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   2015| April-June  | Volume 11 | Issue 42  
    Online since March 12, 2015

 
 
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ORIGINAL ARTICLES
Quercetin induces human colon cancer cells apoptosis by inhibiting the nuclear factor-kappa B Pathway
Xiang-An Zhang, Shuangxi Zhang, Qing Yin, Jing Zhang
April-June 2015, 11(42):404-409
DOI:10.4103/0973-1296.153096  PMID:25829782
Quercetin can inhibit the growth of cancer cells with the ability to act as chemopreventers. Its cancer-preventive effect has been attributed to various mechanisms, including the induction of cell-cycle arrest and/or apoptosis as well as the antioxidant functions. Nuclear factor kappa-B (NF-kB) is a signaling pathway that controls transcriptional activation of genes important for tight regulation of many cellular processes and is aberrantly expressed in many types of cancer. Inhibitors of NF-kB pathway have shown potential anti-tumor activities. However, it is not fully elucidated in colon cancer. In this study, we demonstrate that quercetin induces apoptosis in human colon cancer CACO-2 and SW-620 cells through inhibiting NF-kB pathway, as well as down-regulation of B-cell lymphoma 2 and up-regulation of Bax, thus providing basis for clinical application of quercetin in colon cancer cases.
  11 3,670 15
In silico analysis and molecular docking studies of potential angiotensin-converting enzyme inhibitor using quercetin glycosides
Syed Aun Muhammad, Nighat Fatima
May 2015, 11(42):123-126
DOI:10.4103/0973-1296.157712  PMID:26109757
The purpose of this study was to analyze the inhibitory action of quercetin glycosides by computational docking studies. For this, natural metabolite quercetin glycosides isolated from buckwheat and onions were used as ligand for molecular interaction. The crystallographic structure of molecular target angiotensin-converting enzyme (ACE) (peptidyl-dipeptidase A) was obtained from PDB database (PDB ID: 1O86). Enalapril, a well-known brand of ACE inhibitor was taken as the standard for comparative analysis. Computational docking analysis was performed using PyRx, AutoDock Vina option based on scoring functions. The quercetin showed optimum binding affinity with a molecular target (angiotensin-converting-enzyme) with the binding energy of −8.5 kcal/mol as compared to the standard (−7.0 kcal/mol). These results indicated that quercetin glycosides could be one of the potential ligands to treat hypertension, myocardial infarction, and congestive heart failure.
  5 2,746 18
Sesamol, a lipid lowering agent, ameliorates aluminium chloride induced behavioral and biochemical alterations in rats
Jessy John, Madhavan Nampoothiri, Nitesh Kumar, Jayesh Mudgal, Gopalan Kutty Nampurath, Mallikarjuna Rao Chamallamudi
April-June 2015, 11(42):327-336
DOI:10.4103/0973-1296.153086  PMID:25829772
Background: Sesame oil from the seeds of Sesamum indicum Linn. (Pedaliaceae) has been used traditionally in Indian medical practice of Ayurveda in the treatment of central nervous system disorders and insomnia. A few published reports favor the anti-dementia effect of sesamol (SML), an active constituent of sesame oil. Objective: Thus, the present study was aimed to explore the anti-dementia effect and possible mechanism (s) of SML in aluminium chloride (AlCl 3 )-induced cognitive dysfunction model in rodents with special emphasis on memory centers viz., hippocampus and frontal cortex. Methods: Male Wistar rats were exposed to AlCl 3 (175 mg/kg p.o.) for 60 days. SML (10 and 20 mg/kg) and rivastigmine (1 mg/kg) were administered orally 45 min before administration of AlCl 3 for 60 days. Spatial memory was assessed using Morris water maze test. After 60 days of treatment animals were sacrificed, hippocampus and frontal cortex were collected and analyzed for acetylcholinesterase (AChE) activity, tumor necrosis factor (TNF-α) level, antioxidant enzymes (Glutathione, catalase), lipid peroxidation, and nitrite level. The circulating triglycerides, total cholesterol, low-density lipoprotein (LDL) and high-density lipoprotein (HDL) levels were also analyzed. Results: SML significantly prevented behavioral impairments in aluminium-exposed rats. Treatment with SML reversed the increased cholesterol, triglycerides and LDL while raised the HDL levels. SML significantly corrected the effect of AlCl 3 on AChE activity. Further, SML reversed the elevated nitric oxide, TNF-α and reduced antioxidant enzymes in hippocampus and frontal cortex. Conclusion: The present study suggests the neuro-protection by SML against cognitive dysfunction induced by environmental toxin (AlCl 3 ) in hippocampus and frontal cortex.
  5 3,305 18
Shikonin induces apoptosis in the human gastric cancer cells HGC-27 through mitochondria-mediated pathway
Yue Hou, Jinghua Xu, Xia Liu, Xichun Xia, Ning Li, Xiuli Bi
April-June 2015, 11(42):250-256
DOI:10.4103/0973-1296.153074  PMID:25829762
Background: Gastric cancer (GC) is one of the most frequently occurring digestive tract cancers and fewer chemotherapeutic drugs for GC have shown promising results. In this study, we investigated the anti-tumor activity of shikonin, a natural compound isolated from the Chinese plant Lithospermum erythrorhizon, against the human GC cell line HGC-27. Materials and Methods: HGC-27 cells treated with shikonin at a concentration of 30μM or above showed significant growth inhibition compared to control cells. Shikonin-treated cells also underwent apoptosis as detected by flow cytometric analysis and microscopic examination of cellular morphology. Further investigation into the underlying mechanism of apoptosis by western blot showed that the shikonin promoted the activation of poly-(ADP-ribose)-polymerase, caspase-3 and caspase-9 following 24 h or 48 h of treatment time, as well as the activation of caspase-8, but only after 48 h of treatment time. Furthermore, the levels of mitochondrial membrane potential, B-cell lymphoma 2 (Bcl-2) and Bcl-extra large were reduced following shikonin treatment while the level of Bax was increased. In addition, shikonin also caused a signifi cant reduction of the protein Survivin, while having little effect on the expression on X-linked inhibitor of apoptosis protein. Conclusion: Taken together, these results showed that the shikonin exhibited its anti-tumor activity against HGC-27 cells through inhibiting cell growth and promoting apoptosis by targeting mitochondrial-related signaling pathway. Our finding may represent a positive step in finding a natural and effective compound that could be important implication for future development of chemotherapeutic and/or chemopreventive agent against GC.
  4 3,282 14
Methyl gallate isolated from Spondias pinnata exhibits anticancer activity against human glioblastoma by induction of apoptosis and sustained extracellular signal-regulated kinase 1/2 activation
Dipankar Chaudhuri, Nikhil Baban Ghate, Sudhir Shankar Singh, Nripendranath Mandal
April-June 2015, 11(42):269-276
DOI:10.4103/0973-1296.153078  PMID:25829764
Background: Spondias pinnata has been reported for its efficient anticancer effects, but the studies were mostly focused on its extract. Objective: Since its bioactive compounds are largely unknown, this study was designed to characterize the lead components present in it and their anticancer activity against human glioblastoma cell line (U87). Materials and Methods: Major compounds from the ethyl acetate fraction were isolated by column chromatography and their anticancer potentials against U87 cells were evaluated. Furthermore, flow cytometric and immunoblotting analyses were performed to demonstrate the mechanism of apoptosis inducing activity of methyl gallate (MG) against U87 cell line. Results: Four major compounds were isolated from the ethyl acetate fraction. Amongst these, two compounds showed promising activities and with the help of different spectroscopic methods they were identified as gallic acid and MG. Flow cytometric studies revealed that MG-induced apoptosis in U87 cells dose-dependently; the same was confirmed by activation of caspases through cleavage of endogenous substrate poly (adenosine diphosphate-ribose) polymerase. MG treatment also induced the expression of p53 and B-cell lymphoma-2-associated X and cleavage of BH3 interacting-domain with a concomitant decrease in B-cell lymphoma-2 expression. Moreover, MG-induced sustained phosphorylation of extracellular signal-regulated kinase (ERK1/2) in U87 cells with no change in the phosphorylation of other mitogen-activated protein kinases (c-Jun N-terminal of stress-activated protein kinases, p38). Conclusion: MG is a potent antioxidant and it induces sustained ERK1/2 activation and apoptosis in human glioblastoma U87, and provide a rationale for evaluation of MG for other brain carcinoma cell lines for the advancement of glioblastoma therapy.
  4 2,846 14
The potential of selected Australian medicinal plants with anti-Proteus activity for the treatment and prevention of rheumatoid arthritis
IE Cock, V Winnett, J Sirdaarta, B Matthews
May 2015, 11(42):190-208
DOI:10.4103/0973-1296.157734  PMID:26109767
Background: A wide variety of herbal medicines are used in indigenous Australian traditional medicinal systems to treat rheumatoid arthritis (RA) and inflammation. The current study was undertaken to test the ability of a panel of Australian plants with a history of the ethnobotanical usage in the treatment of inflammation for the ability to block the microbial trigger of RA. Materials and Methods: One hundred and six extracts from 40 plant species were investigated for the ability to inhibit the growth of the bacterial trigger of RA (Proteus mirabilis). The extracts were tested for toxicity in the Artemia nauplii bioassay. The most potent inhibitor of P. mirabilis growth was further analyzed by reversed-phase high performance liquid chromatography (RP-HPLC) coupled to high accuracy time-of-flight (TOF) mass spectroscopy. Results: Sixty-five of the 106 extracts tested (61.3%) inhibited the growth of P. The Aleurites moluccanus, Datura leichardtii, Eucalyptus major, Leptospermum bracteata, L. juniperium, Macadamia integriflora nut, Melaleuca alternifolia, Melaleuca quinquenervia, Petalostigma pubescens, P. triloculorae, P. augustifolium, Scaevola spinescens, Syzygium australe, and Tasmannia lanceolata extracts were determined to be the most effective inhibitors of P. mirabilis growth, with minimum inhibitory concentration (MIC) values generally significantly below 1000 μg/ml. T. lanceolata fruit extracts were the most effective P. mirabilis growth inhibitors, with a MIC values of 11 and 126 μg/ml for the methanolic and aqueous extracts, respectively. Subsequent analysis of the T. lanceolata fruit extracts by RP-HPLC coupled to high-resolution TOF mass spectroscopy failed to detect resveratrol in either T. lanceolata fruit extract. However, the resveratrol glycoside piceid and 2 combretastatin stilbenes (A-1 and A-4) were detected in both T. lanceolata fruit extracts. With the exception of the Eucalyptus and Syzygium extracts, all extracts exhibiting Proteus inhibitory activity were also shown to be nontoxic, or of low toxicity in the Artemia nauplii bioassay. Conclusions: The low toxicity of these extracts and their inhibitory bioactivity against Proteus spp. indicate their potential in blocking the onset of rheumatoid arthritis.
  4 2,451 15
Cytotoxicity of some edible mushrooms extracts over liver hepatocellular carcinoma cells in conjunction with their antioxidant and antibacterial properties
Gökhan Sadi, Bugrahan Emsen, Abdullah Kaya, Aytaç Kocabas, Seval Çinar, Deniz Irtem Kartal
May 2015, 11(42):6-18
DOI:10.4103/0973-1296.157665  PMID:26109775
Background: Mushrooms have been valued for their nutritive content and as traditional medicines; several important medicinal properties of mushrooms have been recognized worldwide. Objective: The purpose of this study was to elucidate the cell growth inhibitory potential of four edible mushrooms; Coprinus comatus (O.F. Mull.) Pers. (Agaricaceae), Tricholoma fracticum (Britzelm.) Kreisel (Tricholomataceae), Rhizopogon luteolus Fr. and Nordholm (Rhizopogonaceae), Lentinus tigrinus (Bull.) Fr. (Polyporaceae) on hepatocellular carcinoma (HepG2) cells in conjunction with their antioxidant and antibacterial capacities. Materials and Methods: Five different extracts of edible mushrooms were obtained using water, methanol, acetone, n-hexane and chloroform as solvent systems for cytotoxic, antioxidant and antibacterial properties. Results: C. comatus showed substantial in vitro cytotoxic activity against HepG2 cell lines with all extracts especially with chloroform 50% inhibition (IC 50 value of 0.086 mg/ml) and acetone (IC 50 value of 0.420 mg/ml). Chloroform extract of C. comatus had maximum amount of β-carotene (25.94 μg/mg), total phenolic content (76.32 μg/mg) and lycopene (12.00 μg/mg), and n-hexane extract of L. tigrinus had maximum amount of flavonoid (3.67 μg/mg). While chloroform extract of C. comatus showed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) capturing activity (1.579 mg/ml), the best result for metal chelating activity was obtained from methanolic extract (0.842 mg/ml). Moreover, all tested mushrooms demonstrated antibacterial activity and n-hexane extract of L. tigrinus and acetone extracts of T. fracticum were the most active against tested microorganism. Conclusion: These results indicate that different extracts of investigated mushroom have considerable cytotoxic, antioxidant and antibacterial properties and may be utilized as a promising source of therapeutics.
  4 2,688 16
Experimental treatment of breast cancer-bearing BALB/c mice by artemisinin and transferrin-loaded magnetic nanoliposomes
Amir Gharib, Zohreh Faezizadeh, Seyed Ali Reza Mesbah-Namin, Ramin Saravani
May 2015, 11(42):117-122
DOI:10.4103/0973-1296.157710  PMID:26109756
Background: The combination of artemisinin and transferrin exhibits versatile anticancer activities. In previous, we successfully prepared artemisinin and transferrin-loaded magnetic nanoliposomes and evaluated their anti-proliferative activity against MCF-7 and MDA-MB-231 cell lines in vitro. In this study, we investigate the in vivo anti-breast cancer activity of artemisinin and transferrin-loaded magnetic nanoliposome against breast transplanted tumors in BALB/c mice model. Materials and Methods: Artemisinin and transferrin-loaded magnetic nanoliposomes were prepared and characterized for some physiochemical properties. Pieces of tumor tissue from the breast cancer-bearing BALB/c mice were transplanted subcutaneously to the syngeneic female BALB/c mice. In the presence of the external magnet that placed at the breast tumor site, the tissue distribution and tumor-suppressing effects of prepared nanoliposomes on tumor growth was evaluated. Results: The prepared nanoliposomes have fine spherical shape, rough surface, nano-sized diameter and magnetic properties. At 2 h after treatment, the intravenous administration of artemisinin and transferrin-loaded magnetic nanoliposomes followed using the magnetic field approximately produced 10- and 5.5-fold higher levels of artemisinin and transferrin in the tumors, respectively, compared with free artemisinin and transferrin. Moreover, in the presence of an external magnetic field, the prepared nanoliposomes could significantly induce apoptosis in the mice breast cancer cells as well as could reduce tumor volume in tumorized mice at 15 days after treatment. Conclusion: The data suggested that the artemisinin and transferrin-loaded magnetic nanoliposomes would be a good choice for the breast tumor-targeted therapy, due to its high targeting efficiency.
  3 2,017 15
Pharmacological and phytochemical evaluation of Ocimum sanctum root extracts for its antiinflammatory, analgesic and antipyretic activities
Anant Kumar, Karishma Agarwal, Anil Kumar Maurya, Karuna Shanker, Umme Bushra, Sudeep Tandon, Dnyaneshwar U Bawankule
May 2015, 11(42):217-224
DOI:10.4103/0973-1296.157743  PMID:26109769
Background: Long-term use of nonsteroidal anti-inflammatory drugs (NSAIDs) increases risk of having a range of gastrointestinal problems. Therefore, new anti-inflammatory, analgesic, antipyretic drugs having lesser side effects are being searched all over the world as alternatives to NSAIDs. Aims: To evaluate the anti-inflammatory, analgesic and antipyretic profile of Ocimum sanctum root extracts. Materials and Methods: Anti-inflammatory profile of hexane (STH), chloroform (STC), ethyl acetate (STE), butanol (STB) and water (STW) extracts of OS was carried out by using carrageenan induced paw edema. STE a most active extract was further validated in dose dependent manner for anti-inflammatory, analgesic and antipyretic activity as well as oral toxicity profile in small laboratory animals. Identification of bioactives flux and chemical signature of most active fraction STE was developed by using the high-performance liquid chromatography fingerprinting. Results: An ethyl acetate fraction (STE) exhibit most potent anti-inflammatory activity followed by STB, STW, STC and STH. Dose response study of STE showed anti-inflammatory, analgesic and anti-pyretic potential in dose-dependent manner without any toxic effect at dose 2000 mg/kg. Chemical fingerprint revealed the presence of flavanoids. Conclusions: The present research revealed that STE possess anti-inflammatory, analgesic and anti-pyretic properties. However, future research is advocated to evaluate the pharmacological properties of isolated bioactive compounds.
  3 2,363 16
Cytotoxic activity of Macrosolen parasiticus (L.) Danser on the growth of breast cancer cell line (MCF-7)
Vijay Kumar Sodde, Richard Lobo, Nimmy Kumar, Rajalekshmi Maheshwari, CS Shreedhara
May 2015, 11(42):156-160
DOI:10.4103/0973-1296.157719  PMID:26109761
Background: Macrosolen parasiticus (L.) Danser belonging to Loranthaceaea (mistletoe family) is a parasitic plant that grows on different host plants such as mango, jack fruit, peepal, neem tree, etc., This study was aimed to investigate the anti-cancer activity of methanolic and aqueous extract of stem of M. parasiticus. Objectives: To investigate the in vitro cytotoxic potential of the methanolic and aqueous extracts from stems of M. parasiticus against MCF-7 breast cancer cells by brine shrimp lethality (BSL) bioassay, MTT assay and sulforhodamine B (SRB) assay. Materials and Methods: The extracts were tested in human breast cancer cell lines in vitro for percentage cytotoxicity, apoptosis by acridine orange/ethidium bromide staining, LD 50 and IC 50 values after treatment with M. parasiticus extracts. Results: In BSL bioassay, aqueous extract showed more significant (P < 0.01) cytotoxicity with LD 50 82.79 ± 2.67 μg/mL as compared to methanolic extract with LD 50 125 ± 3.04 μg/mL. The methanolic extract of M. parasiticus showed IC 50 97.33 ± 3.75 μg/mL (MTT) (P < 0.05) and 94.58 ± 3.84 μg/mL (SRB) (P < 0.01) assays against MCF-7. The aqueous extract of M. parasiticus demonstrated higher activity with IC 50 59.33 ± 3.3 μg/mL (MTT) (P < 0.01) and 51.9 ± 1.87 μg/mL (SRB) (P < 0.01) assays, after 48 h of exposure and thus showed significant dose-dependent cytotoxic activity. Conclusion: The finding demonstrated that both extracts of M. parasiticus showed significant cytotoxic activity, however aqueous extract demonstrated higher activity against MCF-7 breast cancer cells.
  2 1,939 16
Hepatorenal protective effect of Antistax ® against chemically-induced toxicity
Atallah F Ahmed, Hanan M Al-Yousef, Jawaher H Al-Qahtani, Mansour S Al-Said, AbdelKader E Ashour, Mohammed Al-Sohaibani, Syed Rafatullah
May 2015, 11(42):173-181
DOI:10.4103/0973-1296.157726  PMID:26109764
Background: Antioxidant natural products and chemoprevention are considered nowadays as an effective approach against health various disorders and diseases induced by oxidative stress or free radicals. Objective: The aim of this study was to assess the hepato- and nephroprotective activity of a standardized red vine leaf aqueous extract AS195 (Antistax ® ). Methods: The protective activity of AS195 (100 mg/kg) was investigated on carbon tetrachloride (CCl 4 )-intoxicated rats in comparison with silymarin. The flavonoid/proanthocyanidin nature of AS195 was identified by phytochemical and nuclear magnetic resonance (NMR) analyses, while its total phenol/proanthocyanidin/flavonoid content and antioxidant activity were determined by Folin-Ciocalteau, vanillin-sulfuric acid, AlCl 3 , and 2,2-diphenyl-2-picrylhydrazyl radical scavenging assays, respectively. Results: Relative to the control CCl 4 -intoxicated group, pretreatment with AS195 could significantly suppressed the elevated serum levels of alanine aminotransferase, alkaline phosphatase, γ-glutamyl transferase, total cholesterol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, triglycerides, bilirubin, creatinine, uric acid, and calcium, whereas it significantly increased the diminished serum levels of high-density lipoprotein cholesterol, albumin and total protein. Moreover, AS195 significantly decreased malondialdehyde formation in the tissues of liver and kidney, whereas it significantly elevated and nonprotein sulfhydryl groups, compared with the intoxicated control. The improvement in biochemical parameters by AS195 was obviously observed and further confirmed by restoration of normal histological features in the two organs. Conclusions: The results of the present study revealed the capacity of AS195 to enhance the recovery from xenobiotic-induced hepatorenal toxicity initiated by free radicals.
  2 1,835 16
Cytotoxic and apoptotic effects of Ebenus boissieri Barbey on human lung cancer cell line A549
Esra Arslan Aydemir, Ece Simsek, Nilüfer Imir, Ramazan Süleyman Göktürk, Erdem Yesilada, Kayahan Fiskin
May 2015, 11(42):37-45
DOI:10.4103/0973-1296.157679  PMID:26109772
Background: Fabaceae family members are known to possess preventive and therapeutic potentials against various types of cancers. Objective: The aim of this study was to investigate the cytotoxic and apoptotic effects of hydroalcoholic extracts from the aerial parts and roots of an endemic Ebenus species; Ebenus boissieri Barbey in human lung cancer cell line. Materials and Methods: After treatment with hydroalcoholic extracts cytotoxic activities of both extracts were measured by 3-(4, 5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay, whereas caspase-3 activity, tumor necrosis factor-alpha (TNF-α) and interferon gamma (IFN-γ) release were measured by enzyme linked immunosorbent assay. Results: According to in vitro assay results, the increase in all caspases activity suggested that extracts induce cells to undergo apoptosis. Especially, induction in caspase-3 activity was the most remarkable result of this study. Both aerial part and root extracts induced apoptosis by increasing caspase-3 activity, TNF-α and IFN-γ release. When compared to their relative controls, the concentrations of both TNF-α and IFN-γ in extract-treated groups were significantly and dose dependently exalted. Conclusion: Taken together, our results indicate that the hydroalcoholic extracts of E. boissieri can be considered as a source of new anti-apoptotic and therefore anti-carcinogenic agent.
  2 2,183 15
Anti-lung cancer potential of pure esteric-glycoside condurangogenin A against nonsmall-cell lung cancer cells in vitro via p21/p53 mediated cell cycle modulation and DNA damage-induced apoptosis
Sourav Sikdar, Avinaba Mukherjee, Anisur Rahman Khuda-Bukhsh
May 2015, 11(42):73-85
DOI:10.4103/0973-1296.157698  PMID:26109778
Background: Marsdenia condurango (condurango) is a tropical woody vine native to South America. Our earlier study was limited to evaluation of anti-cancer potentials of crude condurango extract and its glycoside-rich components in vitro on lung cancer. Objective: This study aims at evaluating the effect of the single isolated active ingredient condurangogenin A (ConA; C 32 H 42 O 7 ) on A549, H522 and H460-nonsmall-cell lung cancer cells. Materials and Methods: ConA was isolated by column chromatography and analyzed by mass spectroscopy, Fourier transform infrared spectroscopy and proton-nuclear magnetic resonance. diphenyltetrazolium bromide assays were conducted on three cell-types using 6%-alcohol as control. Critical studies on cellular morphology, cell-cycle regulation, reactive oxygen species, mitochondrial membrane potential, and DNA-damage were made, and expressions of related signaling markers studied. Results: As IC 50 doses of ConA proved to be too high and toxic to both A549 and H522 cells, all experimental studies were carried out on H460 cells with the IC 50 dose (32 μg/ml − 24 h). Cellular morphology revealed typical apoptotic features after ConA treatment. At early treatment hours (2 h-12 h), maximum cells were arrested at G0/G1 phase that could be correlated with reduced level of cyclin D1-CDK with p21 up-regulation. At 18 h − 24 h, sub G0/G1 cell population was increased gradually, as revealed from cytochrome-c release and caspase-3 activation, further confirming the apoptosis-inducing ability of ConA at later phases. Gradual increase of TUNEL-positive cells with significant modulation of mitochondria-dependent apoptotic markers at longer time-points would establish apoptosis-induction property of ConA, indicating its potential as a strong candidate for anti-cancer drug formulation. Conclusion: Further studies are warranted against other types of cancer cells and animal models before its possible human use.
  2 2,114 15
Toxicological and biochemical studies on Schinus terebinthifolius concerning its curative and hepatoprotective effects against carbon tetrachloride-induced liver injury
Rania H Abdou, Sherif Y Saleh, Waleed F Khalil
May 2015, 11(42):93-101
DOI:10.4103/0973-1296.157705  PMID:26109780
Background: Recently, many efforts have been made to discover new products of natural origin which can limit the xenobiotic-induced hepatic injury. Carbon tetrachloride (CCl 4 ) is a highly toxic chemical that is widely used to study hepatotoxicity in animal models. Objective: The present study was conducted to investigate the curative and protective effects of Schinus terbenthifolius ethanolic extract against CCl 4 -induced acute hepatotoxicity in rats. Materials and Methods: S. terbenthifolius extract was orally administered in a dose of 350 mg dried extract/kg b.wt. before and after intoxication with CCl 4 for curative and protective experiments, respectively. A group of hepatotoxicity indicative enzymes, oxidant-antioxidant capacity, DNA oxidation, and apoptosis markers were measured. Results: CCl 4 increased liver enzyme leakage, oxidative stress, hepatic apoptosis, DNA oxidation, and inflammatory markers. Administration of S. terebinthifolius, either before or after CCl 4 intoxication, significantly decreased elevated serum liver enzymes and reinstated the antioxidant capacity. Interestingly, S. terebinthifolius extract inhibited hepatocyte apoptosis as revealed by approximately 20 times down-regulation in caspase-3 expression when compared to CCl 4 untreated group. On the other hand, there was neither protective nor curative effect of S. terebinthifolius against DNA damage caused by CCl 4 . Conclusion: The present study suggests that S. terebinthifolius extract could be a substantially promising hepatoprotective agent against CCl 4 toxic effects and may be against other hepatotoxic chemical or drugs.
  2 1,689 15
Preconditioning with glycyrrhizic, ferulic, paeoniflorin, cinnamic prevents rat hearts from ischemia/reperfusion injury via endothelial nitric oxide pathway
Guo-Qiang Qian, Jingjing Ding, Xiaozhao Zhang, Xiaofeng Yin, Yuqin Gao, Guo-Ping Zhao
April-June 2015, 11(42):292-296
DOI:10.4103/0973-1296.153081  PMID:25829767
Objective: The objective was to investigate the endothelial nitric oxide synthase (eNOS/NO) pathway is involved or not in the protective effects of glycyrrhizic, ferulic, paeoniflorin, cinnamic (GFPC) in myocardial ischemia-reperfusion injury Sprague-Dawley rats. Materials and Methods: Ischemia-reperfusion (I/R) model was made by ligating the left anterior descending branch of the coronary artery for 30 min and releasing for 120 min, then the left ventricular apical was fixed and sliced, morphological changes of myocardial microvascular endothelial cell (MMVEC) was observed by electron microscopy, apoptosis index of MMVEC was observed by means of TUNEL, serum NO was tested by methods of nitrate reduction, lactate dehydrogenase (LDH), creatine kinase MB (CK-MB) was detected by automatic biochemical analyzer; Phosphorylated eNOS (PeNOS) and inducible NOS (iNOS) protein were measured by means of western blot. Results: In positive product control group, the serum levels of NO, LDH, CK-MB significantly increased (P < 0.05); MMVEC apoptosis was significantly decreased (P < 0.05); incidence of area at risk decreased significantly (P < 0.05); PeNOS protein increased (P < 0.05); iNOS protein decreased significantly (P < 0.05). Conclusion: Ischemic preconditioning of GFPC from GFPC plays a protective role in I/R heart through regulating the eNOS/NO signal pathway by increasing the PeNOS protein expression and decreasing the expression of iNOS protein.
  2 2,512 14
Anti-arthritic activity of Fu-Fang-Lu-Jiao-Shuang on collagen-induced arthritis in Balb/c mice and its underlying mechanisms
Yanyan Wang, Weiguang Sun, Laxia Chen, Xin Xu, Yunxia Wu, Jinwen Zhang, Yonghui Zhang
April-June 2015, 11(42):242-249
DOI:10.4103/0973-1296.153065  PMID:25829761
Background: Rheumatoid arthritis (RA) is a common, autoimmune disorder characterized by progressive multiple joint destruction, deformity, disability and premature death in most patients. Fu-Fang-Lu-Jiao-Shuang (FFLJS) is an effective traditional Chinese medicine, which has long been used clinically to treat RA patients. Objective: The objective of this study is aimed to evaluate the anti-rheumatic effects of FFLJS on collagen induced arthritis (CIA) model, as well as the underlying mechanisms, which have not previously been explored. Materials and Methods: CIA was induced by immunization with type II collagen (CII) in male Balb/c mice. The mice in the onset of arthritis were treated daily with FFLJS (125 or 500 mg/kg) or 1% carboxymethyl cellulose-Na for 28 days. Paw thickness and arthritic score were evaluated to confirm the anti-arthritic effect of FFLJS on CIA in mice. Levels of anti-CII antibody, proinflammatory cytokines interleukin-1 (IL-1) β, IL-17, and tumor necrosis factor-α (TNF-α) as well as prostaglandin E-2 (PGE-2) in serum and histological changes in the ankle joint were also analyzed. In addition, expressions of matrix metalloproteinases-1 (MMP-1), MMP-3 and tissue inhibitors of matrix metalloproteases-1 (TIMP-1) in synovial tissue were also detected to further study the molecular mechanism of the anti-arthritic effects of FFLJS. Results: During therapeutic treatment, FFLJS significantly reduced paw thickness and arthritic score in CIA mice, decreased the amounts of TNF-α, IL-1 β, IL-17, PGE-2 and anti-CII antibody in serum. In addition, FFLJS treatment could prevent the bone destruction by reducing the expression of MMP-1 and MMP-3, increasing the expression of TIMP-1 in synovial tissue of CIA mice. Conclusion: These findings offer the convincing evidence for the first time that the anti-rheumatic effects of FFLJS might be related to down-regulation of TNF-α, IL-1 β, IL-17 and PGE-2 levels for acute arthritis, and regulation of MMP-1, MMP-3 and TIMP-1 protein expression for chronic arthritis.
  2 3,662 15
Simultaneous quantification of six alkaloid components from commercial stemonae radix by solid phase extraction-high-performance liquid chromatography coupled with evaporative light scattering detector
Rong-Rong Zhang, Dan-Yi Lu, Zhen-Ya Yang, Wen Zhao, Paul Pui-Hay But, Pang-Chui Shaw, Ren-Wang Jiang, Zhi-Guo Ma
April-June 2015, 11(42):360-367
DOI:10.4103/0973-1296.153090  PMID:25829776
Background: Stemonae radix has been applied in traditional Chinese medicine for centuries. Alkaloids are the main active ingredient in stemonae radix, so their composition and concentration levels are directly linked to clinic effects. Objective: The objective was to develop an analytical method with multiple markers for quality survey of commercial stemonae radix. Materials and Methods: A method for simultaneous determination of six compounds in commercial stemonae radix was performed using solid-phase extraction and high-performance liquid chromatography coupled with evaporative light scattering detector. The separation was carried out on an Agilent TC-C18 column with 0.1% acetonitrile solution of triethylamine aqueous solution and acetonitrile as the mobile phase under gradient elution within 70 min. The hierarchical clustering analysis (HCA) was successfully used to classify the samples in accordance with their chemical constituents. Results: Linearity (R 2 > 0.9990), intra- and inter-day precision (relative standard deviations <4%), limit of detection (0.011-0.086 μg/mL), limit of quantification (0.033-0.259 μg/mL) of the six alkaloids were determined, and the recoveries were between 96.6% and 103.7%. The method was successfully applied to analysis 36 batches of commercial stemonae radix. All the samples could be classified into five clusters by HCA. Conclusion: This article provides an accurate and simple analytical method for quality survey of commercial stemonae radix. Because of the significant chemical variations, careful selection of Stemona sources with obvious antitussive value but devoid of croomine followed by good agricultural practice and good manufacturing practice process is suggested.
  2 2,403 16
Development and extraction optimization of baicalein and pinostrobin from Scutellaria violacea through response surface methodology
Shankar Subramaniam, Ravikumar Raju, Anbumathi Palanisamy, Aravind Sivasubramanian
May 2015, 11(42):127-138
DOI:10.4103/0973-1296.157714  PMID:26109758
Objective: To develop a process that involves optimization of the amount of baicalein and pinostrobin from the hydro-methanolic extract of the leaves of Scutellaria violacea by response surface methodology (RSM). Materials and Methods: The combinatorial influence of various extraction parameters on the extraction yield was investigated by adopting Box-Behnken experimental design. Preliminary experiments carried out based on the traditional one variable at a time optimization revealed four such operational parameters to play a crucial role by influencing the yield. These four process parameters at three levels were considered to obtain the Box-Behnken experimental design. Results: RSM based model fitted to the resulting experimental data suggested that 52.3% methanol/water, 12.46:1 solvent-solid ratio, 285 rpm agitation and 6.07 h of extraction time are the optimal conditions which yielded a maximized amount of baicalein and pinostrobin of 2.9 and 4.05 mg/g DM. Analysis of variance revealed a high correlation coefficient (R2 = 0.999 for baicalein and 0.994 for pinostrobin), signifying a good fit between the regression model (second order) and the experimental observations. Conclusion: The present study signifies that both the metabolites have been extracted from S. violacea for the first time. Further, this study developed an optimized extraction procedure to obtain maximum yield of the metabolites, which is unique and better than conventional extraction methodology. The operational parameters under optimized conditions accounts for the lowest cost in extraction process thus, providing an efficient, rapid and cost-effective method for isolation and scale up of these commercially vital flavonoids.
  2 1,734 15
Methanol extract of wheatgrass induces G1 cell cycle arrest in a p53-dependent manner and down regulates the expression of cyclin D1 in human laryngeal cancer cells-an in vitro and in silico approach
Garima Shakya, Sangeetha Balasubramanian, Rukkumani Rajagopalan
May 2015, 11(42):139-147
DOI:10.4103/0973-1296.157715  PMID:26109759
Background: Deregulation of cell cycle has been implicated in the malignancy of cancer. Since many years investigation on the traditional herbs has been the focus to develop novel and effective drug for cancer remedies. Wheatgrass is a medicinal plant, used in folk medicine to cure various diseases. The present study was undertaken to gain insights into antiproliferative effect of methanol extract of wheatgrass. Materials and Methods: Cell viability was assessed via 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Lactate Dehydrogenase assays. Cell cycle was analyzed by flow cytometry. Western blot was performed to determine the p53 and cyclin D1 levels. In silico docking interaction of the 14 active components (identified by high-performance liquid chromatography/gas chromatography-mass spectroscopy) of the methanol extract was tested with cyclin D1 (Protein Data Bank ID: 2W96) and compared with the reference cyclin D1/Cdk4 inhibitor. Results: Methanol extract of wheatgrass effectively reduced the cell viability. The cell cycle analysis showed that the extract treatment caused G 1 arrest. The level of cyclin D1 was decreased, whereas p53 level was increased. Molecular docking studies revealed interaction of seven active compounds of the extract with the vital residues (Lys112/Glu141) of cyclin D1. Conclusion: These findings indicate that the methanol extract of wheatgrass inhibits human laryngeal cancer cell proliferation via cell cycle G 1 arrest and p53 induction. The seven active compounds of the extract were also found to be directly involved in the inhibition of cyclin D1/Cdk4 binding, thus inhibiting the cell proliferation.
  2 1,928 14
Cytotoxic glucosphingolipid from Celtis Africana
Shagufta Perveen, Areej Mohammad Al-Taweel, Ghada Ahmed Fawzy, Azza Muhammed El-Shafae, Afsar Khan, Peter Proksch
May 2015, 11(42):1-5
DOI:10.4103/0973-1296.157662  PMID:26109753
Background: Literature survey proved the use of the powdered sun-dried bark and roots of Celtis africana for the treatment of cancer in South Africa. Objective: The aim of this study was to do further isolation work on the ethyl acetate fraction and to investigate the cytotoxic activities of the various fractions and isolated compound. Materials and Methods: Cytotoxicity of petroleum ether, chloroform, ethyl acetate, n-butanol fractions and compound 1 were tested on mouse lymphoma cell line L5178Y using the microculture tetrazolium assay. Results: One new glucosphingolipid 1 was isolated from the aerial parts of C. africana. The structure of the new compound was determined by extensive analysis by one-dimensional and two-dimensional nuclear magnetic resonance spectroscopy and mass spectrometry. The ethyl acetate fraction and compound 1 showed strong cytotoxic activity with an EC 50 value of 8.3 μg/mL and 7.8 μg/mL, respectively, compared with Kahalalide F positive control (6.3 μg/mL). Conclusion: This is the first report of the occurrence of a cytotoxic glucosphingolipid in family Ulmaceae.
  2 2,046 16
Yin-Chen-Hao-Tang alleviates biliary obstructive cirrhosis in rats by inhibiting biliary epithelial cell proliferation and activation
Bing Wang, Ming-Yu Sun, Ai-Hua Long, Hong-Yan Cao, Shuang Ren, Yan-Qin Bian, Xiong Lu, Hong-Tu Gu, Cheng-Hai Liu, Ping Liu
April-June 2015, 11(42):417-425
DOI:10.4103/0973-1296.153098  PMID:25829784
Background: Yin-Chen-Hao-Tang (YCHT) consists of three aqueous extracts from Artemisia capillaris, Gardenia sp., and prepared Rheum rhabarbarum (rhubarb) (3:2:1). YCHT is characterized by its anti-inflammatory properties in liver regulation and relief of jaundice. We aimed to study the effects and mechanisms of action of YCHT on biliary obstructive cirrhosis. Materials and Methods: Secondary biliary fibrosis was induced in rats by bile duct ligation (BDL) and scission. One week after BDL, rats were randomly divided into a saline-treated BDL or YCHT-treated BDL group for 4 weeks. Liver function and hepatic hydroxyproline (Hyp) content were assessed. Types I and IV collagen (Col-IV), laminin, fibronectin, alpha smooth muscle actin (α-SMA), and proliferating cell nuclear antigen protein and messenger ribonucleic acid (mRNA) expression were assessed with immunohistochemistry and real-time polymerase chain reaction. Results: In the YCHT-treated BDL group, serum total bilirubin, total bile acids, aspartate aminotransferase, alanine aminotransferase, and g-glutamyl transferase were lower than those in the sham-operated BDL group. The proliferation of bile ducts in hepatic tissues and the Hyp content and Col deposition were also significantly lower than those in control rats. In addition, α-SMA and Col-IV staining was less obvious, and mRNA expression of Procol-α1 (IV), platelet derived growth factor subunit B (PDGF)-B, connective tissue growth factor, and transforming growth factor-beta in proliferative biliary epithelial cells (BECs) in the YCHT-treated BDL group was significantly lower than those in controls. Conclusions: YCHT effectively reduces the formation of biliary obstructive cirrhosis mainly via inhibition of BEC proliferation by down-regulation of PDGF-B mRNA expression, inhibition of BEC profibrogenic paracrines, and the epithelial-mesenchymal transition pathological process.
  2 2,812 16
Effects of Pogostemon cablin Blanco extract on hypoxia induced rabbit cardiomyocyte injury
Chi-Yeon Lim, Bu-Yeo Kim, Se-Hyun Lim, Su-In Cho
April-June 2015, 11(42):311-319
DOI:10.4103/0973-1296.153084  PMID:25829770
Background: Pogostemonis Herba, the dried aerial part of Pogostemon cablin Blanco, is a well-known materia medica in Asia that is widely used for syndrome of gastrointestinal dysfunctions. Objective: This study was undertaken to examine whether Pogostemon cablin extract (PCe) might have any beneficial effect on hypoxia induced rabbit cardiomyocyte injury. Materials and Methods: Isolated cardiomyocytes were divided into three groups and the changes of cell viability in cardiomyocytes of hypoxic and hypoxia/reoxygenation group were determined. The effect of PCe on reactive oxygen species (ROS) generation, intracellular formation of ROS was also measured by monitoring the 2',7'- dichlorofluorescein fluorescence. Results: PCe effectively protected the cells against both the hypoxia and reoxygenation induced injury, and the protective effect of PCe is not mediated by interaction with adenosine triphosphate-sensitive K + channels. In the presence of PCe, production of ROS under chemical hypoxia was remarkably reduced which suggests that PCe might exert its effect as a ROS scavenger. Conclusion: The present study provides clear evidence for the beneficial effect of PCe on cardiomyocyte injury during hypoxia or reoxygenation following prolonged hypoxia.
  2 2,508 14
Antiradical and antidiabetic properties of standardized extract of Sunderban mangrove Rhizophora mucronata
Tapas Kumar Sur, Alok Kumar Hazra, Dipankar Bhattacharyya, Avijit Hazra
April-June 2015, 11(42):389-394
DOI:10.4103/0973-1296.153094  PMID:25829780
Background: Mangroves have the ability to grow where no other vascular plants survive. Rhizophora mucronata is a true mangrove and traditionally used to treat diabetes and its allied complications. Objectives: In the present study, we standardized the 80% methanolic standardized extract of R. mucronata leaves (RH) and found out its antiradical and antidiabetic activities. Materials and Methods: The methanolic extract of R. mucronata leaves (RH) was standardized and quantified for phenolics, flavonoinds, gallic acid, quercetin, and coumarin. The reducing abilities and antiradical activities of RH were performed in vitro methods like, 1,1-diphenyl-2-picrylhydrazyl, nitric oxides, superoxides, hydroxyl, and ABTS (2,2'-azino-bis-3-ethyl benzthiazoline-6-sulphonic acid). Thereafter, RH was evaluated for it antidiabetic potentialities on streptozotocin (STZ)-induced type-2 diabetes. STZ (90 mg/kg, intraperitoneal) was administered to 2 days old pups to induce diabetes. RH was fed at doses of 50 and 100 mg/kg and glibenclamide (positive control) at 5 mg/kg, when the rats were 6 weeks old and continued for 10 weeks. Fasting glucose was monitored before and after the treatment. Further, lipid peroxides and reduced glutathione level were estimated on rat liver. Results: The results obtained from this study revealed RH possesses flavonoinds and also gallic acid, quercetin, and coumarin. Further, it has antiradical activities. It has also reduced blood glucose level in type-2 diabetic rats and reduced the formation of lipid peroxidation in liver. RH enhanced the level of glutathione in liver tissue. Conclusion: RH exhibits source of natural antioxidants and great potentialities as an antidiabetic agent by improving the hyperglycemia through its antiradical action.
  2 2,999 14
Effect of co-administration of Angelicae gigantis radix and Lithospermi radix on rat hepatic injury induced by carbon tetrachloride
Chi-Yeon Lim, Bu-Yeo Kim, Se-Hyun Lim, Su-In Cho
April-June 2015, 11(42):395-403
DOI:10.4103/0973-1296.153095  PMID:25829781
Background: Co-administration of Angelicae gigantis radix (AGR) and Lithospermi radix (LR) has been commonly applied to patients to treat cardiac and hepatic disorders. Individual bioactivities of these herbal medicines have been widely investigated, but the hepatoprotective effects of co-treatment of AGR and LR have yet to be clarified. Objective: The present study investigated the protective effects of extracts of AGR and LR on carbon tetrachloride (CCl 4 ) induced hepatic injury. Materials and Methods: In this study, we measured the hepatoprotective activity of individual and co-treatment of the two herbal medicines on hepatic injury induced by CCl 4 by measuring different biochemical parameters such as serum aspartate aminotransaminase (AST) and serum alanine aminotransaminase (ALT). Microarray technology also used to compare ontological difference with individual and co-treatment of these two. Results: Combined treatment with AGR and LR (AGR + LR) decreased AST and ALT level in serum which demonstrate hepatoprotective effect of the therapy. When the effect of AGR and LR according to treatment conditions was measured, co-treatment showed the most prominent effect on hepatic injury by CCl 4 rather than individual treatment condition. We further defined gene set that could be the molecular target of herbal effect on hepatic injury by CCl 4 using bioinformatical analysis of interaction network. Highly recovered genes by treating AGR + LR play significant roles in response to hepatic injury induced by CCl 4 . Conclusion: Combined treatment with AGR and LR showed synergistic protective effects on the CCl 4 -induced rat hepatic tissue injury.
  1 2,545 15
Quantitative analysis of the eight major compounds in the Samsoeum using a high-performance liquid chromatography coupled with diode array detection and electrospray ionization mass spectrometer
Jin Bae Weon, Hye Jin Yang, Bohyoung Lee, Jin Yeul Ma, Choong Je Ma
April-June 2015, 11(42):320-326
DOI:10.4103/0973-1296.153085  PMID:25829771
Background: Samsoeum was traditionally used for treatment of a respiratory disease. Objective: The simultaneous determination of eight major compounds, ginsenoside Rg3, caffeic acid, puerarin, costunolide, hesperidin, naringin, glycyrrhizin, and 6-gingerol in the Samsoeum using a high-performance liquid chromatography (HPLC) coupled with diode array detection (DAD) and an electrospray ionization mass spectrometer was developed for an accurate and reliable quality assessment. Materials and Methods: Eight compounds were qualitative identified based on their mass spectra and by comparing with standard compounds and quantitative analyzed by HPLC-DAD. Separation of eight compounds was carried out on a LUNA C 18 column (S-5 μm, 4.6 mm i.d. ×250 mm) with gradient elution composed of acetonitrile and 0.1% trifluoroacetic acid. Results: The data showed good linearity (R2 > 0.9996). The limits of detection and the limits of quantification were <0.53 μg and 1.62 μg, respectively. Inter- and Intra-day precisions (expressed as relative standard deviation values) were within 1.94% and 1.91%, respectively. The recovery of the method was in the range of 94.24-107.90%. Conclusion: The established method is effective and could be applied to quality control of Samsoeum.
  1 2,564 15
Attenuation of reactive oxygen/nitrogen species with suppression of inducible nitric oxide synthase expression in RAW 264.7 macrophages by bark extract of Buchanania lanzan
Shailasree Sekhar, KK Sampath-Kumara, SR Niranjana, HS Prakash
April-June 2015, 11(42):283-291
DOI:10.4103/0973-1296.153080  PMID:25829766
Background: Oxidative stress is one of the most critical factors implicated in disease conditions. Buchanania lanzan Spr. (Anacardiaceae) bark powder preparation has been reported for treating an inflammatory condition in the Ayurvedic Pharmacopoeia of India. Objective: In the present study, we investigate the effect of the bark methanol extract (BLM) on reactive oxygen/nitrogen species (ROS/RNS), the expression of protein and mRNA of inducible nitric oxide synthase (iNOS) in RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS) and sodium nitroprusside (SNP) to provide scientific validation of the above said medicinal property. Materials and Methods: The capacity to quench ROS and RNS was evaluated by 5-(and-6) chloromethyl-20,70-dichlorodihydrofluorescein diacetate acetyl ester fluorescence and nitrite estimations in LPS/SNP-stimulated macrophages respectively. The protein and transcript expression of iNOS was evaluated through Western Blot and reverse transcription-polymerase chain reaction (RT-PCR) analysis respectively. Results: Macrophages pretreated with BLM (>100 μg/mL) for 24 h, stimulated with LPS for the last 18 h of experimental duration recorded a significantly (P < 0.05) reduced levels of ROS (3.45-fold) against LPS-stimulated conditions (5.7-fold). SNP-stimulation resulted in increased NO accumulation (17-fold) which was neutralized by BLM at >100 μg/ml (1.6-fold) credited to a reduced protein and mRNA expression of iNOS as recorded by Western blots and RT-PCR results respectively. The reversed-phase liquid chromatography-diode array detection analysis identified the presence of 4-hydroxybenzoic acid, quercetin and p-coumaric acid (Rt values 5.444, 5.569 and 9.580 respectively). Conclusions: The potential of BLM inhibiting ROS/RNS production validates the medical use of bark, could find beneficial application under conditions of immune stimulation and/or bacterial infection.
  1 2,930 15
Inhibitory potential of some Romanian medicinal plants against enzymes linked to neurodegenerative diseases and their antioxidant activity
Gabriela Paun, Elena Neagu, Camelia Albu, Gabriel Lucian Radu
May 2015, 11(42):110-116
DOI:10.4103/0973-1296.157709  PMID:26109755
Context: Eryngium planum, Geum urbanum and Cnicus benedictus plants are an endemic botanical from the Romanian used in folk medicine. Objective: The extracts from three Romanian medicinal plants were investigated for their possible neuroprotective potential. Materials and Methods: Within this study, in vitro neuroprotective activity of the extracts of E. planum, G. urbanum, and C. benedictus plants were investigated via inhibition of acetylcholinesterase (AChE) and tyrosinase (TYR). Total content of phenolics, flavonoids, and proanthocyanidins, high-performance liquid chromatography profile of the main phenolic compounds and antioxidant activity were also determined. Results: Among the tested extracts, the best inhibition of AChE (88.76 ± 5.2%) and TYR (88.5 ± 5.2%) was caused by C. benedictus ethanol (EtOH) extract. The G. urbanum extracts exerted remarkable scavenging effect against 2,2-diphenyl-1-picrylhydrazyl (IC 50 , 7.8 ± 0.5 μg/mL aqueous extract, and IC 50 , 1.3 ± 0.1 μg/mL EtOH extract, respectively) and reducing power, whereas the EtOH extract of C. benedictus showed high scavenging activity (IC 50 , 0.609 ± 0.04 mg/mL), also. Conclusion: According to our knowledge, this is the first study that demonstrates in vitro neuroprotective effects of E. planum, G. urbanum and C. benedictus.
  1 1,656 16
The dose dependent in vitro responses of MCF-7 and MDA-MB-231 cell lines to extracts of Vatica diospyroides symington type SS fruit include effects on mode of cell death
Theera Srisawat, Yaowapa Sukpondma, Potchanapond Graidist, Siriphon Chimplee, Kanyanatt Kanokwiroon
May 2015, 11(42):148-155
DOI:10.4103/0973-1296.157718  PMID:26109760
Background: Vatica diospyroides type LS is a known source of valuable compounds for cancer treatment, however, in contrast little is known about therapeutic efficacy of type SS. Objective: This study focused on in vitro cytotoxicity of these fruit extracts, and the cell death mode they induce in breast cancer cells. Materials and Methods: Acetone extracts of fruit were tested for cytotoxicity against MCF-7 and MDA-MB-231 cell lines. The apoptosis and necrosis of these cells were quantified by fluorescence activated cell sorter (FACS) and western blot analyses. Results: After 72 h of treatment, the 50% growth inhibition concentrations (IC 50 ) levels were 16.21 ± 0.13 µg/mL against MCF-7 and 30.0 ± 4.30 µg/mL against MDA-MB-231, indicating high and moderate cytotoxicity, respectively. From the FACS results, we estimate that the cotyledon extract at half IC 50 produced 11.7% dead MCF-7 cells via apoptosis, whereas another concentrations both apoptosis and necrosis modes co-existed in a dose-dependent manner. In MDA-MB-231 cell line, only the apoptosis was induced by the pericarp extract in a dose-dependent manner. With the extracts at half IC 50 concentration, in both cells, the expression of p21 decreased while that of Bax increased within 12-48 h of dosing, confirming apoptosis induced by time-dependent responses. Apoptosis dependent on p53 was found in MCF-7, whereas the mutant p53 of MDA-MB-231 cells was expressed. Conclusion: The results indicate that fruit extracts of V. diospyroides have cytotoxic effects against MCF-7 and MDA-MB-231 cells via apoptosis pathway in a dose-dependent manner. This suggests that the extracts could provide active ingredients for the development, targeting breast cancer therapy.
  1 1,639 14
Evaluation of extraction method on the chemical composition in Apeiba tibourbou Aubl's extracts
Frederico Severino Martins, Edemilson Cardoso da Conceição
April-June 2015, 11(42):368-373
DOI:10.4103/0973-1296.153091  PMID:25829777
Background: The extraction method of bioactive compounds is an important step in the manufacturing of herbal medicines, because secondary metabolites with therapeutic potential are usually found in small quantities in plant materials. Objective: Due the potential of Apeiba tibourbou Aubl, this study aimed to evaluate the impact of the extraction method on the quality of herbal extract and optimize the extraction of fatty acid, rosmarinic (Ra) and caffeic (Ca) acid from A. tibourbou. Materials and Methods: Determinations of residual moisture (Rm), proteins (Pt), lipids (Lp), total fiber (Tf), and carbohydrate (Cy) were performed in triplicate samples according assessment of antioxidant capacity. Extraction of fatty acids was carried out by two different methods: (i) By shoxlet and (ii) bligh and dyer. The optimized conditions were determined by surface response methodology (RSM), and the criterion of desirability was the maximum extraction of Ra and Ca. Results: The method of bligh and dyer was able to extraction more total Lp than the shoxlet. However, the extraction of fatty acid was different for the two methods. The optimized conditions to extract RA and Ca was calculated by RSM, 42°C, 30% (alcohol degree) and 24 min, this conditions maximize simultaneously the extraction of Ca (0, 04%) and Ry (1.89), Conclusion: It was observed that the extraction method alters the chemical composition of extract, and it is possible to extract Ca and Ra from A. tibourbou's leaves using ultrasound-assisted extraction.
  1 2,518 14
In vitro regeneration and Agrobacterium tumefaciens-mediated genetic transformation in asakura-sanshoo (Zanthoxylum piperitum (L.) DC. F. inerme Makino) an important medicinal plant
Xiaofang Zeng, Degang Zhao
April-June 2015, 11(42):374-380
DOI:10.4103/0973-1296.153092  PMID:25829778
Context: Asakura-sanshoo (Zanthoxylum piperitum [L.] DC. f. inerme Makino) is an important medicinal plant in East Asia. Transgenic technique could be applied to improve plant traits and analyze gene function. However, there is no report on regeneration and genetic transformation in Asakura-sanshoo. Aims: To establish a regeneration and Agrobacterium tumefaciens-mediated genetic transformation system in Asakura-sanshoo, which could be used for cultivar improvement and gene function analysis. Settings and Design: The various combinations of indole-3-butyric acid (IBA), 6-benzylaminopurine (BA) and naphthalene acetic acid (NAA) were explored for the optimal plant regeneration from petiole and stem of Asakura-sanshoo. The half-strength woody plant medium (WPM) with different concentrations of NAA and IBA was used to induce root. For genetic transformation, A. tumefaciens strain EHA-105 harboring the plasmid pBin-Ex-H-ipt which carries the isopentenyl transferase (ipt) gene, β-glucuronidase (GUS) gene and kanamycin resistance gene neomycin phosphotransferase II (NPTII) were used. The transformation efficiency was detected by the kanamycin resistant frequency. Materials and Methods: Petioles and stems were obtained from the in vitro cultured Asakura-sanshoo. The petiole and stem segments were precultured for 3 days, and then inflected using the bacterium at the concentration of OD 600 0.5-0.8 for 10 min, followed by 3 days co-cultivation. Selection of the transgenic plants was carried out after 7 days the regeneration using gradient kanamycin at 30 mg/L and 50 mg/L, respectively. Successful transformed plants were confirmed by GUS histochemical assays, polymerase chain reaction (PCR), reverse transcription-PCR (RT-PCR), and Southern blotting analysis. Results: The highest shoots regeneration was obtained on WPM supplement with 0.5 mg/L BA and 0.2 mg/L NAA. The optimal rooting medium was half strength macro-element WPM. The kanamycin resistant frequency of petiole and stem was 24.66% and 25.93%, respectively. Thirty-five shoots in thousands adventitious buds were confirmed through GUS histochemical assays, PCR, RT-PCR, and Southern blotting. The regeneration shoot per explants elevated 5.85 fold compared with the wild-type plants. Conclusions: Individual transgenic Asakura-sanshoo lines were obtained. In this paper, it first revealed the expression of ipt gene significantly promoted the adventitious buds induction in Asakura-sanshoo as the same action as in other plants.
  1 3,231 16
The influences of inorganic elements in soil on the development of famous - region Atractylodes lancea (Thunb.) DC
Weiwanqi Zhang, Zhen Ouyang, Ming Zhao, Yuan Wei, Huasheng Peng, Qiang Wang, Ling Guo
April-June 2015, 11(42):337-344
DOI:10.4103/0973-1296.153087  PMID:25829773
Background: Atractylodes lancea (Thunb.) DC., is an important medicinal plant in China. Recently, researches of A. Lancea were focused on chemical composition and genetics, only a few were concerned with soil factors. Objective: The aim was to discuss the relationship between geo-herbalism of A. Lancea (Thunb.) DC. and inorganic elements in soil. Materials and Methods: The contents of 15 kinds of inorganic elements in the rhizoma of A. Lancea (Thunb.) DC. and soils from various regions were determined with inductively coupled plasma-optical emission spectrometer and the data were analyzed with Statistical Package for the Social Sciences 20.0 software. Results: The contents of inorganic elements in rhizoma of A. Lancea and in soil with different geological background were different. The soils in the famous region contained high aluminum, iron, sodium and low sulfur content. The rhizoma of A. Lancea contained high aluminum, lithium, manganese and low iron, sulfur content. The famous-region crude drugs had a strong tendency to accumulate selenium, manganese. Ten characteristic elements of A. Lancea were K, Ca, S, Al, Li, Ti, Mn, Pb, Ni, SE. Conclusion: The contents of inorganic elements in rhizoma of A. Lancea showed a significant positive interrelationship with those in soil. It was identified that inorganic elements play an important role in forming authenticity A. Lancea (Thunb.) DC.
  1 2,529 15
Ethyl acetate extract from marine sponge Hyattella cribriformis exhibit potent anticancer activity by promoting tubulin polymerization as evidenced mitotic arrest and induction of apoptosis
Pazhanimuthu Annamalai, Malini Thayman, Sowmiya Rajan, Lakshmi Sundaram Raman, Sankar Ramasubbu, Pachiappan Perumal
April-June 2015, 11(42):345-355
DOI:10.4103/0973-1296.153088  PMID:25829774
Background: Marine sponges are important sources of bioactive compounds. Objective: This study investigated the anticancer properties of Hyattella cribriformis ethyl acetate (EA) fraction in various cancer and normal cell lines. Materials and Methods: anticancer assay was carried out in 15 cell lines to evaluate the anticancer potential of the EA fraction. Impact on cell cycle distribution was determined using flow cytometry. The fraction was investigated for interfering microtubules assembly in both in vitro and cellular assay. Further studies were conducted to determine the fraction induced cell death (apoptosis) using calcein/propidium iodide dual staining, activated caspase-3 and phosphorylation of Bcl-2 protein at Ser70. DNA fragmentation assay was performed to confirm the apoptosis. Results: EA fraction exhibited potent inhibition of cancer cell growth and resulted in 50% growth inhibition (GI 50 ) of 0.27 μg/mL in A673 cell line. Sarcoma (MG-63, Saos-2) and ovarian (SK-OV-3 and OVCAR-3) cancer cell lines also showed superior anticancer activity GI 50 of 1.0 μg/mL. Colon and breast cancer cell lines exhibited moderate GI compare other cancer cell lines and normal human lung fibroblast showed GI 50 of 15.6 μg/mL. EA fraction showed potent G2/M phase arrest in A673 cell line and induced apoptosis at 48 h exposure. EA fraction promoted microtubule polymerization in tubulin polymerization assay and increased level of polymerized tubulin in the HeLa cells. Fraction induced the activation of caspase-3 and phosphorylation of Bcl-2 anti-apoptotic protein. Fraction induced DNA fragmentation in HeLa cells as evidence of apoptosis. Conclusion: Marine sponge H. cribriformis EA fraction exhibited potent anticancer activity through tubulin polymerization and induction of apoptosis.
  1 3,156 14
Identification of longevity, fertility and growth-promoting properties of pomegranate in Caenorhabditis elegans
Hasan Kiliçgün, Nazli Arda, Evren Önay Uçar
April-June 2015, 11(42):356-359
DOI:10.4103/0973-1296.153089  PMID:25829775
Background: Pomegranate (Punica granatum L.) is commonly consumed as fresh fruit and fruit juice. It is also used in the production of jam, wine, food coloring agent, and flavor enhancer. Objective: The aim of this study was to identify the possible longevity, fertility and growth promoting properties of different ethanolic extract concentrations of pomegranate in Caenorhabditis elegans, which is increasingly popular and has proven to be a very useful experimental model organism for aging studies as well as for testing antioxidants and other compounds for effects on longevity. Materials and Methods: In this study, five experimental groups (20, 10, 5, 2.5 and 1.25 mg pomegranate extract/mL and one control group) were used to determine the most effective dose of pomegranate in terms of longevity, fertility and growth parameters. Results: It was seen that, pomegranate extracts up to the concentration of 5 mg/mL, had the potential to promote for the longevity, formation of new generations, fertility of new generations and growth properties of C. elegans although higher concentrations significantly reduced these parameters. Conclusion: these findings indicated that pomegranate could be used as a supplement to enhance longevity, fertility and growth rate for the other living organisms and human beings, but the dose should be carefully adjusted to avoid adverse effects.
  1 895 5
The enriched fraction of Elephantopus scaber Triggers apoptosis and inhibits multi-drug resistance transporters in human epithelial cancer cells
Asmy Appadath Beeran, Naseer Maliyakkal, Chamallamudi Mallikarjuna Rao, Nayanabhirama Udupa
April-June 2015, 11(42):257-268
DOI:10.4103/0973-1296.153077  PMID:25829763
Background: Medicinal plants have played an important role in the development of clinically useful anticancer agents. Elephantopus scaber (Asteraceae) (ES) is widely used in Indian traditional system of medicine for the treatment of various ailments including cancer. Objective: To investigate anticancer effects of ES in human epithelial cancer cells. Materials and Methods: Cytotoxicity of ethanolic extract of ES (ES-ET) and its fractions, such as ES Petroleum ether fraction (ES-PET), ES Dichloromethane fraction (ES DCM), n Butyl alcohol fraction (ES-BT), and ES-Rest (ES-R) were assessed in human epithelial cancer cell lines using sulforhodamine B (SRB) assay. Acridine orange/ethidium bromide assay and Hoechst 33342 assays were used to gauge induction of apoptosis. Cell cycle analysis and micronuclei assay were used to assess cell cycle specific pharmacological effects and drug induced genotoxicty. Further, the ability of ES to inhibit multi drug resistant (MDR) transporters (ABC-B1 and ABC-G2) was determined by Rhodamine (Rho) and Mitoxantrone (MXR) efflux assays. Results: The enriched fraction of ES (ES DCM) possessed dose-dependent potent cytotoxicity in human epithelial cancer cells. Further, treatment of cancer cells (HeLa, A549, MCF-7, and Caco-2) with ES DCM showed hall mark properties of apoptosis (membrane blebbing, nuclear condensation etc.). Similarly, ES DCM caused enhanced sub G0 content and micronuclei formation indicating the induction of apoptosis and drug induced genotoxicity in cancer cells, respectively. Interestingly, ES DCM inhibited MDR transporters (ABC B1 and ABC G2) in cancer cells. Conclusion: The enriched fraction of ES imparted cytotoxic effects, triggered apoptosis, induced genotoxicity, and inhibited MDR transporters in human epithelial cancer cells. Thus, ES appears to be potential anticancer agent.
  1 3,246 14
Protective effect of young green barley leaf (Hordeum vulgare L.) on restraint stress-induced decrease in hippocampal brain-derived neurotrophic factor in mice
Katsunori Yamaura, Riho Tanaka, Yuanyuan Bi, Hideki Fukata, Nobuo Oishi, Hiromi Sato, Chisato Mori, Koichi Ueno
May 2015, 11(42):86-92
DOI:10.4103/0973-1296.157702  PMID:26109779
Background: Many health experts support the hypothesis that stressful lifestyles are the leading cause of illness, like depression. Therefore, from the standpoint of preventive medicine, it is important to reduce stress. Young green barley leaves are a good natural source of vitamins and minerals, and their juice is widely consumed as a functional food for health reasons in Japan. This study investigated the protective effect of young green barley leaves for stress control. Materials and Methods: ICR outbred mice were exposed to 3-h sessions of restraint stress. Young green barley leaves (400 and 1,000 mg/kg) were administered orally 1 h before the sessions for 5 days. To analyze voluntary behavior, wheel-running activity was monitored during the dark period. Brain-derived neurotrophic factor (BDNF) messenger RNA (mRNA) expression in the whole hippocampus was measured by real-time quantitative polymerase chain reaction. Results: Restraint stress resulted in a significant decrease in voluntary wheel-running behavior, but this decrease was ameliorated by the administration of young green barley leaves. The leaves also enhanced the decreased levels of BDNF mRNA induced by restraint stress; in particular, a significant protective effect was shown in the exon IV variant as compared to vehicle control mice. Conclusion: The findings suggest that young green barley leaves have potent anti-stress properties, as evidenced by preventing decreases in the levels of voluntary wheel-running activity and hippocampal BDNF mRNA in response to restraint stress. Our findings support the possibility that supplementation with young green barley leaves might be beneficial for preventing stress-related psychiatric disorders like depression.
  1 1,653 15
Antimutagenic potential of harpagoside and Harpagophytum procumbens against 1-nitropyrene
Luigi Manon, Baghdikian Béatrice, Orsière Thierry, Pompili Jocelyne, Mabrouki Fathi, Ollivier Evelyne, Botta Alain
May 2015, 11(42):29-36
DOI:10.4103/0973-1296.157675  PMID:26109771
Background: 1-nitropyrene (1-NPy) is one of the most abundant nitro-polycyclic aromatic hydrocarbons particularly in diesel exhausts. It is a mutagenic and carcinogenic pollutant very widespread in the environment. So the discovery of antimutagenic agents is essential. Harpagophytum procumbens (HP) is traditionally used as anti-inflammatory and analgesic particularly against painful osteoarthritis. Harpagoside (HS), its major iridoid glycoside, is considered as the main active component. Objective: The aim of the present study was to evaluate the antimutagenic activity of HS and HP extracts against mutagenic activity of 1-NPy. Materials and Methods: The antimutagenic activity was investigated using the in vitro cytokinesis-block micronucleus assay in cultured human lymphocytes. Cells were exposed to HS or HP extracts before (pretreatment), during (co-treatment), and after (posttreatment) treatment with 1-NPy. Results: Results showed that HS significantly reduced the mutagenicity of 1-NPy in pretreatment and particularly in co-treatment, whereas all HP extracts significantly reduced the genotoxicity in the three protocols. Conclusion: These results suggested that HS was strongly involved in antimutagenic activity of HP extracts in co-treatment, but other components in HP extracts participated in this activity in pre- and post-treatment.
  1 1,761 15
Neuroprotective effects of Nigella sativa extracts during germination on central nervous system
Mohammad Hayatul Islam, Iffat Zareen Ahmad, Mohammad Tariq Salman
May 2015, 11(42):182-189
DOI:10.4103/0973-1296.157729  PMID:26109765
Background: Nigella sativa Linn. which has many acclaimed medicinal properties is an indigenous herbaceous plant and belongs to the Ranunculaceae family, which grows in countries bordering the Mediterranean Sea, Pakistan and India. Objective: This study was designed to investigate the effects of N. sativa seed extracts of different germination phases on the central nervous system (CNS) responses in experimental animals. Materials and Methods: Anxiolytic, locomotor activity of extracts (1 g/kg of body weight) was evaluated in both stressed and unstressed animal models and antiepileptic effect was evaluated by maximal electroshock seizure model keeping diazepam (20 mg/kg) as a positive control. Antidepressant effect was evaluated by forced swim test and tail suspension test keeping imipramine (15 mg/kg) as a positive control. Results: All tested extracts of N. sativa during different phases of germination (especially 5 th day germination phase) showed significant (P < 0.001) anxiolytic effect in comparison to control. Diazepam reduced locomotor activity in control (unstressed) rats but did not show affect in stressed rats while N. sativa extracts from germination phases significantly (P < 0.001) reduced locomotor activity in unstressed as well as stressed animals. All the extracts of N. sativa from different germination phases exhibited significant (P < 0.001) reduction in various phases of epileptic seizure on comparison with the reference standard (diazepam). During antidepressant test, N. sativa extracts exhibited a slight reduction in the immobility of rats. Conclusion: During germination, especially in 5 th day germination extract, N. sativa showed significant CNS depressant activity as compared to whole seeds that possibly may be due higher content of secondary metabolites produced during germination.
  1 3,703 15
Effects of the extract and glycoalkaloids of Solanum lycocarpum St. Hill on Giardia lamblia trophozoites
Gilmarcio Z Martins, Raquel R. D. Moreira, Cleopatra S Planeta, Adélia E Almeida, Jairo K Bastos, Lígia Salgueiro, Carlos Cavaleiro, Maria do Céu Sousa
May 2015, 11(42):161-165
DOI:10.4103/0973-1296.157721  PMID:26109762
Background: Solanum lycocarpum has great importance for food and medicinal traditional use. Recently, it was also evidenced that extracts of S. lycocarpum St. Hill (Solanaceae) and its glycoalkaloids, solamargine (Sg) and solasonine (Sn), are active against flagellated protozoa. Objective: The aim was to assess the effects of the extract of S. lycocarpum and its glycoalkaloids, Sn, and Sg, on Giardia lamblia trophozoites. Materials and Methods: A crude extract (96% ethanol) (EB) of fruits of S. lycocarpum was prepared and fractionated by partition with 40% ethanol and n-hexane: Ethyl acetate. Glycoalkaloids, Sn, and Sg were recognized in the ethanol fraction (EF) and further isolated by column chromatography. EB, EF, the isolated Sn and Sg and a mixture (1:1) of both glycoalkaloids were tested on cultures of G. lamblia trophozoites and macrophages. Results: EB, EF and glycoalkaloids of S. lycocarpum showed activity against Giardia (95.0 < Inhibitory concentration 50 [IC 50 ] ≤120.3 μg/mL). The mixture of glycoalkaloids (1:1) was more active (IC 50 = 13.23 μg/mL) than each one individually, suggesting a synergic effect. Moreover, the mixture is nontoxic to macrophage cells. Conclusion: Results are optimistic concerning the anti-Giardia potential of the mixture Sn + Sg. Further studies, in vitro and in vivo, will be required to consolidate the usefulness of the mixture of Sn + Sg in view of a new therapeutic strategy for giardiasis.
  1 1,697 14
Development of response surface methodology for optimization of extraction parameters and quantitative estimation of embelin from Embelia ribes Burm by high performance liquid chromatography
Md. Shamsir Alam, Zoheir A Damanhouri, Aftab Ahmad, Lubna Abidin, Mohd Amir, Mohd Aqil, Shah Alam Khan, Mohd Mujeeb
May 2015, 11(42):166-172
DOI:10.4103/0973-1296.157722  PMID:26109763
Background: Embelia ribes Burm is widely used medicinal plant for the treatment of different types of disorders in the Indian traditional systems of medicine. Objective: The present work was aimed to optimize the extraction parameters of embelin from E. ribes fruits and also to quantify embelin content in different extracts of the plant. Materials and Methods: Optimization of extraction parameters such as solvent: drug ratio, temperature and time were carried out by response surface methodology (RSM). Quantitative estimation of embelin in different extracts of E. ribes fruits was done through high performance liquid chromatography. Results: The optimal conditions determined for extraction of embelin through RSM were; extraction time (27.50 min), extraction temperature 45°C and solvent: drug ratio (8:1). Under the optimized conditions, the embelin yield (32.71%) was equitable to the expected yield (31.07%, P > 0.05). These results showed that the developed model is satisfactory and suitable for the extraction process of embelin. The analysis of variance showed a high goodness of model fit and the accomplishment of the RSM method for improving embelin extraction from the fruits of E. ribes. Conclusion: It is concluded that this may be a useful method for the extraction and quantitative estimation of embelin from the fruits of E. ribes.
  1 1,931 17
Pharmacophore modeling, in silico screening, molecular docking and molecular dynamics approaches for potential alpha-delta bungarotoxin-4 inhibitors discovery
R Barani Kumar, M Xavier Suresh, B Shanmuga Priya
May 2015, 11(42):19-28
DOI:10.4103/0973-1296.157670  PMID:26109766
Background: The alpha-delta bungartoxin-4 (α-δ-Bgt-4) is a potent neurotoxin produced by highly venomous snake species, Bungarus caeruleus, mainly targeting neuronal acetylcholine receptors (nAchRs) and producing adverse biological malfunctions leading to respiratory paralysis and mortality. Objective: In this study, we predicted the three-dimensional structure of α-δ-Bgt-4 using homology modeling and investigated the conformational changes and the key residues responsible for nAchRs inhibiting activity. Materials and Methods: From the selected plants, which are traditionally used for snake bites, the active compounds are taken and performed molecular interaction studies and also used for modern techniques like pharmacophore modeling and mapping and absorption, distribution, metabolism, elimination and toxicity analysis which may increase the possibility of success. Results: Moreover, 100's of drug-like compounds were retrieved and analyzed through computational virtual screening and allowed for pharmacokinetic profiling, molecular docking and dynamics simulation. Conclusion: Finally the top five drug-like compounds having competing level of inhibition toward α-δ-Bgt-4 toxin were suggested based on their interaction with α-δ-Bgt-4 toxin.
  - 2,347 17
Phytoecdysteroids of the East Asian Caryophyllaceae
Elena Novozhilova, Viacheslav Rybin, Petr Gorovoy, Irina Gavrilenko, Roman Doudkin
May 2015, 11(42):225-230
DOI:10.4103/0973-1296.157746  PMID:26109770
Background: Occurrence of integristerone A (1), 20-hydroxyecdysone (2), ecdysone (3), 2-deoxy-20-hydroxyecdysone (4) has been analyzed in 64 species of the East Asian Caryophyllaceae. Materials and Methods: Ecdysteroid content was determinate by high-performance liquid chromatography (HPLC). HPLC with a high-resolution mass spectrometry was performed on Shimadzu LCMS-IT-TOF (Japan) system equipped with a LC-20A Prominence liquid chromatograph, a photodiode array detector SPD-M20A and ion-trap/time-of-flight mass spectrometer. Results: New sources of phytoecdysteroids: Melandrium sachalinense and Melandrium firmum have been revealed. It is the 1 st time that two has been identified in M. sachalinense and M. firmum; 1 in the species: Lychnis fulgens, Silene repens, Silene foliosa, Silene stenophylla, Silene jenisseensis and M. sachalinense; 3 in Lychnis cognata; 4 in L. fulgens, S. stenophylla and S. jenisseensis (the tribe Lychnideae, the subfamily Caryophylloideae). Ecdysteroid-negative taxa are Spergularia rubra of the tribe Sperguleae; species of the genera Minuartia, Honckenya, Eremogone, Arenaria, Moehringia, Pseudostellaria, Fimbripetalum, Stellaria and Cerastium of the tribe Alsineae; Scleranthus annuus of the tribe Sclerantheae, as well as the East Asian representatives of the genera Gypsophila, Psammophiliela, Dianthus and Saponaria of the tribe Diantheae; Oberna and Agrostemma of the tribe Lychnideae. Conclusion: This investigation shows the most promising sources of ecdysteriods are species of genera Silene and Lychnis.
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Protective role of Kalpaamruthaa in type II diabetes mellitus-induced cardiovascular disease through the modulation of protease-activated receptor-1
Raja Latha, Palanivelu Shanthi, Panchanadham Sachdanandam
May 2015, 11(42):209-216
DOI:10.4103/0973-1296.157739  PMID:26109768
Background: Kalpaamruthaa (KA) is a formulatory herbal preparation has beneficial antioxidant, anti-apoptotic and anti-inflammatory properties against cardiovascular damage (CVD). Objective: The present study was undertaken to investigate the protective role of KA in type II diabetes mellitus-induced CVD through the modulation of protease-activated receptor-1 (PAR1). Materials and Methods: CVD was developed in 8 weeks after type II diabetes mellitus induction with high fat diet (2 weeks) and low dose of streptozotocin (2 × 35 mg/kg b.w. i.p. in 24 h interval). CVD-induced rats treated with KA (200 mg/kg b.w. in 0.5 ml of olive oil) orally for 4 weeks. Results: KA increased the activities of enzymatic antioxidants and the levels of non-enzymatic antioxidants in pancreas of CVD-induced rats. KA effectively reduced the lipid peroxides and carbonyl content in the pancreas of CVD-induced rats. KA reduced cellular damage by ameliorating the activities of marker enzymes in plasma, heart and liver. The protective nature of KA was further evidenced by histological observation in pancreas. Further, KA reduced CVD by decreasing the expression of PAR1 in heart. Conclusion: This study exhibits the defending role of KA in type II diabetes mellitus-induced CVD through altering PAR1.
  - 1,425 14
Antiproliferative efficacy of Tabernaemontana divaricata against HEP2 cell line and Vero cell line
Arvind Kumar, S Selvakumar
May 2015, 11(42):46-52
DOI:10.4103/0973-1296.157682  PMID:26109773
Background: Laryngeal cancer may also be called cancer of the larynx or laryngeal carcinoma. Conventional plants are a precious source of novel anticancer agents and are still in performance better role in health concern. The study was intended to estimation of the anticancer activity of the chloroformic extract of Tabernaemontana divaricata on the human epidermoid larynx carcinoma cell line (Hep 2). Materials and Methods: The aerial parts (leaves, stem, and flowers) of T. divaricata were tested for its inhibitory effect in 96 microplate formats against Hep 2 cell line. The anticancer activity of samples on Hep 2 and Vero was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and various enzymatic parameters like catalase, reduced glutathione (GSH), GSH peroxidase, and superoxide anion scavenging activity. Viable cells were determined by the absorbance at 540 nm. Measurements were performed, and the concentration required for a 50% inhibition of viability (IC50) was determined graphically. The effect of the samples on the proliferation of Hep 2 and Vero cells was expressed as the % cell viability. Results: The extract on Hep 2 cell line up to 7.8 μg/ml and that IC50 value on Hep 2 cell line was 112 μg whereas 94 μg for Vero cell line. Hence, T. divaricata has lesser significant action on Vero cell line. Conclusion: Medicinal plant drug discovery continues to provide new and important leads against various pharmacological targets including cancer. Our results clearly indicate the anticancer property of the medicinal plant T. divaricata against the human laryngeal carcinoma cell lines (Hep 2 cell line).
  - 2,035 16
Assay method for quality control and stability studies of a new anti-diabetic and anti-dyslipidemic flavone (S002-853)*
Arshi Naqvi, Anuja Pandey, Varsha Gupta, Richa Malasoni, Akansha Srivastava, Rishi Ranjan Pandey, M Satyanarayana, Ram Pratap, Anil Kumar Dwivedi
May 2015, 11(42):53-59
DOI:10.4103/0973-1296.157689  PMID:26109774
Background: Flavonoid-rich extract of the plant is long known for its anti-diabetic activities in traditional medicine. S002-853, a new flavone derivative synthesized by Central Drug Research Institute (CDRI) has been used for the present study. Objectives: The present study aimed at development of an assay method for quality control (QC) and stability studies of a new anti-diabetic and anti-dyslipidemic agent CDRI compound S002-853. Materials and Methods: A validated high-performance liquid chromatography analysis method for S002-853 was developed for in process QC and stability studies. The separation was achieved on a RP-C18 (25 cm × 0.4 cm, 5 μm, Phenomenex) at 240 nm with flow rate of 1.0 ml/min. This method was applied successfully in establishing forced degradation and drug-excipient testing protocols as per International Conference on Harmonization guidelines. Results: The result of estimation and stress testing studies indicated a high degree of selectivity of this method. S002-853 was most stable at pH 7 and under photolytic conditions. The temperature degradation pattern of S002-853 was found to follow the zero order degradation. Conclusion: The method described is easy and simple hence can be easily reproduced. This method can be very useful for bulk manufacture QC, and drug development process.
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Use of bergenin as an analytical marker for standardization of the polyherbal formulation containing Saxifraga ligulata
Hulikal Basavarajaiah Pushpalatha, Kumar Pramod, Rajendran Devanathan, Ramachandran Sundaram
May 2015, 11(42):60-65
DOI:10.4103/0973-1296.157690  PMID:26109776
Background: Bergenin is the principle constituent of the well-known medicinal plant Saxifraga ligulata. Bergenin has anti-inflammatory, antipyretic, antiviral, immunostimulant, antihyperglycemic, and antioxidant properties. In this study, the presence of bergenin in Saxifraga ligulata and the formulations was identified using high performance thin layer chromatograpy fingerprinting technique. Objective: To develop a novel quantitative method for the estimation of bergenin using high performance liquid chromatography. Materials and Methods: The compound was separated, characterised and quantified using authentic reference standard. The method was validated per ICH guidelines for the parameters of accuracy, precision, linearity, limit of detection, limit of quantification and robustness. Results: The method was found to be accurate, linear (r2 = 0.998) and precise (%RSD <2%). The limits of detection (0.001%) and quantification (0.002%) were found to be suitable for detection and quantification of bergenin in commercial formulations. Conclusion: The developed methods are suitable for the quality control applications of Saxifraga ligulata containing formulations.
  - 1,563 15
A novel high-performance liquid chromatography-electron spray ionization-mass spectrometry method for simultaneous determination of guggulsterones, piperine and gallic acid in Triphala guggulu
Ganesh Muguli, PR Rao Vadaparthi, B Ramesh, Vishakante Gowda, Rangesh Paramesh, Atul N Jadhav, K Suresh Babu
May 2015, 11(42):66-72
DOI:10.4103/0973-1296.157696  PMID:26109777
"Triphalaguggulu0" is an important Ayurvedic formulation comprising of Guggulu, that is, Commiphora wightii (Arn.) Bhandari as a base wherein powdered fruits of triphala, that is, Phyllanthus emblica L., Terminalia bellirica (Gaertn.) Roxb and Terminalia chebula Retz, along with powdered fruit of Piper longum L. are compounded. This polyherbal preparation has been strongly recommended in chronic inflammation, piles, and fistula. However, due to the complexity of compound formulation standardization of commercial products is challenging. In the present communication marker-based standardization of "Triphalaguggulu" preparation using gallic acid (for triphala), piperine (for P. longum L.) and guggulsterones (for guggulu) is reported. These compounds of diverse chemistry were successfully separated on a Waters HR-C18 column by isocratic elution with methanol and water (80:20 v/v) as mobile phase at the flow rate of 1.0 mL/min coupled with photodiode array detector. These optimal chromatographic conditions were used for simultaneous quantification of gallic acid, guggulsterones (E and Z) and piperine in commercial samples by high-performance liquid chromatography-electron spray ionization-mass spectrometry and method was validated as per ICH guidelines.
  - 2,171 15
New free radical scavenging neolignans from fruits of Piper attenuatum
S Divya Reddy, Bandi Siva, B Poornima, D Anand Kumar, Ashok K Tiwari, U Ramesh, K Suresh Babu
April-June 2015, 11(42):235-241
DOI:10.4103/0973-1296.153063  PMID:25829760
Objective: The aim was to study and identify free radicals scavenging and antihyperglycemic principles in fruit of Piper attenuatum. Materials and Methods: Bioassay guided identification of extracts possessing potent free radical scavenging activity, and isolation of compounds was done. Chloroform extract of P. attenuatum possessing potent radical scavenging activity was also evaluated for antihyperglycemic activity following oral glucose tolerance test in rats. Results: Nine neolignans namely, denudatin B (1), iso-4', 5'- dimethoxy-3, 4-methylenedioxy-2'-oxo-Δ 3',5',8' -8.1'- lignan (2), lancifolin D (3), denudatin A (4), wallichinin (5), piperenone (6), lancifolin C (7), 2-oxo-piperol B (8), piperkadsin A (9) and a crotepoxide (10) was identified in Chloroform extract of P. attenuatum. Neolignans (1-9) displayed potent 2, 2'- azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) radical and piperkadsin A (9) also displayed 1, 1-diphenyl-2-picrylhydrazyl radical scavenging activity. Analysis of structure-activity relationship revealed that presence of furan ring and methoxy groups is an important criterion to influence 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging potentials. Chloroform extract of P. attenuatum fruit could not display antihyperglycemic activity following oral glucose tolerance test in rats. Conclusion: Neolignans present in P. attenuatum fruits are potent free radical scavengers and this is the first report identifying these compounds and activities in this fruit.
  - 3,565 19
Simultaneous determination three phytosterol compounds, campesterol, stigmasterol and daucosterol in Artemisia apiacea by high performance liquid chromatography-diode array ultraviolet/visible detector
Jiwoo Lee, Jin Bae Weon, Bo-Ra Yun, Min Rye Eom, Choong Je Ma
April-June 2015, 11(42):297-303
DOI:10.4103/0973-1296.153082  PMID:25829768
Background: Artemisia apiacea is a traditional herbal medicine using treatment of eczema and jaundice in Eastern Asia, including China, Korea, and Japan. Objective: An accurate and sensitive analysis method using high performance liquid chromatography-diode array ultraviolet/visible detector and liquid chromatography-mass spectrometry for the simultaneous determination of three phytosterol compounds, campesterol, stigmasterol and daucosterol in A. apiacea was established. Materials and Methods: The analytes were separated on a Shiseido C 18 column (5 μm, 4.6 mm I.D. ×250 mm) with gradient elution of 0.1% trifluoroacetic acid and acetonitrile. The flow rate was 1 mL/min and detection wavelengths were set at 205 and 254 nm. Results: Validation of the method was performed to demonstrate its linearity, precision and accuracy. The calibration curves showed good linearity (R2 > 0.9994). The limits of detection and limits of quantification were within the ranges 0.55-7.07 μg/mL and 1.67-21.44 μg/mL, respectively. And, the relative standard deviations of intra- and inter-day precision were <2.93%. The recoveries were found to be in the range of 90.03-104.91%. Conclusion: The developed method has been successfully applied to the analysis for quality control of campesterol, stigmasterol and daucosterol in A. apiacea.
  - 3,713 16
A new method for identification of natural, artificial and in vitro cultured Calculus bovis using high-performance liquid chromatography-mass spectrometry
Yonggang Liu, Peng Tan, Shanshan Liu, Hang Shi, Xin Feng, Qun Ma
April-June 2015, 11(42):304-310
DOI:10.4103/0973-1296.153083  PMID:25829769
Objective: Calculus bovis have been widely used in Chinese herbology for the treatment of hyperpyrexia, convulsions, and epilepsy. Nowadays, due to the limited source and high market price, the substitutes, artificial and in vitro cultured Calculus bovis, are getting more and more commonly used. The adulteration phenomenon is serious. Therefore, it is crucial to establish a fast and simple method in discriminating the natural, artificial and in vitro cultured Calculus bovis. Bile acids, one of the main active constituents, are taken as an important indicator for evaluating the quality of Calculus bovis and the substitutes. Several techniques have been built to analyze bile acids in Calculus bovis. Whereas, as bile acids are with poor ultraviolet absorbance and high structural similarity, effective technology for identification and quality control is still lacking. Methods: In this study, high-performance liquid chromatography (HPLC) coupled with tandem mass spectrometry (LC/MS/MS) was applied in the analysis of bile acids, which effectively identified natural, artificial and in vitro cultured Calculus bovis and provide a new method for their quality control. Results: Natural, artificial and in vitro cultured Calculus bovis were differentiated by bile acids analysis. A new compound with protonated molecule at m/z 405 was found, which we called 3α, 12α-dihydroxy-7-oxo-5α-cholanic acid. This compound was discovered in in vitro cultured Calculus bovis, but almost not detected in natural and artificial Calculus bovis. A total of 13 constituents was identified. Among them, three bio-markers, including glycocholic acid, glycodeoxycholic acid and taurocholic acid (TCA) were detected in both natural and artificial Calculus bovis, but the density of TCA was different in two kinds of Calculus bovis. In addition, the characteristics of bile acids were illustrated. Conclusions: The HPLC coupled with tandem MS (LC/MS/MS) method was feasible, easy, rapid and accurate in identifying natural, artificial and in vitro cultured Calculus bovis.
  - 2,758 15
Comparative evaluation of standardized alcoholic, hydroalcoholic, and aqueous extracts of Phyllanthus maderaspatensis Linn. against galactosamine-induced hepatopathy in albino rats
UK Ilyas, Deepshikha P Katare, Vidhu Aeri
April-June 2015, 11(42):277-282
DOI:10.4103/0973-1296.153079  PMID:25829765
Background: Phyllanthus maderaspatensis species (Euphorbiaceae) has been used in folk medicine of many countries as a remedy against several pathological conditions including jaundice and hepatitis. This study is an attempt to evaluate hepatoprotective activity of P. maderaspatensis against galactosamine-induced toxicity and also investigation of polyphenols in each extract. Materials and Methods: The extraction of P. maderaspatensis as per Ayurveda was simultaneously standardized and quantified for biochemical markers viz., polyphenols: Kaempferol, quercetin, catechin, rutin, and ellagic acid by high-performance thin layer chromatography. Hepatotoxicity was induced albino adult rats by intraperitoneal injection of galactosamine (400 mg/kg). The quantified aqueous, hydroalcoholic and alcoholic extract of P. maderaspatensis (200 and 400 mg/kg body weight/day) were compared for evaluation of hepatoprotective potential, which were assessed in terms of reduction in histological damage, change in serum enzymes such as aspartate amino transaminase, alanine amino transaminase and alkaline phosphatase and increase thiobarbituric acid reactive substances. Results and Discussion: The hydroalcoholic extract was found to contain comparatively high amount of kaempferol, quercetin, catechin, rutin, and ellagic acid which are responsible for hepatoprotection. Antioxidant parameters such as glutathione, catalase, and superoxide dismutase activity in liver tissues were restored toward the normalization more significantly by the hydroalcoholic extract when compared with other extracts. The biochemical observations were supplemented with histopathological examination. Conclusion: The hydroalcoholic extract standardized with respect to known biomarkers may be considered as a potent extract against hepatotoxicity.
  - 2,714 14
Effects of Valerianae Radix et Rhizoma extract on psychological stress in mice
Jeong Suk Kim, Jeong Deok Ahn, Su-In Cho
April-June 2015, 11(42):381-388
DOI:10.4103/0973-1296.153093  PMID:25829779
Background: The aim of this study was to identify the effects of Valerianae Radix et Rhizoma water extract (VRe) originated from Valeriana fauriei Briquet on reducing psychological stress (PS) on mice. Objective: Mice were put under PS with communication box method: Restraining mice and forcing to see other mice underfoot shock stress. Materials and Methods: Measurements on plasma corticosterone, noradrenaline and lipid peroxidation, and elevated plus-maze (EPM) tests were carried out to determine the effect of VRe administration on physiological and behavioral responses of mice. Results: VRe showed anxiolytic effects in plasma corticosterone, noradrenaline, and EPM transfer latency levels, but it did not show any significant effects on the other indicators. Conclusion: V. fauriei, which has been used as a natural anxiolytic drug, exerts positive effects in the communication box induced PS in mice.
  - 2,489 15
Flavonoid profile, antioxidant and cytotoxic activity of different extracts from Algerian Rhamnus alaternus L. bark
Soulef Boussahel, Antonio Speciale, Saliha Dahamna, Yacine Amar, Irene Bonaccorsi, Francesco Cacciola, Francesco Cimino, Paola Donato, Guido Ferlazzo, Daoud Harzallah, Mariateresa Cristani
May 2015, 11(42):102-109
DOI:10.4103/0973-1296.157707  PMID:26109754
Background: Rhamnus alaternus (Rhamnaceae) L. has been traditionally used for treatment of many diseases. Objective: In this study, we determined the antioxidant/free radical scavenger properties, the flavonoid profile and the cytotoxicity of aqueous and methanolic extracts obtained by maceration from Algerian R. alaternus bark, like also of aqueous extract prepared by decoction according to the traditional method. This to estimate the usefulness of the drug traditional preparation and compare it with those made in the laboratory. Materials and Methods: The antioxidant activity of the extracts was evaluated using five different redox-based assays, all involving one redox reaction with the oxidant. High-performance liquid chromatography/diode array detection/electrospray ionization mass spectrometry analysis was used to identify and quantify the fl avonoids content. Cytotoxicity on human monocytic leukemia cells (U937) was also carried out. Results: All the extracts tested showed a good antioxidant/free radical scavenger activity and a similar flavonoid fingerprint. However, the methanolic one presented the best antioxidant activity that can be due to the highest flavonoid amount and significantly reduced the proliferation of leukemia cells. The results confirm that the extract prepared by decoction contains efficient antioxidant compounds and this justifies in part the therapeutic and preventive usefulness. Moreover, the methanolic extract exerted excellent cytotoxicity on U937 that could be attributed to kaempferol and rhamnocitrin glycosides.
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In vivo antitumor potential of Ipomoea pes-caprae on melanoma cancer
Ashish Manigauha, MD Kharya, N Ganesh
April-June 2015, 11(42):426-433
DOI:10.4103/0973-1296.153099  PMID:25829785
Background: The incidence of skin cancers is rising gradually. The treatment of melanoma is also necessary to prevent the spread of cancer to other body organs. Scientific literatures have not documented any evidence of the antitumor potential of Ipomoea pes-caprae on melanoma. Aim of the Study: Explore in vivo antitumor potential of I. pes-caprae on melanoma cancer. Materials and Methods: Petroleum ether (60°C-80°C), methanolic and aqueous extracts, and swaras prepared from the whole herb of I. pes-caprae were assessed for their antitumor activity. The extracts and swaras at doses of 25 and 50 mg/kg b. wt. were administered intraperitoneal along with chemo and radiotherapy for 40 days for exploring antitumor activity against melanoma cancer (B16F10) in male C57BL mice. The results obtained from tumor volume, and histopathological studies were compared with the control and dacarbazine used as a standard. Results: Antitumor effect of I. pes-caprae extracts and swaras on melanoma cancer was found to be significant (P < 0.01) compared to normal control. The tumor volume inhibition against tumor-bearing mice, although differed from each other, was concentration dependent. Administration of plant extracts and swaras from the day 1 since tumor inducted. The induction of tumor was found delayed by 10-15 days and the tumor volume on the day 40 was similar to the Dacarbazine treatment used as a standard. Conclusion: The results obtained from the tumor volume and histopathological studies clearly revealed the antitumor potential of I. pes-caprae on melanoma cancer.
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Bacopa monnieri promotes longevity in Caenorhabditis elegans under stress conditions
Suresh C Phulara, Virendra Shukla, Sudeep Tiwari, Rakesh Pandey
April-June 2015, 11(42):410-416
DOI:10.4103/0973-1296.153097  PMID:25829783
Background: Bacopa monnieri (L.) Pennell, commonly known as Brahmi is an important medicinal plant traditionally used as memory enhancer and antiepileptic agent. Objective: The present study investigated antioxidant and stress resistance potentials of B. monnieri aqueous extract (BMW) using Caenorhabditis elegans animal model system. Materials and Methods: The antioxidant activity of the BMW was measured using in vitro (DPPH, reducing power and total polyphenol content) and in vivo (DCF-DA assay) assays. The antistress potential of BMW (0.1, 0.01, and 0.001 mg/ml) was evaluated through thermal stress (37°C) and oxidative stress (10 mM paraquat) using C. elegans. Quantification of the HSP-16.2 level was done using CL2070 transgenic worms. Results: Present study reveals that BMW possess in vitro and in vivo antioxidant activities. BMW significantly enhanced stress tolerance and increased the mean lifespan of worms during thermal and oxidative stress, although it did not extend lifespan at 20°C and attenuated age dependent decline in physiological behaviors. Moreover, it was shown that BMW was able to up-regulate expression of stress associated gene hsp-16.2, which significantly (P < 0.001) extends the mean lifespan of worms under stress conditions. Conclusion: The study strongly suggests that BMW acts as an antistressor and potent reactive oxygen species scavenger which enhances the survival of the worms in different stress conditions.
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