAR: proteinase-activated receptor, PBS: phosphate-buffered saline, TCA: trichloro acetic acid
  3,272 127 -
Prenatal developmental toxicity study of glycosides-based standardized fenugreek seed extract in rats
Pallavi O Deshpande, Vishwaraman Mohan, Mukul p Pore, Shailesh Gumaste, Prasad A Thakurdesai
January-March 2017, 13(49):135-141
DOI:10.4103/0973-1296.203978  PMID:28479738
Context: Glycoside-based standardized fenugreek seed extract (SFSE-G) demonstrated promising efficacy in animal models of immune-inflammatory conditions. Aim: The present study was aimed at embryo-fetal development toxicity evaluation of SFSE-G in Wistar rats as per guideline No. 414 of the Organization for Economic Co-operation and Development (OECD). Material and Methods: Mated female rats were randomized into four groups of 30 each and received oral doses of either SFSE-G at 250, 500, and 1000 mg/kg or vehicle (water) during the period of gestation (postconception) from gestational day 5 (GD5, an implantation day) until 1 day before cesarean sections (GD19). Maternal food consumption, body weights, and clinical signs were monitored throughout gestation. Cesarean sections were performed on GD20 and fetal observations (gravid uterine weight, implantation sites, early and late resorptions, live and dead fetuses) were recorded. Live fetuses were weighed and examined for external, visceral, and skeletal variations and malformations. Results: None of the SFSE-G-treated groups showed maternal and embryo–fetal toxicity. Occasional and incidental skeletal and visceral malformations were observed and found to be spontaneous and unrelated to the treatment. Conclusion: Oral exposure of SFSE-G during the prenatal period did not show significant maternal and embryo-fetal toxicity up to a dose of 1000 mg/kg in rats. Therefore, the no-observed-adverse-effect level for SFSE-G for prenatal oral exposure was considered to be 1000 mg/kg. Abbreviations used: CPCSEA: Committee for the Purpose of Control and Supervision of Experiments on Animals; GD: Gestational day; GRAS: Generally recognized as safe; HED: Human equivalent dose; NOAEL: No-observed adverse effect levels; OECD: Organization for Economic Co-operation and Development; SFSE-G: glycoside-based standardized fenugreek seed extract
  3,136 183 -
Biosynthesis, characterization, and biological activities of iron nanoparticles using Sesamum indicum seeds extract
Farah Bano, Muhammad Baber, Amjad Ali, Ziaullah Shah, Syed Aun Muhammad
January-March 2017, 13(49):33-36
DOI:10.4103/0973-1296.203985  PMID:28479723
Background: Iron nanoparticles (FeNPs) have got many biomedical and health applications because of biocompatible and nontoxic nature to humans. Objective: To synthesize the FeNPs using natural sources. Materials and Methods: In this study, simple and economical procedure was adopted for FeNPs synthesis. Sesame seeds were processed to obtain seed extract as a biological material for FeNPs production. FeNPs were characterized by Fourier transform infrared spectroscopy, X-ray diffraction, and scanning electron microscopic. Results: The average diameter of these FeNPs was 99 nm. These nanoparticles showed significant anti-typhoid activity (30 mm zone of inhibition) as compared to ciprofloxacin (32 mm) as standard. Furthermore, in vitro alpha-amylase inhibitory assay also showed moderate antidiabetic activity with more than 50% inhibition. Conclusion: This study would be helpful in understanding of nanoparticles synthesis from natural sources and ultimately will be used as potential alternative therapeutic agents. Abbreviations used: FeNPs: Iron Nano Particles; SEM: Scanning Electron Microscopy; MIC: Minimum Inhibitory Concentration; S. indicum: Sesamum Indicum.
  3,151 164 4
Cytotoxic activities of naturally occurring oleanane-, ursane-, and lupane-type triterpenes on HepG2 and AGS cells
Heejung Yang, Hyun Woo Kim, Young Choong Kim, Sang Hyun Sung
January-March 2017, 13(49):118-122
DOI:10.4103/0973-1296.196308  PMID:28216894
Background: It is well known that the naturally occurring modified triterpenes in plants have a wide diversity of chemical structures and biological functions. The lupane-oleanane-and ursane-type triterpenes are the three major members of natural triterpenes with a wide range of biological properties. A systematic approach is necessary to review their structures and biological activities according to the backbones and the different substituents. Objective: Thirty lupane-(L1-7)oleanane-(O1-14)and ursane-type (U1-9) triterpenes with structural diversity were examined to evaluate their cytotoxic activities against two cancer cell lineshuman hepatocellular carcinoma (HepG2) and AGS cells. Materials and Methods: They were isolated from Hedera helixJuglans sinensisand Pulsatilla koreana using a series of column chromatography methods and were treated to evaluate their cytotoxic activities against HepG2 and AGS human gastric adenocarcinoma cell. Furthertwo triterpenes showing the most potent activities were subjected to the apoptotic screening assay using flow cytometry. Results: The polar groupssuch as an oxo group at C-1a free hydroxyl at C-2C-3or C-23and a carboxylic moiety at C-28as well as the type of backboneexplicitly increased the cytotoxic activity on two cancer cells. O5 and U5 showed significantly the potent cytotoxic activity in comparison to other glycosidic triterpenes. In annexin-V/propidium iodide (PI) staining assaythe percentage of late apoptosis (annexin-V+/PI+) 12 and 24 h after treatment with O5 and U5 at 25 μM increased from 14.5% to 93.1% and from 46.4% to 49.1%respectivelyin AGS cells. The cytotoxicity induced by O5 showed a significant difference compared to U5 for 12 and 24 h. Conclusion: In the studywe can suggest the potent moieties which influence their cytotoxic activities against two cancer cells. The polar groups at C-1C-2C-3C-23and C-28 and the linkage of sugar moieties influenced the different cytotoxic activities.
  3,093 166 4
In-vitro wound healing effect of 15-hydroxyprostaglandin dehydrogenase inhibitor from plant
Sandeep Karna
January-March 2017, 13(49):122-126
DOI:10.4103/0973-1296.203971  PMID:28479736
Background: Prostaglandins (PGs) have short existence in vivo because they are rapidly metabolized by NAD+-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) to 15-ketoprostaglandins. Inhibition of 15-PGDH causes elevated level of PGE2 in cellular system. It will be valuable for the therapeutic management of diseases requiring elevated PGE2 levels, like wound healing. Objective: Ninety-eight plant samples were screened for the discovery of potent 15-PGDH inhibitor. Among them, top five plant extracts as potent 15-PGDH inhibitor were chosen to determine PGE2 release from HaCaT (Keratinocyte cell line) cell line. Finally, top 15-PGDH inhibitor was selected to evaluate in vitro wound healing effect on HaCaT scratch model. Method: The inhibitory activity for 15-PGDH inhibitors was evaluated using fluorescence spectrophotometer by measuring the formation of NADH at 468 nm following excitation at 340 nm. Cell viability assay and PGE2 release was evaluated in HaCaT cell line after treatment of 15-PGDH inhibitors. Scratches were made using sterile 200 μL on HaCaT cell and wound-healing effect was evaluated after treatment of 15-PGDH inhibitor. Results: 15-PGDH inhibitors elevated PGE2 levels in concentration-dependent manner. Ethanol extract of Artocarpus heterophyllus (EEAH), the most potent 15-PGDH inhibitor (IC50 = 0.62 µg/mL) with least cytotoxicity (IC50 = 670 µg/ml), elevated both intracellular and extracellular PGE2 levels. EEAH facilitated in-vitro wound healing in a HaCaT (Keratinocyte cell line) scratch model. Conclusion: EEAH might apply to treat dermal wounds by elevating PGE2 levels via COX-1 induction and 15-PGDH inhibition. Abbreviations used: 15-PGDH: 15-hydroxyprostaglandin dehydrogenase, COX: Cyclooxygenase, DTT: Dithiothreitol, DMEM: Dulbecco's modified Eagle's media, EEAH: Ethanol extract of Artocarpus heterophyllus, MRP4: Multidrug resistance 4, PGs: Prostaglandins, PGT: Prostaglandin transporter, SDS: Sodium dodecylsulfate
  3,015 220 2
Simultaneous determination of multiple ginsenosides in Panax ginseng herbal medicines with one single reference standard
Chunwei Wu, Qingxiao Guan, Shumei Wang, Yueying Rong
January-March 2017, 13(49):84-89
DOI:10.4103/pm.pm_274_16  PMID:28479731
Background: Root of Panax ginseng C. A. Mey (Renseng in Chinese) is a famous Traditional Chinese Medicine. Ginsenosides are the major bioactive components. However, the shortage and high cost of some ginsenoside reference standards make it is difficult for quality control of P. ginseng. Objective: A method, single standard for determination of multicomponents (SSDMC), was developed for the simultaneous determination of nine ginsenosides in P. ginsen g (ginsenoside Rg1, Re, Rf, Rg2, Rb1, Rc, Rb2, Rb3, Rd). Materials and Methods: The analytes were separated on Inertsil ODS-3 C18 (250 mm × 4.6 mm, 5 μ m) with gradient elution of acetonitrile and water. The flow rate was 1 mL/min and detection wavelength was set at 203 nm. The feasibility and accuracy of SSDMC were checked by the external standard method, and various high-performance liquid chromatographic (HPLC) instruments and chromatographic conditions were investigated to verify its applicability. Using ginsenoside Rg1as the internal reference substance, the contents of other eight ginsenosides were calculated according to conversion factors (F) by HPLC. Results: The method was validated with linearity (r2 ≥ 0.9990), precision (relative standard deviation [RSD] ≤2.9%), accuracy (97.5%–100.8%, RSD ≤ 1.6%), repeatability, and stability. There was no significant difference between the SSDMC method and the external standard method. Conclusion: New SSDMC method could be considered as an ideal mean to analyze the components for which reference standards are not readily available. Abbreviations used: DRT: Different value of retention time; F: Conversion factor; HPLC: High-performance Liquid Chromatography; LOD: Limit of detection; LOQ: Limit of quantitation; PD: Percent difference; PPD: 20(S)-protopanaxadiol; PPT: 20(S)-protopanaxatriol; RSD: Relative standard deviation; SSDMC: Single Standard for Determination of Multicomponents; TCM: Traditional Chinese Medicine.
  2,945 173 3
Quality-by-Design: Multivariate model for multicomponent quantification in refining process of honey
Xiaoying Li, Zhisheng Wu, Xin Feng, Shanshan Liu, Xiaojie Yu, Qun Ma, Yanjiang Qiao
January-March 2017, 13(49):193-198
DOI:10.4103/0973-1296.196310  PMID:28216906
Objective: A method for rapid analysis of the refining process of honey was developed based on near-infrared (NIR) spectroscopy. Methods: Partial least square calibration models were built for the four components after the selection of the optimal spectral pretreatment method and latent factors. Results: The models covered the samples of different temperatures and time pointstherefore the models were robust and universal. Conclusions: These results highlighted that the NIR technology could extract the information of critical process and provide essential process knowledge of the honey refining process. Abbreviation used: NIR: Near-infrared; 5-HMF: 5-hydroxymethylfurfural; RMSEP: Root mean square error of prediction; R: correlation coefficients; PRESS: prediction residual error-sum squares; TCM: Traditional Chinese medicine; HPLC: High-performance liquid chromatography; HPLC-DAD: HPLC-diode array detector; PLS: Partial least square; MSC: multiplicative scatter correction; RMSECV: Root mean square error of cross validation; RPD: Residual predictive deviation; 1D: 1st order derivative; SG: Savitzky-Golay smooth; 2D: 2nd order derivative.
  2,969 109 1
Cucurbitane-type triterpenoids from the blood glucose-lowering extracts of Coccinia indica and Momordica balsamina fruits
Ujjwal Kaushik, Vidhu Aeri, R Mir Showkat, Mohammad Ali
January-March 2017, 13(49):115-121
DOI:10.4103/0973-1296.203991  PMID:28479735
Context: Few vegetables that are commonly consumed in India as part of diet have been claimed for their antidiabetic potential. Objective: The present study was aimed at evaluating preventive effects of cucurbit vegetables namely, Coccinia indica and Momordica balsamina belonging to family Cucurbitaceae in diabetic hyperglycemia. Materials and Methods: The fruits of M. balsamina and C. indica were extracted with chloroform and fractionated with hexane to prepare an extract rich in moderately polar components. These extracts were used for evaluating the effect of these cucurbits in nicotinamide/streptozotocin-induced type 2 diabetes. Streptozotocin–nicotinamide-induced diabetic animals were orally treated with chloroform extract of fruits (250 mg/kg BW) given daily for a week separately. Results: Both the extracts reduced fasting blood glucose significantly (P < 0.05 versus diabetic control) when estimated on seventh day of treatments. Pretreatment with fruit extracts for 7 days also blunted the OGTT (oral glucose tolerance test) curve. Results indicated that C. indica and M. balsamina fruits possess beneficial effects in diabetes by lowering elevated blood glucose level. Six cucurbitane-type triterpenoids were isolated from bioactive extracts of C. indica (1-3) and M. balsamina (4-6). The structures of these compounds were elucidated on the basis of spectroscopic data analysis.Conclusion: The study concludes that the inclusion of C. indica and M. balsamina fruits in food can be useful for newly diagnosed diabetic patients or highrisk group of population for glycemic control. Abbreviation used: C: indica (Coccinia indica), M: balsamina (Momordica balsamina), Kbr: Potassium bromide, FTIR: Fourier transform infrared spectroscopy, COSY: Corelated Spectroscopy, DEPT: Distortionless Enhancement by Polarization Transfer, DMSO: Dimethyl sulfoxide, TMS: tetramethylsilane, ESI-MS: Electrospray Ionization mass spectrometry, TLC: thin layer chromatography, STZ-NA: Streptozotocin-nicotinamide, CMC: carboxy methyl cellulose, BW: body weight, ANOVA: analysis of variance, AUC: area under curve
  2,872 203 1
Exploratory studies of (-)-Epicatechin, a bioactive compound of Phyllanthus niruri, on the antioxidant enzymes and oxidative stress markers in D-galactosamine-induced hepatitis in rats: A study with reference to clinical prospective
Bhasha Shanmugam, Kondeti Ramudu Shanmugam, Sahukari Ravi, Ganjikunta Venkata Subbaiah, Chilakala Ramakrishana, Korivi Mallikarjuna, Kesireddy Sathyavelu Reddy
January-March 2017, 13(49):56-62
DOI:10.4103/0973-1296.203973  PMID:28479727
Background: Hepatitis is a health problem affecting millions of people worldwide and it is the major risk factor for liver cirrhosis. In India, many plants are used to treat hepatitis. But little is known about the effects of (-)-epicatechin a bioactive compound of Phyllanthus niruri (PN) in hepatitis rats. Objective: The present study was designed to explore the antioxidant property of (-)-epicatechin isolated from PN in D-Galactosamine (D-GalN) induced hepatitis rats. Materials and Methods: The rats are divided into five groups as per the experimental design. (-)-Epicatchin pretreatment was given to the hepatitis rats for 21 days and biochemical analysis was carried out. The hepatic antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR), glutathione S-transferase (GST), reduced glutathione (GSH), and malondialdehyde (MDA) and serum markers aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), albumin, and bilirubin are estimated. Results: All the antioxidant enzymes activities and albumin levels are depleted in hepatitic rats. Whereas GST, ALP, AST, ALT activities and MDA, and bilirubin levels are elevated in hepatitis rats, (-)-epicatechin pretreatment increased all the antioxidant enzymes and decreased the GST, ALP, AST, ALT, and MDA levels in hepatitis rats. However, histopatholoigic studies also proves that (-)-epicatechin pretreatment decreased the tissue damage in hepatitis condition. This is the first report on the antioxidant enzymes and hepatoprotective effect of (-)-epicatechin in hepatitis rats. Conclusion: From this study, we conclude that (-)-epicatechin treatment decreased the oxidative damage in hepatitis rats. Abbreviations used: Phyllanthus niruri (PN), D-Galactosamine (D-GalN),superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione reductase (GR), glutathione s transferase (GST), reduced glutathione (GSH) and malondialdehyde (MDA) aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), World Health Organisation (WHO), Indian Institute of Science (IISc), Nicotinamide adenine dinucleotide phosphate (NADPH), thiobarbituric acid reactive substances (TBARS).
  2,824 225 14
Ginger treatment ameliorates alcohol-induced myocardial damage by suppression of hyperlipidemia and cardiac biomarkers in rats
Ganjikunta Venkata Subbaiah, Korivi Mallikarjuna, Bhasha Shanmugam, Sahukari Ravi, Patan Usnan Taj, Kesireddy Sathyavelu Reddy
January-March 2017, 13(49):69-75
DOI:10.4103/0973-1296.203891  PMID:28479729
Background: Alcohol-induced hyperlipidemia is positively correlated with cardiovascular diseases. Several herbal extracts have been reported to protect the cardiac injury and suppress the hyperlipidemia. However, the effect of ginger extracts on alcohol-induced hyperlipidemia and associated myocardial damage remains unclear. Objective: This study investigated the cardio-protective properties of ginger ethanolic extract (Gt) against alcohol-induced myocardial damage, and further distinguished the association between hyperlipidemia and occurrence of myocardial damage in rats. Materials and Methods: Twenty four Wistar male albino rats (250 ± 20 g) were divided into four groups including, Normal control (NC) (0.9% NaCl), Ginger treated (Gt) (200 mg/Kg b.w.), Alcohol treated (At) (20% of 6g/kg b.w. alcohol), and Alcohol along with Ginger treatment (At+Gt). In this study, lipid profiles such as fatty acids, triglycerides, total cholesterol, phospholipids, low density lipoprotein and high density lipoproteins, and cardiac biomarkers, including LDH, AST, CK-MB, cTn-T and cTn-I were examined in rats. Furthermore, histopathological studies were also conducted. Results: We found that alcohol-induced myocardial damage was associated with increased lipid profile except high density lipoprotein in alcohol treated (20%, 6g/kg b.w.) rats compared with control. Ginger treatment significantly reduced the alcohol-induced lipid profiles except high density lipoproteins. Furthermore, elevated cardiac biomarkers activity with alcohol intoxication was substantially suppressed by ginger treatment. In addition, ginger treatment for 7-weeks significantly minimized the alcohol-induced myocardial damage. Conclusion: Our results concluded that ginger could protect alcohol-induced myocardial damage by suppression of hyperlipidemia and cardiac biomarkers. Abbreviation Used:Gt: Ginger Ethanolic Extract; NC: Normal Control; At: Alcohol treated; MI: Myocardial Infarction
  2,852 179 7
LETTER TO THE EDITOR
Nutraceutical value of pure curcumin
Biji T Kurien, Hiroyuki Matsumoto, R Hal Scofield
January-March 2017, 13(49):161-163
DOI:10.4103/0973-1296.203988  PMID:28479742
  2,809 139 4
ORIGINAL ARTICLES
Wheatgrass extract ameliorates hypoxia-induced mucin gene expression in A549 cells
Ju hwan Sim, Moon-Hee Choi, Hyun-Jae Shin, Ji-Eun Lee
January-March 2017, 13(49):7-12
DOI:10.4103/0973-1296.197660  PMID:28216876
Background: Wheatgrass is known to have antioxidant, antiaging, and anti-inflammatory effect. However, its protective effect against hypoxia is not yet evaluated. Objective: In this study, we evaluated the protective and anti-inflammatory effect of wheatgrass against the hypoxia in airway epithelial cells. Materials and Methods: A549 human lung adenocarcinoma cells were incubated in a hypoxic condition (CO2 5%/O2 1%) for 24 hr in the presence of different concentration of wheatgrass 50, 75, 100, and 150 μg/mL, and the magnitude of each immunologic response produced by the A549 cells was compared. The mRNA expression level of mucin gene (MUC), 5A, 5B, 8, GM-CSF, TNF-α, and VEGF were evaluated by using real-time polymerase chain reaction. The MUC proteins level before and after knocking out the hypoxia-inducible factor (hif)-1α via short interfering (si) RNA transfection were assessed by immunoblot analysis. Accordingly, the involved cell signaling pathway was evaluated by immunoblot analysis. Results: The inflammatory cytokines (GM-CSF, TNF- α) and the expressions of MUC 5A, 5B, and 8 were augmented by hypoxia. The augmented MUC expression was decreased by the wheatgrass extract administration. Hif-1α gene expression after hypoxia exposure was decreased by wheatgrass. Knockdown of hif-1α by siRNA reduced the mucin gene expression and which was more enhanced by wheatgrass extract. Conclusion: Theses results suggest that wheatgrass may be useful in the treatment of sinonasal disease by inhibiting mucus hypersecretion in airway epithelium. Abbreviations used: A549: human lung adenocarcinoma cells, GM-CSF: granulocyte-macrophage colony stimulating factor, HIF: hypoxia inducible factor, IL: interleukin, MUC: mucin, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, TNF: tumor necrosis factors, VEGF: vascular endothelial growth factor, si RNA: short interfering RNA
  2,833 106 -
Anti-hepatocarcinogenic and anti-oxidant effects of mangrove plant Scyphiphora hydrophyllacea
Sameera R Samarakoon, Chanthirika Shanmuganathan, Meran K Ediriweera, Poorna Piyathilaka, Kamani H Tennekoon, Ira Thabrew, Prasanna Galhena, E Dilip De Silva
January-March 2017, 13(49):76-83
DOI:10.4103/0973-1296.203989  PMID:28479730
Context: Scyphiphora hydrophyllacea is a shrub mangrove plant of the family Rubiaceae and not yet been studied for anti-hepatocarcinogenic effects. Objectives: We investigated possible in vitro anti-hepatocarcinogenic and antioxidant properties of S. hydrophyllacea. Materials and Methods: Dried leaves of S. hydrophyllacea were sequentially extracted into hexane, chloroform, ethyl acetate, and methanol and tested for cytotoxicity on HepG2 cells by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and sulforhodamine B assays, and for antioxidant activities by the free radical 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) assays. Total phenolic and flavonoid contents were estimated in all four extracts. The hexane and chloroform extracts were tested for pro-apoptotic properties in HepG2 cells, and bioactive components were identified by gas chromatography–mass spectrometry (GC-MS) analysis. Results: The hexane and chloroform extracts showed dose-dependent and time-dependent cytotoxic effects. Morphological changes observed under fluorescence microscope related to apoptosis, and significant (P < 0.001) increases in caspase 3 and 9 levels were observed in hexane and chloroform extract-treated cells. Slight DNA fragmentation was observed only in response to the chloroform extract. mRNA expressions of p53 and Bax were significantly upregulated by low doses of hexane and chloroform extracts. Highest antioxidant activity was observed in the methanol extract. GC-MS profiles identified 24 and four major compounds in the hexane and chloroform extracts, respectively. These included some known anticancer compounds such as lupeol. Conclusion: Cytotoxicity, antioxidant effects, and apoptosis-related changes exerted by hexane and chloroform extracts of S. hydrophyllacea concluded that these two extracts are good source for isolation of possible anticarcinogenic compounds. Abbreviation used: DPPH: 1,1-diphenyl-2-picryl-hydrazyl, ABTS: 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid, GC-MS: gas chromatography–mass spectrometry, DNA: deoxyribonucleic acid, HCC: Hepatocellular carcinoma, GAE: gallic acid equivalents, SRB: sulforhodamine B, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, AO/EB: acridine orange/ethidium bromide, GAPDH: Glyceraldehyde 3-phosphate dehydrogenase, IC50: half maximal inhibitory concentration; QE: quercetin equivalents, HE: hexane extract, CE: chloroform extract, EAE: ethyl acetate extract, ME: methanolic extract, TPC: total polyphenol content, TFC: total flavonoid content, ANOVA: Analysis of variance
  2,762 175 3
Antioxidant and nephroprotective activities of the extract and fractions of Homonoia riparia Lour
Seena kanniparambil Xavier, Shoja Muhammed Haneefa, Devkar Raviraj Anand, Picheswara Rao polo, Rajalekshmi Maheshwari, Chandrashekara Shastry Shreedhara, Manganahalli Manjunath Setty
January-March 2017, 13(49):25-30
DOI:10.4103/0973-1296.197647  PMID:28216879
Background: Homonoia riparia is a plant, which is widely used in the indigenous system of medicine for the treatment of urolithiasis, renal disorders and inflammatory conditions. This is the first report on the antioxidant and nephroprotective activities of whole plant of H. riparia. Objective: The present study aims at investigating the in vitro antioxidant and nephroprotective activity of the methanol extract and its different fractions of H. riparia. Methods: Petroleum ether (HRPE), Ethyl acetate (HREA), Butanol (HRBU), aqueous fractions (HRAQ) were prepared from the crude methanol extract of H. riparia (HRM) using liquid partitioning. Total phenolic content, flavonoid content and antioxidant activity assay were performed according to suitable methods. Nephroprotective activities were evaluated by MTT assay using Human Embryonic Kidney cells against cisplatin induced toxicity. Quantification of gallic acid was performed using validated HPTLC method. Results: The studies showed that extract and fractions possess significant nephroprotective activity against cisplatin induced renal toxicity. All the extracts/fractions of whole plant of Homonoia riparia was found to be significantly reducing cisplatin induced toxicity (< 0.05). The highest activity was observed with HRBU and HRAQ with a percentage viability of 293.09 ± 4.3 and 345.07 ± 3.2 at a concentration of 200 µg/ml. Gallic acid was detected in the HRM/fractions using HPTLC. Abbreviations used: HPTLC: High Performance Thin Layer Chromatography, DPPH: 1,1-diphenyl-2-picrylhydrazyl, ABTS: 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, MTT: 3-(4,5-dimethylthiazolyl-2-yl)-2,5- diphenyl tetrazolium bromide, GAE: Gallic acid equivalents, QE: Quercetin equivalents, HEK: Human Embryonic Kidney, HRM: Methanol extract of H. riparia, HRPE: Petroleum ether fraction of H. riparia, HREA: ethyl acetate fraction of H. riparia, HRBU: Butanol fraction of H.riparia, HRAQ: Aqueous fraction of H. riparia, DMEM: Dulbecco's minimum essential medium, FBS: Foetal bovine serum, DMSO: Dimethyl sulfoxide, ANOVA: One way analysis of variance.
  2,755 110 -
In vitro Toxicity and genotoxic activity of aqueous leaf extracts from four varieties of Olea europea (L)
Luc Verschaeve, Hayet Edziri, Roel Anthonissen, Dalenda Boujnah, Fethia Skhiri, Hechmi Chehab, Mahjoub Aouni, Maha Mastouri
January-March 2017, 13(49):63-68
DOI:10.4103/0973-1296.203980  PMID:28479728
Aim: Despite its therapeutic value almost nothing is known about potential adverse health effects of Olea europea L. We therefore investigated the in vitro toxicity and genotoxicity of leaf extracts of this plant. Material and Methods: Extracts from olive tree leaves were obtained from four different regions in Tunisia. We investigated the in vitro toxicity, genotoxicity and antigenotoxicity of their aqueous extracts using the neutral red (NR) uptake, Vitotox and alkaline comet assays. Results: None of the extracts were found to be toxic and none of them were genotoxic, although some doubt exists for the extract obtained at Meski (North of Tunisia). On the basis of the Vitotox test only, none of the extracts appeared to have antigenotoxic (or cogenotoxic) properties. Discussion: The negative genotoxicity underline the safe use of the leaves, for example, as hypoglycemic and antidiabetic preparations. Lack of antigenotoxicity may indicate that the previously reported anticancer effects do not result from protection against genotoxicity. Abbreviation list: BaP : benzo(α)pyrene, EMS: ethyl methane sulfonate, LMP: low melting point, NI50: 50% inhibition of NRU, NR: neutral red, NRU: neutral red uptake, OD: optical density, PBS: phosphate buffer saline, SDS: sodium dodecyl sulphate, S/N: signal to noise ratio, 4NQO : 4-nitroquinoline oxide
  2,688 132 5
Antiproliferative and proapoptotic activities of marine sponge Hyrtios erectus extract on breast carcinoma cell line (MCF-7)
Ramachandran Muthiyan, Balwin Nambikkairaj, Nilkamal Mahanta, Titus Immanuel, Rahul Shubhra Mandal, Kubendiran Kumaran, Arun Kumar De
January-March 2017, 13(49):41-47
DOI:10.4103/0973-1296.203983  PMID:28479725
Background: Marine sponge is a rich natural resource of many pharmacologically important compounds. Objective: Marine sponge Hyrtios erectus, collected from North Bay, South Andaman Sea, India, was screened for potential antiproliferative and proapoptotic properties on a breast adenocarcinoma cell line (MCF-7). Materials and Methods: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to test the antiproliferative and cytotoxicity effects of the sponge extract. Analysis of apoptosis and cell cycle stages were done by flow cytometry. The expression of several apoptotic-related proteins in MCF-7 cells treated by the extract was evaluated by Western blot analysis. Various analytical techniques including Fourier transform infrared spectroscopy, gas chromatography-mass spectrometry, and nuclear magnetic resonance were employed to determine the identity of the active compounds in the sponge extract. Results: N-Hexane extract of the sponge inhibited proliferation of the MCF-7 cell line in a dose- and time-dependent manner. Exposure of the sponge extract triggered apoptosis of the MCF-7 cells, induced DNA fragmentation, and arrested the cells in G2/M phase. Treatment of the sponge extract induced downregulation of antiapoptotic Bcl-2 protein and upregulation of Bax, caspase-3, caspase-9, and fragmented poly(ADP ribose)polymerase proteins in MCF-7 cells. Five bioactive compounds have been identified in the extract. Conclusion: The antiproliferative and proapoptotic activities of the tested extract suggested the pharmacologic potential of the identified compounds. Further characterization of the identified compounds are in progress. Abbreviations used: GC-MS: Gas chromatography-mass spectrometry; FT-IR: Fourier transform infrared spectroscopy; NMR: Nuclear magnetic resonance; MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.
  2,635 166 2
Technological evaluation of emulsions containing the volatile oil from leaves of Plectranthus Amboinicus Lour
Pablo Queiroz Lopes, Fabíola Bernardo Carneiro, Ana Braz Letícia de Sousa, Sócrates Golziodo Santos, Elquio Eleamen Oliveira, Luiz Alberto Lira Soares
January-March 2017, 13(49):159-167
DOI:10.4103/0973-1296.197646  PMID:28216901
Background: Plectranthus amboinicus Lour is a species which is widespread throughout tropical countries where it is widely used against respiratory tract disorders such as bronchodilator, antitussive, and expectorant conditions. Objective: This study aims to characterize the essential oil of P. amboinicus (PaEO) and produce and evaluate emulsions containing PaEO. Materials and Methods: The essential oil was characterized by physical-chemical analyses for density, refractive index, 90% ethanol solubility, color, appearance, and identification by gas chromatography coupled to mass spectrometry detection. The emulsions were prepared following a hydrophile-lipophile balance [HLB] spreadsheet design from two nonionic surfactants (Span 80® and Tween 20®) producing HLB values ranging from 4.3 to 16.7. The products were stored at room temperature at 5°C. The emulsion stabilities were tested both in the long and short-term.Results: The PaEO was obtained by steam distillation and the total extraction was reached after 3 hours yielding of 0.2% (w/w). This essential oil was characterized by physicochemical analyses for density [1.5 g.ml-1], refraction index [0.9167], ethanol 90% solubility [1:2], color, and appearance (yellow/clear). Nineteen components were identified in the oil, among them the sesquiterpenes: carvacrol [33.50%], p-cymene [28.20%] and γ-terpinene [14.77%]. The emulsions obtained successfully showed, for the first time, HLB values for essential oils from Plectranthus amboinicus [15.7].Conclusion: The experimental data shows a relationship between HLB values of the surfactant mixtures contributing to the emulsified systems production containing phytopharmaceuticals. Such an approach is of great importance to the development of lipid carriers for therapeutic drugs. Abbreviations used: PaEO: essential oil of P. amboinicus, HLB: hydrophilic-lipophilic balance, CI: Creaming Index, MET: micro-emultocrit technique
  2,635 106 -
Green biosynthesis, characterization, in vitro antidiabetic activity, and investigational acute toxicity studies of some herbal-mediated silver nanoparticles on animal models
Kalakotla Shanker, Gottumukkala Krishna Mohan, Md Ashwaq Hussain, Naradala Jayarambabu, Poka Lakshmi Pravallika
January-March 2017, 13(49):188-192
DOI:10.4103/0973-1296.197642  PMID:28216905
Diabetes is a metabolic disorder characterized by hyperglycemia, altered carbohydrate, lipid and protein metabolism. In recent studies, Nanoscience and nanotechnology are blazing fields for researchers; for researchers; of late there has been a prodigious excitement in the field of nanopharmacology to study silver nanoparticle (SNP) synthesis using natural products. Biological methods have been used to synthesize SNPs using medicinally active plants having an antidiabetic role, and this made us to assess the biologically synthesized SNPs from the seed extract of Psoralea corylifolia using 1 mM silver nitrate solution. The synthesized herbal&-mediated SNPs (HMSNPs) were subjected to various characterization techniques such as X&-ray diffraction analysis (XRD), energy dispersive X&-ray (EDX) analysis, transmission electron microscope (TEM), and differential light scattering (DLS), respectively. In the current study the HMSNPs were tested to observe the in vitro antidiabetic activity and possible toxic effects in healthy female albino mice by following OECD guidelines&-425. Huge data from biochemical, cellular, mouse, and chemical inhibitor studies have recognized protein tyrosine phosphatase 1B (PTP1B) as a major negative regulator of insulin signaling. In addition, corroboration suggests that insulin action can be enhanced by the inhibition of PTP1B. Keeping in view of the above fact, the PTP1B assay was done to determine the PTP1 B inhibitory effect of HMSNPs. It can be concluded that medicinal plants can be a good source for the synthe sis of HMSNPs. This study can be used for the development of valuable nanomedicines to treat various ailments, and it also highlights the safety and biocompatibility of SNPs within a biological cell; in vivo parameters need to be considered for further discoveries. Abbreviations used: HMSNPs: Herbal mediated silver nanoparticles, XRD: X&-ray diffraction, EDX: Energy dispersive X&-ray analysis, TEM: Transmission electron microscope, PTP1B: Protein tyrosine phosphotase 1B, OECD: Organization for economic cooperation and development
  2,541 158 -
Anti-nociceptive effect of 7-methoxy coumarin from Eupatorium Triplinerve vahl (Asteraceae)
Binoy Varghese Cheriyan, Parimala Kadhirvelu, Jagan Nadipelly, Jaikumar Shanmugasundaram, Vijaykumar Sayeli, Viswanathan Subramanian
January-March 2017, 13(49):81-84
DOI:10.4103/0973-1296.197650  PMID:28216887
Aim: To evaluate the anti-nociceptive activity of 7-methoxy coumarin isolated from ethyl acetate fraction of the alcoholic extract of Eupatorium triplinerve Vahl. Materials and Methods: The shade dried leaves of E. triplinerve were extracted with ethyl alcohol and the extract was condensed. This extract was fractionated with n-hexane, ethyl acetate, and n-butanol. The ethyl acetate fraction was subjected to column chromatography which yielded a crystalline compound-A, which was investigated for spectral characteristics. Pharmacological studies: The isolated compound-A was subjected to behavioral studies and anti-nociceptive evaluation in mice by acetic acid induced writhing and formalin induced nociception. Results: The spectral studies indicated that the structure of compound-A complies with 7- methoxy coumarin. Pre-treatment with 7-methoxy coumarin reduced the number of abdominal constrictions in mice and decreased the time spent in paw licking and biting response in formalin assay. There were no significant behavioral changes. Conclusion: A dose dependent anti-nociceptive action of 7- methoxy coumarin was revealed by the present experiments which support the traditional use of E. triplinerve in pain and inflammatory disorders. Abbreviation used: TLC-Thin layer chromatography, Kg-kilogram, g-gram, TXB2-Thromboxane B2, UV-Ultraviolet, IgE-Immunoglobulin E, s.c-subcutaneous, p.o-oral route
  2,532 111 -
The inhibition of hepatic and renal glucuronidation of p-Nitrophenol and 4-Methylumbelliferone by oil palm empty fruit bunch lignin and its main oxidation compounds
Norliyana Mohamad Salleh, Sabariah Ismail, Mohamad Nasir Mohamad Ibrahim
January-March 2017, 13(49):102-114
DOI:10.4103/0973-1296.203990  PMID:28479734
Background: In order to develop oil palm empty fruit bunch (EFB) lignin as a nutraceutical and health supplement, the investigation of its potential in interacting with other drugs via inhibition of drug-metabolizing enzymes (DMEs) would ensure product safety. Objective: The study was aimed to investigate the in vitro effect of oil palm EFB lignin and its main oxidation compounds on phase II DME UDP-glucuronosyltransferases (UGTs) in rat liver and kidney microsomes. Materials and Methods: The p-nitrophenol (p-NP) and 4-methylumbelliferone (4-MU) were employed as probe substrates in glucuronidation assays. The effect of soda oil palm EFB lignin on Vmax, Km, CLint, Ki, and mode of inhibition of 4-MU glucuronidation in RLM was also determined. Results: The inhibitory potency of oil palm EFB lignin for both p-NP and 4-MU glucuronidation in rat liver microsome (RLM) and rat kidneys microsomes (RKM) was found to be in the rank order of soda > kraft > organosolv. However, the inhibitory potency of its main oxidation compounds were in the rank order of vanillin > syringaldehyde > p-hydroxybenzaldehyde. Soda oil palm EFB lignin exhibited mixed-type inhibition against 4-MU glucuronidation in RLM, showing the change in apparent Vmax and with only a minor effect on Km compared with control. Conclusions: The findings showed that effect of oil palm EFB lignin on both p-NP and 4-MU glucuronidation in RLM and RKM was enhanced by the presence of vanillin as well as flavonoids. Kinetic study showed that soda oil palm EFB lignin exhibited strong inhibition on UGT activity in RLM with mixed-type inhibition mode. Abbreviations used: p-NP: p-Nitrophenol, 4-MU: 4-Methylumbelliferone, EFB: Empty fruit bunch, DME: Drug-metabolizing enzymes, UGT: UDP-glucuronosyltransferase, Vmax: Maximal reaction velocity, Km: Michaelis-Menten constant, CLint: Intrinsic clearance, Ki: Dissociation constant of an inhibitor enzyme complex, 4-MUG: 4-Methylumbelliferone glucuronide, DMSO: Dimethyl sulfoxide, IC50: Half maximal inhibitory concentration, p-NPG: p-Nitrophenol glucuronide, RKM: Rat kidneys microsomes, RLM: Rat liver microsome, UDPGA: UDP-glucuronic acid, TCA: trichloroacetic acid, MPA: mycophenolic acid
  2,479 157 1
Antiprotozoal constituents from Annona cherimola Miller, a plant used in mexican traditional medicine for the treatment of diarrhea and dysentery
Fernando Calzada, Jose Correa-Basurto, Elizabeth Barbosa, David Mendez-Luna, Lilian Yepez-Mulia
January-March 2017, 13(49):148-152
DOI:10.4103/0973-1296.197636  PMID:28216899
Background:Annona cherimola Miller (Annonaceae) is a medicinal plant frequently recommended in Mexican traditional medicine for the treatment of gastrointestinal disorders such as diarrhea and dysentery.Objective: This work was undertaken to obtain information that support the traditional use of A. cherimola, on pharmacological basis using in vitro and computational experiments.Material and Methods: Bioassay-guided fractionation of the ethanol extract of the leaves of A. cherimola afforded five phenolic compounds:caffeic acid, quercetin, kaempferol, nicotinflorin, and rutin.Results: The in vitro antiprotozoal assay showed that kaempferol was the most potent antiamoebic and antigiardial compound with IC50 values of 7.9 μg/mL for Entamoeba histolytica and 8.7 μg/mL for Giardia lamblia. Computational molecular docking study showed that kaempferol interacted in a region different than metronidazole in the enzyme pyruvate:ferredoxin oxidoreductase (PFOR).Conclusion: Considering that PFOR is a target of metronidazole; kaempferol may be a lead compound for the development of novel antiprotozoal agent. Also, these findings give support to the use of A. cherimola in the traditional medicine from México for the treatment of diarrhea and dysentery. Abbreviations used: PFOR:Pyruvate:ferredoxin oxidoreductase, G:lamblia:Giardia lamblia, E:histolytica:Entamoeba histolytica
  2,503 131 -
Anticancer potential of nutraceutical formulations in MNU-induced mammary cancer in Sprague Dawley rats
Gummalla Pitchaiah, Annapurna Akula, Vishala Chandi
January-March 2017, 13(49):46-50
DOI:10.4103/0973-1296.197652  PMID:28216882
Background: Nutraceuticals help in combating some of the major health problems of the century including cancer, and 'nutraceutical formulations' have led to the new era of medicine and health.Objective: To develop different nutraceutical formulations and to assess the anticancer potential of nutraceutical formulations in N-methyl-N-nitrosourea (MNU)-induced mammary cancer in Sprague Dawley rats. Materials and Methods: Different nutraceutical formulations were prepared using fine powders of amla, apple, garlic, onion, papaya, turmeric, and wheat grass with and without cow urine distillate. Total phenolic content, acute oral toxicity, and microbial load of nutraceutical formulations were assessed. The anticancer potential of nutraceutical formulations was evaluated against MNU-induced mammary cancer in female Sprague Dawley rats. Results: Improvement in total phenolic content was significant (P < 0.001) after self-fortification process. Toxicity studies showed that the nutraceutical formulations were safe to use in animals. Microbial load was within the limits. Significant longer tumor-free days (P < 0.01), lower tumor incidence (P < 0.01), lower tumor multiplicity (P < 0.05) and tumor burden (P < 0.01) were observed for nutraceutical formulation-treated groups. Conclusion: Combination of whole food-based nutraceuticals acted synergistically in the prevention of mammary cancer. Further, the process of fortification is novel and enhanced the anticancer potential of nutraceutical formulations. Abbreviations used: HMNU: N-methyl-N-nitrosourea, CAM: Complementary and Alternative Medicine, NF: Nutraceutical Formulation, SFNF: Self-Fortitfied Nutraceutical Formulation, NFCUD: Nutraceutical Formulation fortified with Cow Urine Disstillate, SFNFCUD: Self-Fortified Nutraceutical Formulation fortified with Cow Urine Disstillate, CPCSEA: Committee for the Purpose of Control and Supervision of Experiments on Animals, OECD: Organisation for Economic Co-operation and Development, TPC: Total Phenolic Content, ANOVA: Analysis of Variance, GAE: Gallic Acid Equivalent, cfu/g: Colony forming unit per g
  2,436 118 -
Nardostachys jatamansi root extract modulates the growth of IMR-32 and SK-N-MC neuroblastoma cell lines through MYCN mediated regulation of MDM2 and p53
Snehal Suryavanshi, Prerna Raina, Rashmi Deshpande, Ruchika Kaul-Ghanekar
January-March 2017, 13(49):21-24
DOI:10.4103/0973-1296.197645  PMID:28216878
Aim: The present study evaluated the effect of ethanolic extract of Nardostachys jatamansi roots (NJet) on MYCN mediated regulation of expression of MDM2 and p53 proteins in neuroblastoma cell lines, IMR-32 and SK-N-MC. Materials and Methods: The effect of NJet on cell viability was determined by MTT; and on growth kinetics was evaluated by trypan blue dye exclusion method and soft agar assay. The expression of p53, MDM2 and MYCN proteins in response to NJet treatment was evaluated by immunoblotting. Results: NJet decreased the viability of neuroblastoma cells without affecting the viability of non-cancerous, HEK-293 cells. It altered the growth kinetics of the cancer cells in a dose-dependent manner. NJet down regulated the expression of MYCN and MDM2 proteins with a simultaneous increase in the expression of tumor suppressor protein p53. Conclusions: The present data demonstrated that NJet regulated the growth of IMR-32 and SK-N-MC through reduction in MYCN expression that lead to down regulation of MDM2 protein and increase in p53 expression. These preliminary results warrant further in depth studies to explore the therapeutic potential of Nardostachys jatamansi in the management of neuroblastoma. Abbreviations used: NJet: N. jatamansie thanolic extract; MTT: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenylthiazolium bromide; FBS: Fetal bovine serum; FITC: Fluorescein isothiocyanate
  2,417 129 -
Cytotoxic metabolites from Callyspongia siphonella display antiproliferative activity by inducing apoptosis in HCT-116 cells
Tariq R.A. Sobahi, Seif-Eldin N Ayyad, Ahmed Abdel-Lateff, Mardi M Algandaby, Hajer S Alorfi, Ashraf B Abdel-Naim
January-March 2017, 13(49):37-40
DOI:10.4103/0973-1296.203970  PMID:28479724
Objectives: To evaluate the antiproliferative effect of the isolated metabolites from Callyspongia siphonella. Methods: Different chromatographic methods have been done on the organic extract of the marine sponge aiming at isolating the bioactive metabolites. The cytotoxicity of the isolated compounds has been evaluated against the human colorectal cancer cell line; HCT-116, employing SRB assay. The flow cytometry assay was applied to measure the cell cycle analysis. Results: Six metabolites (1–6) were obtained. The compounds 4–6 exhibited IC50 values (µM ± SD) of 95.80± 1.34, 14.8 ± 2.33, and 19.8 ± 3.78, respectively. Cell cycle distribution analysis revealed that sipholenol A (5) and sipholenol L (6) induced G2/M and S phase arrest with concomitant increase in the pre-G cell population. Furthermore, 5 and 6 increased the nuclear expression of the pro-apoptotic protein-cleaved caspase-3 that effectively drives cellular apoptosis via caspase-3-dependent pathway. Conclusions: The antiproliferative activity of 5 and 6 can be recognized, at least partly, due to their ability to induce cellular apoptosis. Abbreviations used: A549 (human lung carcinoma), Caco-2 (Human ColonCarcinoma), CHCl3 (Chloroform), HCT 116 (Human Colon Carcinoma), HepG2 (Liver Hepatocellular Carcinoma), HT-29 (Human Colorectal Adenocarcinoma), MCF-7 (Michigan Cancer Foundation-7; Human Breast Adenocarcinoma), MeOH (Methanol), NMR Nuclear Magnetic Resonance), PBS (Phosphate Buffered Saline), PC-3 (Human Prostate Cancer), PTLC (Preparative Thin Layer Chromatography), RPMI-1640 (Roswell Park Memorial Institute medium), TLC (ThinLayer Chromatography).
  2,383 142 5
Effect of absolute from Hibiscus syriacus L. flower on wound healing in keratinocytes
Seok Won Yoon, Kang Pa Lee, Do-Yoon Kim, Dae Il Hwang, Kyung-Jong Won, Dae Won Lee, Hwan Myung Lee
January-March 2017, 13(49):85-89
DOI:10.4103/0973-1296.197644  PMID:28216888
Background: Proliferation and migration of keratinocytes are essential for the repair of cutaneous wounds. Hibiscus syriacus L. has been used in Asian medicine; however, research on keratinocytes is inadequate. Objective: To establish the dermatological properties of absolute from Hibiscus syriacus L. flower (HSF) and to provide fundamental research for alternative medicine. Materials and Methods: We identified the composition of HSF absolute using gas chromatography-mass spectrometry analysis. We also examined the effect of HSF absolute in HaCaT cells using the XTT assay, Boyden chamber assay, sprout-out growth assay, and western blotting. We conducted an in-vivo wound healing assay in rat tail-skin. Results: Ten major active compounds were identified from HSF absolute. As determined by the XTT assay, Boyden chamber assay, and sprout-out growth assay results, HSF absolute exhibited similar effects as that of epidermal growth factor on the proliferation and migration patterns of keratinocytes (HaCaT cells), which were significantly increased after HSF absolute treatment. The expression levels of the phosphorylated signaling proteins relevant to proliferation, including extracellular signal-regulated kinase 1/2 (Erk 1/2) and Akt, were also determined by western blot analysis. Conclusion: These results of our in-vitro and ex-vivo studies indicate that HSF absolute induced cell growth and migration of HaCaT cells by phosphorylating both Erk 1/2 and Akt. Moreover, we confirmed the wound-healing effect of HSF on injury of the rat tail-skin. Therefore, our results suggest that HSF absolute is promising for use in cosmetics and alternative medicine. Abbreviations used: HSF: Hibiscus syriacus L. flower, Erk 1/2: extracellular signal-regulated kinase 1/2, EGF: epidermal growth factor, GC/MS: gas chromatography-mass spectrometry, DMEM: dulbecco's modified eagle medium, FBS: fetal bovine serum, BSA: bovine serum albumin, p-Akt: phosphorylation of Akt, p-Erk 1/2: phosphorylation of Erk 1/2
  2,372 122 -
Fatty acid profiling and in vitro antihyperglycemic effect of Leucas cephalotes (Roth) spreng via carbohydrate hydrolyzing enzyme inhibition
Anjali Verma, Anil Kumar, Dalip Kumar Upreti, Veena Pande, Mahesh Pal
January-March 2017, 13(49):22-25
DOI:10.4103/0973-1296.203993  PMID:28479721
Background: Leucas cephalotes has been used by many tribes to treat variety of diseases and known to have many essential secondary metabolites. To the best of our knowledge, it is the first comparative analysis of total fatty acid (FA) composition and α-amylase inhibition activity of L. cephalotes. Objective: The present study is carried out to explore the antihyperglycemic activity and FA contents of all parts of L. cephalotes. Material and Method: Fruits, leaves, stems, and roots part of L. cephalotes have been extracted in ethanol. Simultaneously, all plant parts have been extracted in hexane with Soxhlet extraction. Ethanolic extracts have been evaluated for antihyperglycemic activity and hexane extract have been analyzed for FA identification. Result: The present study indicated that ethanolic extract of fruit and leaves have shown significant α-amylase inhibitory activity with IC50 value of 92.86 ± 0.89 and 98.09 ± 0.69 μg/mL, respectively. FA composition of all the parts of L. cephalotes was analyzed by GC/MS. Nineteen FAs have been identified in all parts of L. cephalotes in which palmitic acid, oleic acid, linolenic acid, and linoleic acid were major FAs. Conclusion: The study indicates that L. cephalotes has significant potential to inhibit α-amylase enzyme and it is a rich source of essential FAs. Abbreviations used: DM: Diabetes Mellitus, FA: Fatty Acid, FFAs: Free Fatty Acids, FAME: Fatty Acid Methyl Ester, IC50: Inhibitor Concentration, GC-MS: Gas ChromatographyMass Spectrophotometer
  2,366 109 1
Isolation and characterisation of a proanthocyanidin with antioxidative, antibacterial and anti-cancer properties from fern Blechnum orientale
How-Yee Lai, Yau-Yan Lim, Kah-Hwi Kim
January-March 2017, 13(49):31-37
DOI:10.4103/0973-1296.197659  PMID:28216880
Background:Blechnum orientale Linn. (Blechnaceae), a fern, is traditionally used in the treatment of various ailments, such as skin diseases, stomach pain, urinary bladder complaints, and also as a female contraceptive. Previously, we reported a strong radical scavenging activity, antibacterial activity and cytotoxicity against HT29 colon cancer cells by aqueous extract of B. orientale. Objective: In this study, we attempted to isolate and identify the active compound from the aqueous extract of B. orientale. Materials and Methods: Aqueous extract of B. orientale was subjected to repeated MCI gel chromatography, Sephadex-LH-20, Chromatorex C18 and semi-preparative high performance liquid chromatography and was characterized using nuclear magnetic resonance and electrospray ionization mass-spectrometry spectroscopic methods. Antioxidant activity was determined using 2, 2-diphenyl-1-picrylhydrazyl radical scavenging assay. Antibacterial assays were conducted using disc diffusion whereas the minimum inhibitory concentration (MIC) and minimum bactericidal concentration were determined using the broth microdilution assay. Cytotoxicity was assessed using thiazolylblue tetrazoliumbromide. Results: A polymeric proanthocyanidin consisting of 2-12 epicatechin extension units and epigallocathecin terminal units linked at C4-C8 was elucidated. Bioactivity studies showed strong radical scavenging activity (IC50 = 5.6 ± 0.1 µg/mL), antibacterial activity (MIC = 31.3-62.5 µg/mL) against five gram-positive bacteria and selective cytotoxicity against HT29 colon cancer cells (IC50 = 7.0 ± 0.3 µg/mL). Conclusion: According to our results, the proanthocyanidin of B. orientale demonstrated its potential as a natural source of antioxidant with antibacterial and anti-cancer properties. Abbreviation used: CC: Column chromatography, DP: degree of polymerization, DPPH: 2,2-diphenyl-1-picrylhydrazyl, ESI-MS: electronsprayionisation mass-spectrometry, MBC: Minimum bactericidal concentration, MIC : Minimum inhibitory concentration, MTT: Thiazolyl Blue Tetrazolium Bromide, MRSA: methicillin-resistant Staphylococcus aureus, NMR: nuclear magnetic resonance, TLC: thin layer chromatography, PD: prodelphinidin
  2,319 139 -
Antioxidant and hepatoprotective potential of phenol-rich fraction of Juniperus communis Linn. leaves
Akash Ved, Amresh Gupta, Ajay Kumar Singh Rawat
January-March 2017, 13(49):108-113
DOI:10.4103/0973-1296.197648  PMID:28216892
Background: Juniperus communis Linn. is an important plant in India traditional system of medicine which is widely used by different tribes in many countries. Objective: In the present study, the antioxidant, cytotoxic and hepatoprotective activities of Juniperus communis leaves were investigated against various models. Materials and Methods: ethanolic extract (70% v/v) of J. communis leaves was successively extracted using hexane and ethyl acetate to prepare various fractions. Total phenol content was resolute by the Folin-Ciocalteau's process. The antioxidant properties of the different fractions/extract of leaves of J. communis were examined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and Fe2+ chelating ability. Cytotoxic activity was examined by cell viability assay on HepG2 cells. Hepatoprotective activity of ethyl acetate fraction (EAF) evaluated against PCM-Paracetamol-induced hepatic damage in Wistar albino rats. Results: Total phenol content was found maximum 315.33 mg/GAE/g in EAF. Significant scavenging activity were found for EAF (IC50 = 177 μg/ml) as compared to standard BHT (IC50 = 138 μg/ml), while EAF showed good Fe2+ chelating ability having an IC50 value of 261 mg/ML compared to standard ethylenediaminetetraacetic acid (7.7 mg/mL). It was found that EAF treated group shows remarkable decrease in serum Aspartate aminotransferase, serum Alanine aminotransferase, total bilirubin, direct bilirubin, and alkaline phosphatase level in treatment group as compared to the hepatotoxic group. Conclusion: EAF of J. communis leaves is found to be potent antioxidant and hepatoprotective without any cytotoxicity and it can also be included in nutraceuticals with notable benefits for mankind or animal health. Abbreviations used: HepG2: Liver hepatocellular carcinoma, BHT: Butylated hydroxytoluene, PCM: Paracetamol, IC50: Half maximal inhibitory concentration, RSA: Radical Scavenging Activity, WST: Water-soluble tetrazolium.
  2,334 92 -
EDITORIAL
Pharmacognosy Magazine-Completion of 12 years of successful publishing
Mueen Ahmed
January-March 2017, 13(49):1-1
DOI:10.4103/0973-1296.197631  PMID:28216874
  2,304 102 -
ORIGINAL ARTICLES
Using UHPLC and UV-vis fingerprint method to evaluate substitutes for Swertia mileensis: An endangered medicinal plant
Jie Li, Ji Zhang, Hang Jin, Yuan-Zhong Wang, Heng-Yu Huang
January-March 2017, 13(49):13-20
DOI:10.4103/0973-1296.197655  PMID:28216877
Background: Millions of people are killed by viral hepatitis every year in the world, whereas many relevant medicines are too expensive to purchase. Swertia mileensis, a medicinal plant for hepatitis in the system of traditional Chinese medicine, has been vanishing gradually because of overexploitation. Objective: To find substitutes of S. mileensis and reduce the cost of purchasing drugs for hepatitis patients, the similarity of phytochemical constituents between S. mileensis and other three Swertia species was compared. Materials and Methods: Both ultra high performance liquid chromatographies and ultraviolet-vis fingerprints of four Swertia species were developed. Methanol extracts of the stems and leaves were used as samples to establish the fingerprint. The calibration curve was drawn for quantitative analysis of swertiamarin. The data of ultra high performance liquid chromatographies were evaluated statistically using similarity analysis and principal component analysis. Results: The result shows a significant difference at area of 204–290 nm in the ultraviolet fingerprint. Swertiamarin, the only one common peak, was defined in chromatographic fingerprints of four Swertia species. The quantitative analysis suggested that the highest concentration of swertiamarin is in S. davidii. The similarity indexes between different samples were almost under 0.60. In the principal component analysis, separate points not only represent the distinction among different species, but also perform chemical discrepancies in content between stems and leaves of one same species. Conclusions: S. angustifolia, S. davidii, and S. punicea are not suitable as substitutes of S. mileensis because of their remarkable differences in entirety and local part. In order to address issues about substitutes and high cost of purchasing drugs, more studies need to undertake. Abbreviation used: UHPLC: Ultra high performance liquid chromatography, UV-vis: Ultraviolet-vis, HBV: Anti-hepatitis virus, DNA: Deoxyribonucleic acid, PCA: Principal component analysis, D-GaIN: D-Galactosamine, BCG: Bacille Calmette-Guerin, LPS: Lipopolysaccharide
  2,274 94 -
Effects of potassium sulfate [K2SO4] on the element contents, polyphenol content, antioxidant and antimicrobial activities of milk thistle [Silybum marianum]
Gulsum Yaldiz
January-March 2017, 13(49):102-107
DOI:10.4103/0973-1296.197641  PMID:28216891
Background:Silybum marianum L. (Milk thistle) is native to the Mediterranean basin and is now widespread throughout the world. It's sprout is used as a herbal medicine for the treatment of liver disease for centuries. The seeds of milk thistle contain silymarin, an isomeric mixture of flavonolignans [silybin, silychristin, and silydianin. Silymarin acts as a strong anti-hepatotoxic. Objectives: The objective of this study was to evaluate the influences of potassium sulfate [K2SO4] fertilizer doses on polyphenol content, some nutrient elements, antioxidant and antimicrobial activities of milk thistle at experimental fields of Ordu University in Turkey. Methods: The antimicrobial activities of seed ethanol extracts and seed oil were tested in vitro against Pseudomonas aeruginosa (P. aeruginosa), Escherichia coli, (E. coli) Staphylococcus aureus (S. aureus), Aspergillus niger (A. niger) and Candida albicans (C. albicans) using the disc diffusion method. Free radical scavenging activity of the ethanolic extracts of milk thistle was determined spectrophotometrically by monitoring the disappearance of 2, 2-diphenyl-1-picrylhydrazil (DPPH·) at 517 nm according to the method described by Brand-Williams et al.[17] The phenolic contents in the ethanolic extracts of milk thistle were determined according to the procedure described by Slinkard and Singleton[19] with a slight modification of using a Folin-Ciocalteu phenolic reagent. The amount of total flavonoid in the ethanolic extracts was measured by aluminum chloride [AlCl3] colorimetric assay. The ions in aerosol samples were determined by using Dionex ICS 1100 Series ion chromatography. Results: Seed and seed oils obtained from obvious doses of potassium sulfate [0, 30, 60, 90 and 120 kg ha -1 fertilizer applications showed antimicrobial activities against E. coli, A. niger and P. aeruginosa. The application of 90 kg ha-1 of K2SO4 on seed oil resulted in the highest antimicrobial activities. At 100 µg mL-1and 200 µg mL-1, except the highest potassium application [120 kg ha -1 extract, all extracts showed high and similar DPPH scavenging activity. The highest phenolic compounds were obtained with 30 kg ha-1 of K2SO4, whereas the use of 60 kg ha-1 caused the highest total flavonoid content. This plant is a good source of K+, Ca+2, PO4-3, and Cl-1. Conclusion: In this study, increasing doses of potassium sulfate had significant effect on element, polyphenol content, antioxidant and antimicrobial activities of the milk thistle. Abbreviations used: AlCl3: aluminum chloride, Ca+2: calcium, Cl-: chloride, Cr: chromium CE: catechol equivalents, DPPH: 2,2-diphenylpicrylhydrazyl, ABTS: 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid, DAP: diamonyum fosfat, F-: fluoride, Fe: iron, K2SO4: potassium sulfate, K+ : potassium, Li+: lithium, Mg+2 : magnesium, NH4+ : amonyum, Na+: sodium, NO2-: nitrite, NO3-: nitrate, Ni: nickel, NaNO2: sodium nitrite, NaOH: sodium hidroksit. ND: Not detectable, PO4-3: phosphorus, Zn: zinc
  2,220 96 -
Rapid analysis of components in Coptis chinensis Franch by ultra-performance liquid chromatography with quadrupole time-of-flight mass spectrometry
Peng-peng Tian, Xiao-xu Zhang, Hong-ping Wang, Pu-ling Li, Yu-xin Liu, Shao-jing Li
January-March 2017, 13(49):175-179
DOI:10.4103/0973-1296.197635  PMID:28216903
Background: Coptis chinensis Franch is a traditional Chinese medical herb.Objective: In this article, ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was used to rapidly, qualitatively, and comprehensively identify the components in Coptis chinensis Franch. Materials and Methods: Chromatographic separation was achieved on an Agilent Zorbax RRHD Eclipse Plus C18 column. The mobile phase consisted of 0.1% formic acid water (A) and 0.1% formic acid acetonitrile (B) with a gradient program. Qualitative analysis was performed on an Agilent 6540 quadrupole time-of-flight mass spectrometer, which was equipped with a Dual AJS ESI source operating in negative mode. Results: A total of 30 alkaloid and non-alkaloid components of Coptis chinensis Franch were identified in only 14 min.Conclusion: This study helped to provide a basis for the quality control of Coptis chinensis Franch. Abbreviations used: Q-TOF-MS: quadrupole time-of-flight mass spectrometry, UPLC: ultra-performance liquid chromatography, pos: positive, neg: negative.
  2,167 143 -
Trichosanthes kirilowii exerts androgenic activity via regulation of PSA and KLK2 in 22Rv1 prostate cancer cells
Soo-Jin Jeong, Ji-Yoon Choi, Mi-Sook Dong, Chang-Seob Seo, Hyeun-Kyoo Shin
January-March 2017, 13(49):153-158
DOI:10.4103/0973-1296.197657  PMID:28216900
Background: The androgen comprises a group of hormones that play roles in male reproductive activity as well as personal characteristics. Objective: We investigated the androgenic activity of various herbal medicines in human prostate cancer 22Rv1 cells. Materials and Methods: Herbal extracts of Trichosanthes kirilowii (TK), Asarum sieboldii (AS), Sanguisorba officinalis (SO), and Xanthium strumarium (XS) were selected to have androgenic effects based on a preliminary in vitro screening system. Results: TK, AS, SO, and XS enhanced the proliferation of 22Rv1 cells without having cytotoxic effects. All tested herbal extracts increased androgen receptor (AR)-induced transcriptional activity in the absence or presence of dihydrotestosterone (DHT). In an AR-binding assay, TK, but not AS, SO, or XS, produced a significant inhibition of AR binding activity, indicating it has androgenic activity. Additionally, TK treatment positively regulated mRNA expression of the AR-related molecular targets prostate-specific antigen (PSA) and kallikrein 2 (KLK2) compared with untreated control. Conclusion: Taken together, TK-enhanced AR-mediated transcriptional activity might be an attractive candidate drug for treating androgen-related diseases. Abbreviations used: BPH: benign prostatic hyperplasia; AR: androgen receptor; DHT: dihydrotestosterone; PSA: prostate-specific antigen; TK: Trichosanthes kirilowii; AS: Asarum sieboldii; SO: Sanguisorba officinalis; XS: Xanthium strumarium; ATCC: American Type Culture Collection; FBS: fetal bovine serum; PBS: phosphate-buffered saline; SD: standard deviation; ARE: androgen-responsive element; KLK: kallikrein
  2,145 83 -
Evaluate the antigenotoxicity and anticancer role of β-sitosterol by determining oxidative DNA damage and the expression of phosphorylated mitogen-activated protein kinases', c-fos, c-jun, and endothelial growth factor receptor
Ramalingam Sharmila, Ganapathy Sindhu
January-March 2017, 13(49):95-101
DOI:10.4103/0973-1296.197634  PMID:28216890
Background: Plant sterols are the major source of micronutrients and have not shown any obvious side effects in human. β-sitosterol is one of the most prevalent phytosterols which have been recorded in ancient medicinal history for its use in the treatment of many chronic diseases,especially cancer. The modulations of mitogen-activated protein kinases' (MAPKs') play a crucial role in the development of human renal cell carcinoma. Objective: The aim of the current study is to evaluate the antigenotoxic and anticancer role of β-sitosterol against renal carcinogen. Materials and Methods: The extent of DNA damage was assessed by the comet assay. The status of p-p38 MAPK,p-c-Jun N-terminal kinase,p-extracellular-signal regulating kinase (ERK),c-fos,c-jun,and endothelial growth factor receptor (EGFR) were analyzed by western blot and polymerase chain reaction techniques. To further confirm the inhibition of ERK-2 by β-sitosterol,molecular docking study was performed. Results: Extensive DNA damage in acute study and a significant increase in levels of p-MAPKs',c-fos,c-jun,and EGFR was observed in N-diethylnitrosamine (200 mg/kg bw) and ferric nitrilotriacetate (9 mg/kg bw) alone treated rats. Rats which are pretreated with 20 mg/kg bw of β-sitosterol reduced the DNA damage and restored the elevated levels of above-mentioned markers (p < 0.05). The binding free energy obtained for β-sitosterol for ERK-2 was found to be-5.578. Conclusion: Therefore,it has been concluded that β-sitosterol has a strong potential against genotoxic as well as suppress neoplastic transformation in experimental renal cancer. Abbreviation used: AP-1: Activator protein-1,DEPC: Diethyl pyrocarbonate,EDTA: Ethylenediaminetetraacetic acid,EGFR: Endothelial growth factor receptor,ERK: Extracellular-signal regulating kinase,Fe-NTA: Ferric nitrilotriacetate,GAPDH: Glyceraldehyde-3-phosphate dehydrogenase,HBSS: Hank's balanced salt solution,JNK: c-Jun N-terminal kinase,MAPK: Mitogen-activated protein kinase,DEN: N-diethylnitrosamine,RCC: Renal cell carcinoma,SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis
  2,036 131 -
Simultaneous determination of eight phenolic acids, five saponins and four tanshinones for quality control of compound preparations containing Danshen-Sanqi herb-pair by HPLC-DAD
Hong Yao, Xiaomei Huang, Shaoguang Li, Youjia Wu, Xinhua Lin, Peiying Shi
January-March 2017, 13(49):64-75
DOI:10.4103/0973-1296.197651  PMID:28216885
Background: The herb-pair, Salviaemiltiorrhizae (Danshen, DS) and Panaxnotoginseng (Sanqi,SQ), often occurs in traditional Chinese medicine prescriptions used for the treatment of cardiovascular diseases in clinics in Asian areas. Many commercial preparations containing the DS-SQ herb-pair were produced by various manufactures with the different production process. The raw materials were from different sources, which raised a challenge to control the quality of the herb-pair medicines. Objective: In this paper, a high-performance liquid chromatography (HPLC) method was developed to simultaneously determine seventeen bioactive components, including 8 phenolic acids, 4 tanshinones, and 5 saponins, for quality control of compound preparations containing DS-SQ herb-pair. The chromatographic separation was studied on an UltimateTM XB-C18 column (150 mm × 4.6 mmi.d., 3.5 μm) with a mobile phase composed of 0.5% aqueous acetic acid and acetonitrile using a gradient elution in 70 min. Results: The optimum detection wavelength was set at 288 nm for phenolic acids and tanshinones, and 203 nm for saponins. The method was validated sufficiently by examining the precision, recoveries, linearity, range, LOD and LOQ, and was successfully applied to quantify the seventeen compounds in five commercial preparations containing DS-SQ herb-pair. Conclusions: It is the first time to report the rapid and simultaneous analysis of the seventeen compounds with the base-line separation of peaks for ginsenoside Rg1 and Re in 70 min by routine HPLC. This HPLC method could be considered as good quality criteria to control the quality of preparations containing DS-SQ herb-pair. Abbreviations used: DS: Salviae miltiorrhizae; SQ: Panaxnotoginseng; HPLC: high-performance liquid chromatography; DAD: diode array detector; LOD: limit of detection; LOQ: limit of quantification; TCMs: Traditional Chinese medicines; GDDP: Guanxin Danshen dripping pills; FDDP: Fufang Danshen dripping pills; FDT: Fufang Danshen tablets; FDC: Fufang Danshen capsules; GP: Guanxin pills
  2,054 104 -
Chemical components from Aloe and their inhibition of indoleamine 2, 3-dioxygenase
Ya Nan Sun, Lin Ying Li, Wei Li, Jong Seong Kang, Inkyu Hwang, Young Ho Kim
January-March 2017, 13(49):58-63
DOI:10.4103/0973-1296.197633  PMID:28216884
Background: In Korea, Aloe is routinely ingested as a traditional medicine or as a component of health beverages. Objective: To research the inhibition of indoleamine 2, 3-dioxygenase (IDO) activities of components from Aloe. Materials and Methods: the compounds were isolated by a combination of silica gel and YMC Rp-18 column chromatography, and their structures were identified by analysis of spectroscopic data (1D, 2D-NMR, and MS). All of the isolated compounds were examined for their ability to inhibit IDO, which actively suppresses immune functions by catalyzing the rate limiting reaction in the conversion of tryptophan to kynurenine. Results: In this phytochemical study, 18 known compounds were isolated from aqueous dissolved Aloe exudates. All of the isolated compounds were examined for their ability to inhibit IDO activities for a series of anthraquinone derivatives (1-7) isolated from the Aloe extract; the IC50 values of these compounds ranged from 39.41 to 53.93 µM. Enzyme kinetic studies of their modes of inhibition indicated that all of the compounds were uncompetitive inhibitors. Conclusion: The aqueous dissolved Aloe exudate can be used as a source of novel natural IDO inhibitors and merit testing as therapeutic agents in the treatments of cancer and immunopathologic diseases, such as autoimmune, inflammatory, and allergic disorders. Abbreviation used: IDO: inhibit indoleamine 2, 3-dioxygenase, TMS: tetramethylsilane, HMQC: heteronuclear multiple quantum correlation, HMBC: heteronuclear multiple bond correlation, COSY: 1H-1H correlation spectroscopy, ESI-MS: Electrospray ionization mass spectrometry, DMSO: dimethyl sulfoxide
  2,037 100 -
Osteoblasts proliferation and differentiation stimulating activities of the main components of Epimedii folium
Mingming Liu, Haiyan Xu, Yong Ma, Jian Cheng, Zhen Hua, Guicheng Huang
January-March 2017, 13(49):90-94
DOI:10.4103/0973-1296.197654  PMID:28216889
Background: Osteoporosis is a disease of bones that leads to an increased risk of fracture. Epimedii Folium is commonly used for treating bone fractures and joint diseases for thousands of years in China. Methods: This study was aimed to screen active components, which might have the potency to stimulate osteoblasts proliferation and differentiation in Epimedii Folium. An HPLC method was established to analyze the main components in Epimedii Folium. The MTT and ALP methods were utilized for the assay of osteoblasts proliferation and differentiation activity. Bavachin, a flavonoid compound was treated as the positive control. Results: Totally eight compounds have been identified by comparing their retention time with correspondent standard substances. Icariside I and icariside II significantly stimulated cell proliferation and osteoblasts differentiation. All these compounds were found with a characterized flavonoid structure in each of their molecule backbones. Conclusion: These results lead to a hypothesis that flavonoid monoglycoside structure might be crucial to exhibit the activity. The structure–effect relationship of these compounds with flavonoid monoglycoside structure in mouse primary calvarial osteoblasts needs to be explored in further research. Abbreviations used: HPLC: High performance liquid chromatography, MTT: Methylthiazolyldiphenyl - tetrazolium bromide, ALP: Alkaline phosphatase
  1,989 144 -
Therapeutic effect of Dendrobium candidum on lupus nephritis in mice
Qiang Wang, Peng Sun, Rui Wang, Xin Zhao
January-March 2017, 13(49):129-135
DOI:10.4103/0973-1296.197653  PMID:28216896
Context: Dendrobium candidum (D.candimum) widely is a functional drug. The curative effect of D. candidum on lupus nephritis has been studied in vivo. Materials and Method: The DBA/2 and B6D2F1 mice were used for this in vivo experiment. The 50% effective dose (ED50) was used to check the effective concentration for this study. Then the SCr, BUN, TC, TG, IL-6, IL-12, TNF-α, and IFN-γ levels were determined by kits. The output of urine protein was determined by means of Coomassie Brilliant Blue, and the auto-antibody dsDNA was determined with titer plate technology and indirect immunofluorescence. The NF-κB, IκB-α, TGF 'β1, Fas, and FasL expressions were measured by RT-PCR and western blot assay. The component analysis of D. candidum was determined by nuclear magnetic resonance. Results: Based on the ED50 result at 329 mg/kg, 200 and 400 mg/kg doses were chosen for this study. SCr, BUN, TC and TG levels of 400 mg/kg D. candidum mice were lower than control mice, TP and ALB levels were higher than control mice. The control and 400 mg/kg treated mice tested positive for dsDNA at the end of sixth and tenth week after the experiment began. The glomerular number of 400 mg/kg treated mice was more than control group. Treatment with 400 mg/kg D. candidum reduced IL-6, IL-12, TNF-α and IFN-γcytokine levels as compared to control mice. D. candidum decreased NF-κb, TGF 'β1, Fas, FasL and increased IκB-α expressions in kidney tissue. There were 11 compounds in dry D. candidum, these compounds might make the curative effects of lupus nephritis. Conclusion: D. candidum showed a potential curative effect on lupus nephritis. It could be used as a health medicine on lupus nephritis. Abbreviations used: LN: Lupus nephritis, SLE: systemic lupus erythematosus, D. candidum: Dendrobium candidum; IL-6: interleukin-6, IL-12: interleukin-12, TNF-α: tumor necrosis factor alpha, IFN-γ: Interferon-gamma, SCr: serum creatinine, BUN: blood urea nitrogen, TC: total cholesterol, TG: triglyceride, TP: total protein, ALB: albumin.
  2,008 86 -
Specific PCR identification between Peucedanum praeruptorum and Angelica decursiva and identification between them and adulterant using DNA barcode
Bang-Xing Han, Yuan Yuan, Lu-Qi Huang, Qun Zhao, Ling-Ling Tan, Xiang-Wen Song, Xiao-Mei He, Tao Xu, Feng Liu, Jian Wang
January-March 2017, 13(49):38-45
DOI:10.4103/0973-1296.197658  PMID:28216881
Background: The traditional Chinese medicine (TCM) Qianhu and Zihuaqianhu are the dried roots of Peucedanum praeruptorum and Angelica decursiva, respectively. Since the plant sources of Qianhu and Zihuaqianhu are more complex, the chemical compositions of P. praeruptorum andA. decursiva are significantly different, and many adulterants exist because of the differences in traditional understanding and medication habits. Therefore, the rapid and accurate identification methods are required. Objective: The aim was to study the feasibility of using DNA barcoding to distinguish between Traditional Chinese medicine Qianhu (Peucedanum praeruptorum), Zihuaqianhu (Angelica decursiva), and common adulterants, based on internal transcribed spacer (ITS) sequences, as well as specific PCR identification between P. praeruptorum and A. decursiva. Materials and Methods: The ITS sequences of P. praeruptorum, A. decursiva, and adulterant were studied, and a phylogenetic tree was constructed. Based on the ITS barcode, the specific PCR primer pairs QH-CP19s/QH-CP19a and ZHQH-CP3s/ZHQH-CP3a were designed for P. praeruptorum and A. decursiva, respectively. The amplification conditions were optimized, and specific PCR products were obtained. Results: The results showed that the phylogenetic trees constructed using the BI and MP methods were consistent, and P. praeruptorum and A. decursiva sequence haplotypes formed their own monophyly. The experimental results showed that in PCR products, the target bands appeared in the genuine drug and not in the adulterant, which suggests the high specificity of the two primer pairs. Conclusion: The ITS sequence was ideal DNA barcode to identify P. praeruptorum, A. decursiva, and adulterant. The specific PCR is a quick and effective method to distinguish between P. praeruptorum and A. decursiva. Abbreviations used: TCM: The traditional Chinese medicine, P.: Peucedanum, A.: Angelica, ITS: The internal transcribed spacer, PCR: Polymerase chain reaction, NCBI: National Center for Biotechnology Information, NI: Number of individuals, HN: Haplotype number; GAN: Gen Bank accession numbers, L.: Ligusticum, O.: Ostericum, A.: Angelica, P.: Pimpinella, BI: Bayesian inference, MP: Maximum parsimony, AIC: Akaike Information Criterion, MCMC: Markov Chains Monte Carlo, TBR: Tree bisection-reconnection, LPP: Length of PCR product, PRP: PCR reaction procedure, SNP: Single nucleotide polymorphisms, PP: Posterior probability, BS: Bootstrap.Qun Zhao
  1,950 93 -
Combinative method using multi-components quantitation and HPLC fingerprint for comprehensive evaluation of Gentiana crassicaulis
Jiuhua Song, Fengzheng Chen, Jiang Liu, Yuanfeng Zou, Yun Luo, Xiaoyan Yi, Jie Meng, Xingfu Chen
January-March 2017, 13(49):180-187
DOI:10.4103/0973-1296.197639  PMID:28216904
Background: Gentiana crassicaulis () is an important traditional Chinese herb. Like other herbs, its chemical compounds vary greatly by the environmental and genetic factors, as a result, the quality is always different even from the same region, and therefore, the quality evaluation is necessary for its safety and effective use. In this study, a comprehensive method including HPLC quantitative analysis and fingerprints was developed to evaluate the quality of Cujingqinjiao and to classify the samples collected from Lijiang City of Yunnan province. A total of 30 common peaks including four identified peaks, were found, and were involved for further characterization and quality control of Cujingqinjiao. Twenty-one batches of samples from Lijiang City of Yunnan Province were evaluated by similarity analysis (SA), hierarchical cluster analysis (HCA), principal component analysis (PCA) and factor analysis (FA) according to the characteristic of common peaks.Results: The obtained data showed good stability and repeatability of the chromatographic fingerprint, similarity values were all more than 0.90. This study demonstrated that a combination of the chromatographic quantitative analysis and fingerprint offered an efficient way to quality consistency evaluation of Cujingqinjiao. Consistent results were obtained to show that samples from a same origin could be successfully classified into two groups.Conclusion: This study revealed that the combinative method was reliable, simple and sensitive for fingerprint analysis, moreover, for quality control and pattern recognition of Cujingqinjiao. Abbreviations used: SA: Similarity analysis, HCA: Hierarchical cluster analysis, PCA :Principal component Analysis, FA :Factor analysis
  1,918 89 -
Comparative pharmacokinetic profiles of three protoberberine-type alkaloids from raw and bile-processed Rhizoma coptidis in heat syndrome rats
Yuan Zi-min, Chen Yue, Gao Hui, Lv Jia, Chen Gui-rong, Jing Wang
January-March 2017, 13(49):51-57
DOI:10.4103/0973-1296.197632  PMID:28216883
Background: The Bile-processed Rhizoma coptidis (BRC), which has a colder drug property than Rhizoma coptidis (RC), is widely used for the treatment of heat syndrome. We compared the pharmacokinetics of the protoberberine-type alkaloids in BRC and RC in rats with heat syndrome to elucidate the bile-processing mechanism. Material and Methods: We established a rapid and sensitive method for simultaneously determining three alkaloids: berberine, palmatine, and jatrorrhizine, in rat plasma based on ultra-performance liquid chromatography/tandem mass spectrometry. The separation was carried out on a Waters ACQUITY BEA C18 column. The mobile phase consisted of acetonitrile (containing 0.1% formic acid) and water (containing 0.1% formic acid and 10 mmol/L ammonium acetate) and carbamazepine was used as an internal standard. The detection was carried out in a multiple reaction monitoring mode (MRM) using electrospray ionization in the positive ion mode. Results: Pharmacokinetic profiles indicated that the Cmax of berberine and palmatine increased two times and the Tmax of the three alkaloids decreased three times after bile processing. AUC0→∞ and AUC0→t of the alkaloids were similar between RC and BRC. Conclusion: The results suggest that bile processing could increase the absorption rate of alkaloids. This study broadens our understanding of Chinese herbal medicine processing. Abbreviation Used: RC: Rhizoma coptidis, BRC: Bile-processed Rhizoma coptidis, HPLC : high-performance liquid chromatography, UPLC-MS/MS: ultra-performance liquid chromatography-mass spectrometry/ mass spectrometry, LC-MS: liquid chromatography-mass spectrometry, MRM : multiple reaction monitoring mode, QC: quality control, RE: relative error, RSD: relative standard deviation, Cmax: maxium of drug concentration, Tmax: time for maxium of drug concentration, AUC: area under concentration-time curve, LLOQ: Linearity and lower limits of quantification, t1/2: half-life, Clz: body clearance
  1,891 105 -
Obtaining a dry extract from the Mikania laevigata leaves with potential for antiulcer activity
Mariana Viana Pinto, Ezequiane Machado Oliveira, Jose Luiz Rodrigues Martins, Jose Realino de Paula, Elson Alves Costa, Edemilson Cardoso da Conceição, Maria Teresa Freitas Bara
January-March 2017, 13(49):76-80
DOI:10.4103/0973-1296.197640  PMID:28216886
Background: Mikania laevigata leaves are commonly used in Brazil as a medicinal plant.Objective: To obtain hydroalcoholic dried extract by nebulization and evaluate its antiulcerogenic potential.Materials and Methods: Plant material and hydroalcoholic extract were processed and analyzed for their physicochemical characteristics. A method using HPLC was validated to quantify coumarin and o-coumaric acid. Hydroalcoholic extract was spray dried and the powder obtained was characterized in terms of its physicochemical parameters and potential for antiulcerogenic activity.Results: The analytical method proved to be selective, linear, precise, accurate, sensitive, and robust. M. laevigata spray dried extract was obtained using colloidal silicon dioxide as adjuvant and was shown to possess 1.83 ± 0.004&#37; coumarin and 0.80 ± 0.012% o-coumaric acid. It showed significant antiulcer activity in a model of an indomethacin-induced gastric lesion in mice and also produced a gastroprotective effect.Conclusion: This dried extract from M. laevigata could be a promising intermediate phytopharmaceutical product. Abbreviations used: DE: M. laevigata spray dried extract, HE: hydroalcoholic extract
  1,912 78 -
Feeble antipyretic, analgesic, and anti-inflammatory activities were found with regular dose 4'-O-β-D-glucosyl-5-O-methylvisamminol, one of the conventional marker compounds for quality evaluation of Radix Saposhnikoviae
Jing-Ming Yang, Hua Jiang, Hong-Liang Dai, Zi-Wei Wang, Gui-Zhi Jia, Xiang-Cai Meng
January-March 2017, 13(49):168-174
DOI:10.4103/0973-1296.197637  PMID:28216902
Introduction: 4'-O-β-D-glucosyl-5-O-methylvisamminol (GML) is a conventional marker compound for quality control of Radix Saposhnikoviae. Despite that, neither pharmacodynamic or pharmacokinetic information is available with regard to GML. As such, the aim of thisstudy was to assess the conventional evaluation indices for the quality of Radix Saposhnikoviae. Materials and methods: Pyretic animal model, hot plate test, and ear edema model were established to evaluate and compare the antipyretic, analgesic, and anti-inflammatory effect of the chromone derivativescimifugin, prime-O-glucosylcimifugin (PGCN), and GML in Radix Saposhnikoviae. High performance liquid chromatography separation and analysis was used to obtain pharmacokinetic parameters. Simulated gastric fluid and simulated intestinal fluid was used to investigate the metabolite profiles of PGCN and GML in gastrointestinal tract. Results: Cimifugin exerted a marked dose-dependent antipyretic, analgesic, and anti-inflammatory effect,whereas the effects of PGCN were relatively lower. GML had feeble pharmacodynamic effects. Pharmacokinetic study showed that only cimifugin was detected in the plasma sample of cimifugin and PGCN-treated animals, with drug concentration in the former much higher than the latter. No components were traced in the plasma samples from GML-treated rats. Stability study showed that PGCN and GML was predominantly biotransformed into cimifugin and 5-O-methyvisammiol, respectively. The latter was proven to be extremely unstable in liver tissue homogenate and plasma.Conclusions: A feeble antipyretic, analgesic, and anti-inflammatory activities was observed when GML was orally delivered. Given that Radix Saposhnikoviae extract is generally administered orally, we speculate that this compound might be a nonpharmacolagically active agent in real usage. Thus, it might be unscientific to evaluate the quality of Radix Saposhnikoviae based on the content of GML. Abbreviations used: AUC:area under concentration-time curve, DNP:2,4-Dinitrophenol, HPLC:high performance liquid chromatography, HPLC-MS:high performance liquid chromatography- mass spectrography, GML:4'-O-β-D-glucosyl-5-O-methylvisamminol, MVL:5-O-methyvisammiol, PGCN:prime-O-glucosylcimifugin, SGF:alkaline phosphatase. SIF:simulated intestinal fluid
  1,769 94 -
High performance liquid chromatography-diode array detector method for the simultaneous determination of five compounds in the pulp and seed of sea buckthorn
Lu Zhao, E Wen, Halmuart Upur, Shuge Tian
January-March 2017, 13(49):136-140
DOI:10.4103/0973-1296.197656  PMID:28216897
Context: Sea buckthorn (Hippophae rhamnoides L.) as a traditional Chinese medicinal plant has various uses in Xinjiang. Objective: A reversed-phase rapid-resolution liquid-chromatography method with diode array detector was developed for simultaneous determination of protocatechuic acid, rutin, quercetin, kaempferol, and isorhamnetin in the pulp and seed of sea buckthorn, a widely used traditional Chinese medicine for promoting metabolism and treating scurvy and other diseases. Settings and design: Compounds were separated on an Agilent ZORBAX SB-C18 column (4.6 mm × 250 mm, 5 μm; USA) with gradient elution using methanol and 0.4% phosphoric acid (v/v) at 1.0 mL/min. Detection wavelength was set at 280 nm. Materials and Methods: The fruits of wild sea buckthorn were collected from Wushi County in Aksu, Xinjiang Province. Statistical performances: The RSD of precision test of the five compounds were in the range of 0.60-2.22%, and the average recoveries ranged from 97.36% to 101.19%. Good linearity between specific chromatographic peak and component qualities were observed in the investigated ranges for all the analytes (R2 > 0.9997). Results: The proposed method was successfully applied to determine the levels of five active components in sea buckthorn samples from Aksu in Xinjiang. Conclusions: The proposed method is simple, fast, sensitive, accurate, and suitable for quantitative assessment of the pulp and seed of sea buckthorn. Abbreviation used: PR: protocatechuic acid, RU: rutin, QU: quercetin, KA: kaempferol, IS: isorhamnetin, HPLC: high-performance liquid chromatography, HPLC-DAD: high performance liquid chromatographydiode array detector, LOD: linearity and limit of detection, LOQ: limit of quantitation, RSD: relative standard deviation
  1,724 109 -
Effects of insecticidal ketones present in mint plants on GABAA receptor from mammalian neurons
Mariela Eugenia Sánchez-Borzone, Leticia Delgado Marin, Daniel Asmed García
January-March 2017, 13(49):114-117
DOI:10.4103/0973-1296.197638  PMID:28216893
Background: The genus Mentha, an important member of the Lamiaceae family, is represented by many species commonly known as mint. The insecticidal activity of Mentha oil and its main components has been tested and established against various insects/pests. Among these, the ketone monoterpenes that are most common in different Mentha species demonstrated insect toxicity, with pulegone being the most active, followed by carvone and menthone. Considering that the GABAA receptor (GABAA-R) is one of the main insecticide targets on neurons, and that pulegone would modulate the insect GABA system, it may be expected that the insecticidal properties of Mentha ketones are mediated by their interaction with this receptor. Objective: In order to discern the pharmacological actions of these products when used as insecticides on mammalian organisms, we evaluated the pharmacologic activity of ketones, commonly present in Mentha plants, on native GABAA-R from rats. Materials and Methods: Determination of ketones effects on allosterically enhanced benzodiazepine binding, using primary cultures of cortical neurons, which express functional receptors and MTT assay to evaluate their cell toxicity. Results: Our results seem to indicate that ketone components of Mentha, with proven repellent or insecticide activity, were able to behave as GABAA-R negative allosteric modulators in murine cells and consequently could exhibit convulsant activity in mammalians. Only pulegone at the highest assayed concentration (2 mM) showed a significant reduction in cell viability after exposure for 24 hr. Conclusion: The present results strongly suggest that the ketone components of Mentha are able to exhibit convulsant activity in mammalian organisms, but functional assays and β€͸in vivo experiments would be necessary to corroborate this proposed action. Abbreviations used: GABA: gamma aminobutyric acid, GABAA-R: GABAA receptor, MTT: 1-(4,5-dimethylthiazol-2-yl)-3,5-diphenylformazam, DMEM: Dulbecco's modified minimum essential mèdium, [3H]TBOB: [3H] t-Butylbicycloorthobenzoate
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