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   2016| July  | Volume 12 | Issue 47  
    Online since September 30, 2016

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Sambar, an Indian dish prevents the development of dimethyl hydrazine-induced colon cancer: A preclinical study
Vutturu Ganga Prasad, Neetinkumar Reddy, Albi Francis, Pawan G Nayak, Anoop Kishore, Krishnadas Nandakumar, Mallikarjuna C Rao, Rekha Shenoy
July 2016, 12(47):441-445
DOI:10.4103/0973-1296.191454  PMID:27761072
Background: Colon cancer (CC) is the third commonly diagnosed cancer and the second leading cause of mortality in the US when compared to India where prevalence is less. Possible reason could be the vegetarian diet comprising spices used in curry powders. Researchers believe that 70% of the cases are associated with diet. Spices have inherited a rich tradition for their flavor and medicinal properties. Researchers have been oriented towards spices present in food items for their antitumorigenic properties. Objective: We investigated the effects of sambar as a preventive measure for 1,2-dimethyl hydrazine (DMH)-induced CC in Wistar albino rats. Materials and Methods: The animals were divided into three groups (n = 6) namely control, DMH, and sambar. At the end of the experimental period, the animals were killed using anesthesia and the colons and livers were examined. Results: All the treatment groups exhibited a significant change in the number of aberrant crypt foci (ACF). Sambar group showed a significant change in the colonic GSH when compared to both normal and DMH groups. A significant reduction in the liver GSH was noted in the sambar group. Only sambar group showed a significant change in the liver catalase levels when compared to DMH. There was a significant reduction in the colonic nitrite in the sambar-treated group; 2.94 ± 0.29 when compared to DMH control at 8.09 ± 1.32. On the contrary, a significant rise in the liver nitrite levels was observed in the sambar-treated rats. Conclusion: Sambar may prevent the risk of CC when consumed in dietary proportions. Abbreviations used: ACF: aberrant crypt foci, CC: colon cancer, DMH: 1,2-dimethyl hydrazine, GSH: glutathione, IL-6: Interleukin-6, TNF-α: Tumor necrosis factor-alpha.
  7,522 209 1
Anti-arthritic effects of total flavonoids from Juniperus sabina on complete freund's adjuvant induced arthritis in rats
Jun Zhao, Tao Liu, Fang Xu, Shuping You, Fang Xu, Chenyang Li, Zhengyi Gu
Jul-Sep 2016, 12(47):178-183
DOI:10.4103/0973-1296.186346  PMID:27601846
Context: Twigs and leaves of Juniperus sabina L. have been traditionally used as the medicinal herb in China for the treatment of many ailments including rheumatoid arthritis (RA). Aims: To confirm the therapeutic effect of total flavonoids from J. sabina (JSTF) on RA-induced by Complete Freund's Adjuvant (CFA) in rats. Settings and Design: Wistar rats (200 ± 20 g) were immunized by intradermal injection of 0.1 mL of CFA into the right hind metatarsal footpad. JSTF was administered orally at the dose of 125,250 and 500 mg/kg on 14 days after the induction of adjuvant arthritis. Tripterygium glycoside (20 mg/kg) was used as a positive control. Paw swelling, arthritic score, body weight loss, serum cytokines, inflammatory mediators, and histological change were measured. Results: We found that JSTF could ameliorate paw swelling of CFA rats, and significantly inhibit arthritic score (P < 0.05). The overproduction of tumor necrosis factor alpha and interleukin 1beta were remarkably suppressed in the serum of JSTF (125,500 mg/kg) treated rats (P < 0.05). Histopathological studies also showed a marked decrease of synovial inflammatory infiltration and synovial lining hyperplasia in the joints of JSTF-treated animals. Six flavonoids were isolated and from JSTF by various chromatographic methods and identified as follows: Catechin, quercitrin, isoquercitrin, isoscutellarein 7-O-β-D-xylopyranoside, isoscutellarein 7-O-β-D-xylopyranose-(1 → 3)-α-L-rhamnoside, and rutin. Conclusions: These results suggest the potential therapeutically effect of JSTF as an anti-arthritis agent toward CFA-induced arthritis in rats, and verified therapeutic applications of J. sabina on RA in folk medicine. SUMMARY
  • Twigs and leaves of Juniperus sabina L. have been traditionally used as the medicinal herb in China for the treatment of rheumatoid arthritis
  • JSTF could ameliorate paw swelling of CFA rats, and significantly inhibit arthritic score
  • Histopathological studies showed a marked decrease of synovial inflammatory infiltration and synovial lining hyperplasia in the joints of JSTF-reated animals
  • Six flavonoids were isolated and from JSTF including: Catechin, quercitrin, isoquercitrin, isoscutellarein 7-O-β-D-xylopyranoside, isoscutellarein 7-O-βD-xylopyranose.(1→3).αL-rhamnoside, and rutin.
Abbreviations used: JSTF: Total flavonoids from Juniperus sabina; CFA: Complete Freund's Adjuvant; TG: Tripterygium glycoside; TNF-α: Tumor necrosis factor alpha; IL-1β: Interleukin 1beta; IL-6: Interleukin 6; H and E: Hematoxylin and eosin.
  3,987 161 1
Induction, subculture cycle, and regeneration of callus in Safed musli (Chlorophytum borivilianum) using different types of phytohormones
Jaafar Juju Nakasha, Uma Rani Sinniah, Nurashikin Kemat, Kumara Swamy Mallappa
July 2016, 12(47):460-464
DOI:10.4103/0973-1296.191457  PMID:27761075
Background: Chlorophytum borivilianum is an industrially valued medicinal crop. Propagation through seeds is not feasible because of low germination percentage and long dormancy period. Therefore, callus culture and plant regeneration can be an alternative to improve this crop production. Also, callus can serve as an alternative source of bioactive compounds. Objective: To evaluate the effect of different phytohormones on callus induction, subculture cycle, and regeneration studies of callus in C. borivilianum. Materials and Methods: Young shoot buds of C. borivilianum were inoculated on Murashige and Skoog medium fortified with 3% sucrose and different concentrations (0, 1, 5, 10, and 15 mg/L) of either naphthalene acetic acid or 2,4-dichlorophenoxyacetic acid or indole-3-acetic acid and callus induction was evaluated up to four subcultures cycles. Shoot regeneration from callus was studied on Murashige and Skoog media fortified with 6-benzylaminopurine andkinetin or thidiazuron at varied levels (0, 0.5, 1, 2, and 3 mg/L). Microshoots were rooted on Murashige and Skoog media supplemented with 1.0 mg/L indole-3-butyric acid and plantlets were acclimatized before transferred to the natural conditions. Results: Callus induction was better evidenced on Murashige and Skoog media containing 5 mg/L 2,4-dichlorophenoxyacetic acid up to fourth subculture. Callus differentiated into shoots on Murashige and Skoog media fortified with 6-benzylaminopurine or kinetin, whereas thidiazuron completely failed to regenerate shoots. Furthermore, microshoots rooted on 1.0 mg/L indole-3-butyric acid containing Murashige and Skoog media. The rooted plantlets were successfully acclimatized and established in soil with 88.3% survivability. Conclusion: The type of auxins played an important role in inducing callus tissue from shoot bud explants of Safed musli. In future, this in vitro protocol could benefit in crop improvement programs and serve as a new source of bioactive compounds from Safed musli callus tissue for various therapeutic applications. Abbreviations used: MS: Murashige and Skoog, NAA: naphthalene acetic acid, 2,4-D: 2,4-dichlorophenoxyacetic acid, IAA: indole-3-acetic acid, BAP: 6-benzylaminopurine, Kn: Kinetin, TDZ: thidiazuron, IBA: indole-3-butyric acid, RCBD: Randomized Complete Block Design, DMRT: Duncan's Multiple Range Test
  3,521 343 -
In vitro antioxidant and enzymatic approaches to evaluate neuroprotector potential of Blechnum extracts without cytotoxicity to human stem cells
Juliana Maria de Mello Andrade, Renata Biegelmeyer, Roger Remy Dresch, Natasha Maurmann, Patrícia Pranke, Amélia T Henriques
Jul-Sep 2016, 12(47):171-177
DOI:10.4103/0973-1296.186349  PMID:27601845
Background: Investigation of selected plant extracts on multi-targets related to neurodegeneration, such as monoamine oxidases (MAO), cholinesterase enzymes, and antioxidant activities (AOA) is a useful tool for identification of new scaffolds. Objective: This work investigated biological effects of three Blechnum methanol extracts from Brazil and chemical profile of the most active sample. Materials and Methods: AOA included scavenging of hydroxyl and nitric oxide radicals, also lipid peroxidation inhibition. Enzymatic modulation of Blechnum binervatum, Blechnum brasiliense, and Blechnum occidentale extracts on MAO and cholinesterases was conducted. Moreover, total phenol content was performed with all samples, and high-performance liquid chromatography-diode array detection mass spectrometry HPLC-DAD-MS analysis was carried out with B. brasiliense. Possible toxic effects were evaluated on Wistar rats polymorphonuclear cells (PMN) and human stem cells. Results: B. brasiliense extract presented the highest phenolic amount (9.25 g gallic acid equivalent/100 g extract) and lowest IC50values (112.3 ± 2.61 and 176.1 ± 1.19 μg/mL) against hydroxyl radicals and on lipid peroxidation, respectively, showing strong AO effects. On nitric oxide assay and cholinesterase inhibition, all extracts were considered inactive. MAO-A selective action was evidenced, being B. brasiliense powerful against this enzyme (IC50: 72.7 μg/mL), followed by B. occidentale and B. binervatum (IC50: 130.85 and 165.2 μg/mL). No cytotoxic effects were observed on PMN and human stem cells treated with Blechnum extracts. HPLC-DAD-MS analysis of B. brasiliense allowed the identification of chlorogenic and rosmarinic acids. Conclusion: Our results especially highlight B. brasiliense, with pronounced phenols content and strong effects on selected targets related to neurodegeneration, being characterized as a natural safe source of bioactive hydroxycinnamic acids. SUMMARY
  • Blechnum crude extracts showed high phenolic amounts and valuable IC50 values on targets related with neurodegenerative disorders
  • Blechnum brasiliense was the most active sample, with strong radical scavenging and lipid peroxidation inhibition, also with monoamine oxidases: A selective modulation
  • No cytotoxic effects were observed on polymorphonuclear cells rat cells and human stem cells treated with Blechnum extracts
  • High-performance liquid chromatography-diode array detection-mass spectrometry analysis of Blechnum brasiliense allowed the identification of hydroxycinnamic derivatives: Chlorogenic and rosmarinic acids.
Abbreviations used: IC50: half maximal inhibitory concentration; MAO: monoamine oxidase; MAO-A: monoamine oxidase isoform A; MAO-B: monoamine oxidase isoform B; HO: hydroxyl radical.
  3,753 106 4
Assessment of In vitro antibacterial activity and cytotoxicity effect of Nigella sativa oil
Ayse Ruveyda Ugur, Hatice Turk Dagi, Bahadir Ozturk, Gulsum Tekin, Duygu Findik
July 2016, 12(47):471-474
DOI:10.4103/0973-1296.191459  PMID:27761077
Background: Methicillin resistance is a serious health concern since it has spread among Staphylococcus aureus and coagulase-negative Staphylococci (CoNS) that are frequent community and nosocomial pathogens worldwide. Methicillin-resistant strains are often resistant to other classes of antibiotics, making their treatment difficult. Nigella sativa oil is known to be active against Gram-positive cocci, yet its in vitro cytotoxicity is rarely investigated, is a proper and powerful candidate for treatment of methicillin-resistant isolates. Objectives: The aim of this study is to evaluate the in vitro antibacterial activity and cytotoxicity effect of N. sativa oil. Materials and Methods: The minimal inhibitory concentrations (MICs) of N. sativa oil were determined by broth microdilution method against four different American Type Culture Collection strains, 45 clinical isolates of methicillin-resistant S. aureus (MRSA), and 77 methicillin-resistant CoNS (MRCoNS). The effects of different dilutions (0.25 μg/mL, 0.5 μg/mL, and 1 μg/mL) of N. sativa oil on the proliferation of gingival fibroblasts were evaluated. Results: The MIC values of N. sativa oil against clinical isolates of Staphylococci were between <0.25 μg/mL and 1.0 μg/mL. Compared to the control group, there was no cytotoxic effect on the proliferation of the gingival fibroblasts. Conclusion: In the present study, the oil of N. sativa was very active against MRSA and MRCoNS and had no in vitro cytotoxicity at relevant concentrations. These findings emphasize that there is a requirement for further clinical trials on N. sativa oil for "safe" medical management of infections caused by methicillin-resistant Staphylococci. Abbreviation used: ATCC: American Type Culture Collection; CLSI: Clinical and Laboratory Standards Institute; CoNS: Coagulase-negative Staphylococci; DMEM: Dulbecco's modified Eagle's medium; DMSO: Dimethyl sulfoxide; FBS: Fetal bovine serum; HGF: Human gingival fi broblast; MIC: Minimal inhibitory concentration; MRCoNS: Methicillin-resistant CoNS;MRSA: Methicillin-resistant S. aureus
  3,526 294 -
Protective effect of baicalin against experimental colitis via suppression of oxidant stress and apoptosis
Jun Yao, Xu Cao, Ru Zhang, Ying-xue Li, Zheng-lei Xu, Ding-guo Zhang, Li-sheng Wang, Jian-yao Wang
Jul-Sep 2016, 12(47):225-234
DOI:10.4103/0973-1296.186342  PMID:27601854
Background: Baicalin is a bioactive ingredient extracted from the root of Scutellariae radix, which is used to treat ulcerative colitis (UC). Objective: We investigated the activity of baicalin on lipopolysaccharide-stimulated RAW264.7 cells and 2,4,6-trinitrobenzene sulfonic acid-induced rats, including the attenuation of oxidant stress and apoptosis. Materials and Methods: The severity of colitis was assessed by disease activity index. The activities of catalase (CAT), glutathione peroxidase (GSH-PX), superoxide dismutase (SOD), and the content of malondialdehyde (MDA) were determined by their corresponding kits. The terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) was performed to study whether experimental colitis was associated with intestinal epithelial cell (IEC) apoptosis and the effect of baicalin on IEC apoptosis. Western blot analysis and immunocytochemistry assay were applied to determine the protein expressions. The reactive oxygen species (ROS) level in the colon of UC rats treated with baicalin was determined by ROS assay kit. Results: Baicalin remarkably upregulated the activities of CAT, GSH-PX, and SOD and decreased the content of MDA in a dose-dependent manner in vitro and in vivo. The TUNEL-positive cells in rats treated baicalin were remarkably reduced. Both Western blot analysis and immunocytochemistry assay indicated that baicalin significantly decreased the expressions of transforming growth factor beta-1, Bax protein and upregulated the expression of Bcl-2 protein. In addition, the expressions of total and cleaved caspase-3, total and cleaved caspase-9 protein, Fas, and FasL in vitro were downregulated by the treatment with baicalin. Baicalin of different doses reduced the generation of ROS in UC rats. Conclusion: Taken together, these evidences provide scientific basics for the application of baicalin in the treatment of UC and suggest that baicalin exerts its effect via suppression of oxidant stress and apoptosis. SUMMARY
  • Baicalin remarkably upregulated the activities of catalase, glutathione peroxidase, and superoxide dismutase and decreased the content of MDA, both in vivo and in vitro
  • The terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling-positive cells in rats treated baicalin remarkably reduced in a concentration-dependent manner
  • Western blot analysis and immunocytochemistry assay indicated that baicalin significantly decreased the expressions of transforming growth factor beta-1, Bax protein, and upregulated the expression of Bcl-2 protein
  • The expressions of total and cleaved caspase-3, total and cleaved caspase-9 protein, Fas, and FasL in vitro were downregulated by the treatment with baicalin.
Abbreviations used: UC: Ulcerative colitis, LPS: Lipopolysaccharide, TNBS: 2,4,6-trinitrobenzene sulfonic acid, DAI: Disease activity index, CAT: Catalase, GSH-PX: Glutathione peroxidase, SOD: Superoxide dismutase, MDA: Malondialdehyde, TUNEL: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling, ROS: Reactive oxygen species, IEC: Intestinal epithelial cell, SD: Sprague-Dawley, HE: H and E, DNTB: 5,5'-dithiobis-2-nitrobenzoic acid, TBA: Thiobarbituric acid, TBARS: Thiobarbituric acid-reactive substances, S.D: Standard deviation, and PBS: Phosphate-buffered saline.
  3,527 182 2
Antioxidant activity of water-soluble polysaccharides from Brasenia schreberi
Huiwen Xiao, Xueru Cai, Yijun Fan, Aoxue Luo
Jul-Sep 2016, 12(47):193-197
DOI:10.4103/0973-1296.186343  PMID:27601849
Objective: In order to investigate the antioxidant activities of polysaccharides (BPL-1 and BPL-2), one of the most important functional constituents in Brasenia schreberi was isolated from the external mucilage of B. schreberi (BPL-1) and the plant in vivo (BPL-2). This paper examines the relationship between the content of sulfuric radicals and uronic acid in BPL and the antioxidant activity of BPL. Materials and Methods: The free radicals, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+) and 1,1-diphnyl-2-picrylhydrazyl (DPPH-), were used to determine the antioxidant activity of BPL. The Fourier-transform infrared spectroscopy of BPL-1 and BPL-2 revealed typical characteristics of polysaccharides. Results: The two sample types had different contents. This was proved by their different adsorption peak intensities. The IC50 values of BPL-1 (31.189 mg/ml) and BPL-2 (1.863 mg/ml) showed significant DPPH radical scavenging activity. Based on the quantification of ABTS radical scavenging, the IC50 value of BPL-1 (5.460 mg/ml) was higher than that of BPL-2 (0.239 mg/ml). Therefore, in terms of the reducing power, the IC50 value of BPL-1 was too high to determine, and the IC50 value of BPL-2 was found to be 50.557 mg/ml. Hence, the antioxidant activity and total reducing power were high, and they were greater in BPL-2 than in BPL-1. In addition, BPL-2 was found to have more sulfuric radicals and uronic acid than BPL-1. Conclusion: The contents of sulfuric radicals and uronic acid are significantly correlated to the antioxidant activity and reducing power of BPL; the more sulfuric radicals and uronic acid, the more antioxidant activity and reducing power BPL has. SUMMARY
  • The water-soluble crude polysaccharides obtained from the external mucilage and the Brasenia schreberi plant in vivo were confirmed to have high contents of sulfuric radicals and uronic acid
  • Both BPL-1 and BPL-2 exhibited antioxidative activity and reducing power, and their antioxidative activity gradually improved with increasing concentrations
  • The content of sulfuric radicals and uronic acid in BPL-1 and BPL-2 might explain their high antioxidant activity.
Abbreviations used: BPL-1:Polysaccharide were isolated from the external mucilage of B. Schreberi; BPL-2: Polysaccharide were isolated from the plant in vivo of B. schreberi; BPL:Polysaccharide were isolated from B. Schreberi.
  3,510 186 -
Elucidation of molecular mechanism(s) of cognition enhancing activity of Bacomind® : A standardized extract of bacopa monnieri
Shekhar Dethe, M Deepak, Amit Agarwal
July 2016, 12(47):482-487
DOI:10.4103/0973-1296.191464  PMID:27761079
Background: Bacopa monnieri (L.) Wettst., commonly known as Brahmi, is renowned in Indian traditional system for its potent memory enhancing activity, which has been validated by various scientific studies. Objective: The objective of this study was to understand the molecular mechanism of memory enhancing activity of BacoMind® (BM), a standardized extract of B. monnieri. Materials and Methods: BM was screened in vitro in a panel of cell-free and receptor-transfected cell assays. The purified enzymes/membrane homogenates/cells were incubated with substrate/standard ligand in the absence or presence of the test compound. The IC 50 values and EC 50 values were determined by nonlinear regression analysis of the concentration-response curves generated with mean replicate values using Hill equation curve fitting. Results: BM was found to inhibit three enzymes; Catechol-O-methyl transferase (COMT), Prolyl endopeptidase (PEP), and Poly (ADP-ribose) polymerase (PARP). It also had an antagonistic effect on serotonin 6 and 2A (5-HT 6 and 5-HT 2A ) receptors , known to influence the different neurological pathways, associated with memory and learning disorders, age-associated memory impairment. Conclusion: BM was found to inhibit three enzymes namely, Catechol-O-methyl transferase (COMT), Prolyl endopeptidase (PEP), and Poly (ADP-ribose) polymerase (PARP). It also exhibited an antagonistic effect on 5-HT 6 and 5-HT 2A receptors. Abbreviations used: HTRF: Homogenous time resolved fluorescence, cAMP: Cyclic adenosine monophosphate, CHO : Chinese hamster ovary, RFU : Relative fluorescence unit, pNP: Para nitro phenol, AMC: 7-amino-4-methylcoumarin, ELISA: Enzyme linked immunosorbent assay, Z-Pro-Pro-CHO: Z-prolyl-prolinal, HEK: Human embryonic kidney, TE: Trolox equivalent.
  3,393 286 -
Antioxidant and anti-inflammatory properties of Algerian Thymelaea microphylla coss. and dur. extracts
Khadidja Dehimi, Antonio Speciale, Antonina Saija, Saliha Dahamna, Roberto Raciti, Francesco Cimino, Mariateresa Cristani
Jul-Sep 2016, 12(47):203-210
DOI:10.4103/0973-1296.186345  PMID:27601851
Background: Thymelaea microphylla Coss. et Dur. (Thymelaeaceae) (TM) is a rare medicinal plant endemic to Algeria. Leaves decoction is used in folk medicine for anticancer, anti-inflammatory, and antidiabetic properties. Objective: Herein, the antioxidant and anti-inflammatory properties of different extracts from leaves and flowers of Algerian TM were evaluated. Materials and Methods: The study was carried out by in vitro cell-free assays (antioxidant/radical properties), ex vivo experiments (inhibition of prostaglandin E2 and thromboxane B2 release in human whole blood) and in vitro experiments on cell systems (cytotoxicity on peripheral blood mononuclear cells, and protective effects on human vein endothelial cells exposed to TNF-α). Results: The acetone TM extract showed significant antioxidant properties and excellent anti-inflammatory and cyclooxygenase-inhibitory activity, together with lack of toxicity on normal human blood cells; furthermore, it was able to protect endothelial cells against dysfunction induced by TNF-α, as shown by decrease in cell death, e-selectin expression and leukocyte adhesion. Conclusion: On these bases, TM leaves and flowers appear to be a good source of bioactive compounds with significant antioxidant and antiinflammatory capability, and potentially effective in prevention and treatment of pathological conditions related to oxidative stress and inflammation, such as endothelial dysfunction. SUMMARY
  • Thymelaea microphylla leaves and flowers are a good source of bioactive compounds with significant antioxidant/free radical scavenger and anti-inflammatory activity.
  • The acetone extract from leaves and flowers of Algerian Thymelaea microphylla possesses excellent cyclooxygenase-inhibitory activity.
  • This extract is able to protect against endothelial dysfunction, an early event in development of atherosclerosis and vascular diseases.
Abbreviations used: TM: Thymelaea microphylla; BCB: Beta-carotene bleaching; AcE: Acetone extract; PGE2: Prostaglandin E2; TxB2: Thromboxane B2; FL: Fluorescein; Cat: Catechin; DPPH: 2,2-diphenyl-1-picrylhydrazyl; ABTS: 2,2'-azinobis-(3-ethyl-benzothiazolin-6-sulfonic acid)+; Que: Quercetin; ORAC: Oxygen radical absorbance capacity; AAPH: 2,2'-azobis (2-methylpropionamidine)dihydrochloride; PMS/NADH: Phenazine methosulfate/nicotinamide adenine dinucleotide; HUVECs: Human umbilical vein endothelial cells. Antonio Speciale
  3,357 124 2
Inhibitory and cytotoxic activities of chrysin on human breast adenocarcinoma cells by induction of apoptosis
Saeed Samarghandian, Mohsen Azimi-Nezhad, Abasalt Borji, Malihe Hasanzadeh, Farahzad Jabbari, Tahereh Farkhondeh, Mohammad Samini
July 2016, 12(47):436-440
DOI:10.4103/0973-1296.191453  PMID:27761071
Objectives: Chrysin, an active natural bioflavonoid found in honey and many plant extracts, was first known for its antioxidant and anti-inflammatory effects. The fact that antioxidants have several inhibitory effects against different diseases, such as cancer, led to search for food rich in antioxidants. In this study, we investigated the antiproliferative and apoptotic effects of chrysin on the cultured human breast cancer cells (MCF-7). Materials and Methods: Cells were cultured in Roswell Park Memorial Institute medium and treated with different chrysin concentrations for three consecutive days. Cell viability was quantitated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The percentage of apoptotic cells was determined by flow cytometry using Annexin V-fluorescein isothiocyanate. Results: The MTT assay showed that chrysin had an antiproliferative effect on MCF-7 cells in a dose- and time-dependent manner. The 50% cell growth inhibition values for chrysin against MCF-7 cells were 19.5 and 9.2 μM after 48 and 72 h, respectively. Chrysin induced apoptosis in MCF-7 cells as determined by flow cytometry. Chrysin inhibits the growth of the breast cancer cells by inducing cancer cell apoptosis which may, in part, explain its anticancer activity. Conclusion: This study shows that chrysin could also be considered as a promising chemotherapeutic agent and anticancer activity in treatment of the breast cancer cells in future. Abbreviations used: Human breast cancer cells (MCF-7), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), phosphate-buffered saline (PBS), normal fi broblast mouse (L929).
  3,220 210 1
New phytochemical constituent and bioactivities of Horwoodia dicksoniae and Rumex cyprius
Mohammed F Abdelwahab, Sibghatullah Sangi, Hussam H Arafat, Ehab A Ragab
Jul-Sep 2016, 12(47):165-170
DOI:10.4103/0973-1296.186348  PMID:27601844
Background: Many plants growing in Saudi Arabia are used in folk medicine for treatment of several diseases. Objective: Information of the chemical constituents and biological activities of plants is desirable for the discovery of therapeutic agents and discovering the actual value of folkloric remedies. Materials and Methods: The compounds were isolated and purified using silica gel column chromatography and preparative high-performance liquid chromatography-diode array detector (HPLC-DAD) Method. The alcoholic extracts of these plants were evaluated for biological activities. Results: Isolation and characterization of 1-feruloyl-β-D-glucopyranoside (1) as well as new secondary metabolite tryptophan methyl ester (2) were isolated for the 1st time from the Horwoodia dicksoniae. The three flavones were isolated from Rumex cyprius identified as isoorientin (3), vitexin (4), and Cynarosid (5). The structures of these compounds were characterized by nuclear magnetic resonance and mass spectrometry analysis and comparing with literature. The compounds were isolated and purified using silica gel column chromatography and preparative HPLC-DAD Method. The alcoholic extracts of these plants were evaluated for antimicrobial activities against two Gram-positive bacteria, two Gram-negative bacteria, and four pathogenic fungi. Both plants showed good activities against Syncephalastrum racemosum and Streptococcus pneumoniae with minimal inhibitory concentrations (MICs) 0.98 and 1.95 μg/mL, respectively. H. dicksoniae showed good activity against Aspergillus fumigates with an MIC 1.95 μg/mL. The two extracts showed also effective free radical scavenging activities in the 1,1-diphenyl-2-picrylhydrazyl assay. H. dicksoniae exhibited remarkable cytotoxic activity against Human breast cancer mammary cancer cells-7, Human liver cancer human hepatoma carcinoma cells-2, and human lung carcinoma (A-549) cell lines. Conclusions: It was suggested that further work should be carried out to isolate, purify, and characterize the active constituents responsible for the activity of these plants. SUMMARY
  • New secondary metabolite Tryptophane methyl ester as well as 1-feruloyl-β-D-glucopyranoside were isolated for the first time from the HD.
  • Isoorientin, vitexin and Cynarosid were isolated from RC.
  • HD exhibited good activity against Aspergillus fumigates with an MIC 1.95 μg mL-1.
  • HD showed significant cytotoxic activity against Human breast cancer (MCF-7), Human liver cancer (HepG-2) and Human lung carcinoma (A-549) cell lines.
  3,302 87 -
Nanostructured lipid carriers loaded with baicalin: An efficient carrier for enhanced antidiabetic effects
Feng Shi, Zheng Wei, Yingying Zhao, Ximing Xu
Jul-Sep 2016, 12(47):198-202
DOI:10.4103/0973-1296.186347  PMID:27601850
Context: Recent studies have demonstrated that baicalin has antihyperglycemic effects by inhibiting lipid peroxidation. Baicalin is low hydrophilic and poorly absorbed after oral administration. Thus, a suitable formulation is highly desired to overcome the disadvantages of baicalin. Objective: The objective of this work was to prepare baicalin-loaded nanostructured lipid carriers (B-NLCs) for enhanced antidiabetic effects. Materials and Methods: B-NLCs were prepared by high-pressure homogenization method using Precirol as the solid lipid and Miglyol as the liquid lipid. The properties of the NLCs, such as particle size, zeta potential (ZP), and drug encapsulation efficiency (EE), were investigated. The morphology of NLCs was observed by transmission electron microscopy. In addition, drug release and antidiabetic activity were also studied. Results: The results revealed that B-NLCs particles were uniformly in the nanosize range and of spherical morphology with a mean size of 92 ± 3.1 nm, a ZP of −31.35 ± 3.08 mV, and an EE of 85.29 ± 3.42%. Baicalin was released from NLCs in a sustained manner. In addition, B-NLCs showed a significantly higher antidiabetic efficacy compared with baicalin. Conclusion: B-NLCs described in this study are well-suited for the delivery of baicalin. SUMMARY
  • Currently, herbal medicines have attracted increasing attention as a complementary approach for type 2 diabetes
  • Baicalin has antihyperglycemic effects by inhibiting lipid peroxidation
  • A suitable formulation is highly desired to overcome the disadvantages (poor solubility and low bioavailability) of baicalin
  • Nanostructured lipid carriers could enhance the antidiabetic effects of baicalin.
Abbreviations used: B-NLCs: Baicalin-Loaded Nanostructured Lipid Carriers, B-SUS: Baicalin Water Suspension, EE: Encapsulation Efficiency, FBG: Fasting Blood Glucose, HbAlc: Glycosylated Hemoglobin, HPLC: High-performance Liquid Chromatography; NLCs: Nanostructured Lipid Carriers, PI: Polydispersity Index, SD: Sprague-Dawley, SLNs: Solid lipid nanoparticles, STZ: Streptozotocin, TC: Total cholesterol, TEM: Transmission Electron Microscope, TG: Total Triglyceride, ZP: Zeta Potential.
  3,151 200 1
Antifatigue effects of ethanol extracts and polysaccharides isolated from Abelmoschus esculentus
Yu-Xian Li, Zhong-Han Yang, Yin Lin, Wei Han, Shan-Shan Jia, Ke Yuan
Jul-Sep 2016, 12(47):219-224
DOI:10.4103/0973-1296.186341  PMID:27601853
Background: The aim of this study is to determine the antifatigue active fraction from Abelmoschus esculentus. The in vivo antifatigue effects of ethanol extracts and polysaccharides from A. esculentus fruit have been determined. The polysaccharides of A. esculentus were determined as the best effective fractions of antifatigue effects. Materials and Methods: About 360 Kunming male mice were randomly divided into nine subgroups: normal control subgroup, model subgroup, positive subgroup and the ethanol extracts of A. esculentus with high dose (3.2 g/kg) subgroup, medium dose (1.6 g/kg) subgroup and low dose (0.8 g/kg) subgroup, the polysaccharides of high dose (3.2 g/kg) subgroup, medium dose (1.6 g/kg) subgroup, and the low dose (0.8 g/kg) subgroup. The antifatigue effects of ethanol extracts and polysaccharides form A. esculentus were measured by comparing body weight, food intake, swimming time, liver glycogen, serum urea, blood lactic acid as well as visceral parameter in mice. Results: Compared with the model subgroup, other subgroups significantly prolonged swimming time, and high dose polysaccharides administration was the most effective (P < 0.01). High dose polysaccharides significantly increased liver glycogen, serum lactic acid, and serum urea (P < 0.01) in mice. In contrast with model group, the high dose polysaccharides administration could also significantly elevated the parameters of testicles and epididymis (P < 0.01). The study established that the ethanol extracts and polysaccharides of A. esculentus both have antifatigue effects. Conclusions: The results demonstrated that both the ethanol extracts and polysaccharides of A. esculentus have antifatigue effects. The high dosage polysaccharides have significant antifatigue properties. The results will provide the basis for further development and utilization of this plant. SUMMARY
  1. The high dosage polysaccharides have restoration ability on kidney yang deficiency mice.
  2. The high dosage polysaccharides have significant effects of relieving body fatigue of mice.
  3. The polysaccharide of Abelmoschus esculentus showed better antifatigue effects than the ethanol extracts.
Abbreviations used: A. esculentus: Abelmoschus esculentus, BUN: Blood urine nitrogen, LD: Lactic Acid dehydrogenase, AE: Abelmoschus esculentus ethanol extracts, AP: Abelmoschus esculentus polysaccharides, LAC: Lactic acid content.
  3,194 122 -
In vitro accumulation of polyphenols in tea callus derived from anther
Naga Pavan Kumar Chevala, Naga Thirumalesh Chevala, Kirubakaran Dhanakodi, Rama Rao Nadendla, Chandrashekara Krishnappa Nagarathna
July 2016, 12(47):400-406
DOI:10.4103/0973-1296.191442  PMID:27761066
Background: Tea is an economic important crop with high medicinal value due to rich polyphenols content. In the present research we studied the accumulation of polyphenols of in vitro regenerated callus from anthers. Objective: Callus induction of tea anthers and in vitro accumulation of phenolic compounds from the anther-derived callus. Materials and Methods: Standardization of callus induction for tea anthers. In vitro generated callus was screened for in vivo accumulation of catechins and its isomers were screened by FC reagent staining technique. The methanol extract of dry and green callus obtained were estimated qualitatively by Fourier transform infrared spectroscopy (FTIR)-alternative total reflection (ATR) and quantitatively by HPLC method. Results: Anthers inoculated on half strength MS media fortified with 2,4-dichloro acetic acid (2 mg/L), Kn (1 mg/L), and BAP (1 mg/L) induced callus under photoperiod of 9:15 h light. The in vivo histochemical studies revealed the accumulation of polyphenols in the callus. The in vitro generated fresh and dry callus were used for extraction and screened for accumulated polyphenols [galic acid, (+)-catechin (C), (−)-epicatechin, (−)-epigallocatechin, (−)-epigallocatechin gallate, (−)-gallocatechins, (−)-epicatechin gallate] were estimated qualitatively by FTIR-ATR method and quantitatively by HPLC method. Conclusion: The FC staining technique used here helps in localization of polyphenol compounds accumulation in the tissues by instant microscopic studies. The study have scope in large-scale isolation of various medicinally important flavonol by using anther culture. Abbreviations used: HPLC: high pressure liquid chromatography; FTIR: Fourier transform infrared spectroscopy; 2,4-D: 2,4-dichloro acetic acid; BAP: N 6 -benzyl amino purine; kn: kinetin
  2,898 156 -
Free radical scavenging, α-glucosidase inhibitory and anti-inflammatory constituents from Indian sedges, Cyperus scariosus R.Br and Cyperus rotundus L.
Lavanya Kakarla, Suresh Babu Katragadda, Ashok K Tiwari, K Srigiridhar Kotamraju, K Madhusudana, D Anand Kumar, Mahendran Botlagunta
July 2016, 12(47):488-496
DOI:10.4103/0973-1296.191467  PMID:27761080
Background: Cyperus scariosus R. Br and Cyperus rotundus L are widely used in ayurvedic preparation for the treatment of diabetes and other diseases. The early literature ,so far, does not indicate the presence of any bioactive principle isolated from these plants. Objective: To identify free radical scavenging, anti-diabetic and anti- inflammatory principles from these two species. Materials and Methods: The bioassay guided fractionation and isolation of active constituents was done by chromatographic techniques. They also evaluated their anti-oxidant activity by DPPH and ABTS. The anti-diabetic activity was screened by α- glucosidase and α- amylase assays.Also, the further evaluation of in vitro anti-inflammatory activity using THP-1 monocytic cells and in vivo anti- inflammatory activity, was confirmed by carrageenan induced rat paw edema as model. Results: The activity guided isolation led to isolation of twelve compounds Which are: Stigmasterol [1] , β- sitosterol [2] , Lupeol [3] , Gallic acid [4] , Quercetin [5] , β- amyrin [6] , Oleanolic acid [7] , β- amyrin acetate [8] , 4- hydroxyl butyl cinnamate [9] , 4- hydroxyl cinnamic acid [10] , Caffeic acid, [11] and Kaempferol [12] respectively. Among the isolates, the compounds 4 and 5 displayed potent radical scavenging activity with an IC 50 values of 0.43 and 0.067 ΅g/ml. The compounds 4, 5 and 10 showed significant anti-diabetic activities. while lupeol [3] showed potent IL-1 β activity inhibition in THP-1 monocytic cells and also displayed significant (p<0.0025) in vivo anti-inflammatory activity. Conclusion: Inbrief, we isolated twelve compounds from both the species. Collectively, our results suggested that aromatic compounds showed good anti-oxidant and anti-diabetic activities. Abbreviations used: DPPH: 2,2- Diphenyl-1-1-picryl hydrazyl, ABTS: 2,2- Azinobis-3-ethylbenzo thiazoline-6-sulfonic acid, THP-1: Human leukaemia monocytic cell line, IL-1β: Interleukin-1β, IC 50 - Inhibitory concentration 50%.
  2,849 177 1
Potential therapeutic agents for the treatment of fatty degeneration of liver and atheromatous plaques: An experimental study in rats
Sibghatullah Muhammad Ali Sangi
July 2016, 12(47):414-423
DOI:10.4103/0973-1296.191444  PMID:27761068
Background: Since long high fat diet (HFD) is being blamed for causing fatty degeneration of liver and formation of atheromatous plaques. At present, no proper pharmacotherapy is available for both the conditions. In this study, different substances containing monounsaturated fatty acids were used to observe their protective effects in the HFD induced damage to liver and coronary vessels. Objectives: To discover effective therapeutic agents for HFD induced fatty degeneration of liver and atheromatous plaques. Materials and Methods: The study was conducted from September 2015 to April 2016. In this study, rats were divided into nine groups according to dietary regimen. Each group comprised six rats. Saturated fat was given in the form of butter, and unsaturated fat was given in the form of corn oil, olive oil, Nigella sativa oil, and crushed garlic. Serum samples were taken to estimate lipid profile, liver functions, cardiac functions, and kidney functions. Visceras were removed after animal sacrifice, and histopathological examination was done. Results and Conclusion: During the study period, the weight of animals changed significantly in some groups. Those animals which were given crushed garlic along with high saturated fat diet, showed protection against accumulation of lipids in the hepatocytes. Olive oil and Nigella sativa oil were comparatively less effective. Abbreviations used: HFD: High Fat Diet; NS: Nigella Sativa; TQ: Thymoquinone; KFMRC: King Fahad Medical Research Center; BUN: Blood Urea Nitrogen; BNF: Buffered Neutral Formalin; G: Group
  2,837 166 -
Evaluation of the genotoxicity and cytotoxicity of semipurified fractions from the Mediterranean brown algae, Dictyopteris membranacea
Najoua Akremi, Davie Cappoen, Roel Anthonissen, Abderrahman Bouraoui, Luc Verschaeve
July 2016, 12(47):395-399
DOI:10.4103/0973-1296.188318  PMID:27761065
Dictyopteris membranacea, a species of Mediterranean brown algae,is believed to have potential pharmacological and nutritional applications. However, such potentials only make sense when devoid of any adverse health consequences. The present study should be seen in this context. It aimed at evaluating the genotoxicity and cytoxicity of its organic extract (F0) and semi purified fractions (F 4, F 5, and F 6).Extracts were tested using the bacterial Vitotox® test and micronucleus assay in different concentrations (from 1.25 μg/mL up to 100 μg/mL, depending on the test and the extract). Applied concentrations were based on a preliminary dose-finding test with the neutral red uptake assay. The results show that all extracts were not genotoxic in the presence or absence of a rat metabolic enzyme fraction (S9). This is encouraging and justifies further investigations on the therapeutic and other values of this algae.
  2,665 196 -
Therapeutic effect of total saponins from Dioscorea nipponica makino on gouty arthritis based on the NF-κB signal pathway: An In vitro study
Qi Zhou, Shumin Liu, Donghua Yua, Ning Zhang
Jul-Sep 2016, 12(47):235-240
DOI:10.4103/0973-1296.186344  PMID:27601855
Objective: Dioscorea nipponica Makino is one of the most common used traditional Chinese drugs which are used to treat gouty arthritis (GA). Nuclear factor-κB (NF-κB) pathway plays an important role during this process. In the present study, we investigated the effects of total saponins from D. nipponica Makino (TDN) on NF-κB pathway in interleukin-1β (IL-1β) induced fibroblast-like synoviocytes (FLS). Materials and Methods: FLS were divided into three groups: Normal group, model group, which was given 10 μg/L IL-1β to induce the proliferation, and TDN group (10 μg/L IL-1β +100 μg/L TDN). 1 h, 24 h, 48 h, and 72 h after treating, immune fluorescence method was used to detect the cell location of NF-κB p65. Electrophoretic mobility shift assay was used to detect the activation of NF-κB p65. Western blot method was used to detect the protein expressions of NF-κB p65, IκBα, and p-IκBα. Results: TDN could inhibit the activation and transfer of NF-κB p65. As time went on, the expression of NF-κB p65 in the cytoplasm was decreased while it was increased in the nucleus. The expression of p-IκBα was increased, whereas the expression of IκBα was not changed. TDN could regulate these abnormal expressions. Conclusion: TDN may treat GA by regulating NF-κB signal pathway. SUMMARY
  • TDN could inhibit the transfer of NF-κB p65.
  • TDN could inhibit the activation of NF-κB p65.
  • TDN could inhibit the expression of p-IκBα
Abbreviations used: TDN: Total saponins from Dioscorea nipponica Makino, GA: Gouty arthritis, FLS: Fibroblast-like synoviocytes, IL-1β: Interleukin-1 beta, IF: Immune fluorescence, EMSA: Electrophoretic mobility shift assay, WB: Western blot.
  2,740 98 1
Human cytochrome P450 enzyme modulation by Gymnema sylvestre: A Predictive Safety evaluation by LC-MS/MS
Bera Rammohan, Karmakar Samit, Das Chinmoy, Saha Arup, Kundu Amit, Sarkar Ratul, Karmakar Sanmoy, Adhikari Dipan, Sen Tuhinadri
July 2016, 12(47):389-394
DOI:10.4103/0973-1296.191441  PMID:27761064
Background: Traditionally GS is used to treat diabetes mellitus. Drug-herb interaction of GS via cytochrome P450 enzyme system by substrate cocktail method using HLM has not been reported. Objective: To evaluate the in-vitro modulatory effects of GS extracts (aqueous, methanol, ethyl acetate, chloroform and n-hexane) and deacylgymnemic acid (DGA) on human CYP1A2, 2C8, 2C9, 2D6 and 3A4 activities in HLM. Material and Methods: Probe substrate-based LCMS/MS method was established for all CYPs. The metabolite formations were examined after incubation of probe substrates with HLM in the presence or absence of extracts and DGA. The inhibitory effects of GS extracts and DGA were characterized with kinetic parameters IC50 and Ki values. Results: GS extracts showed differential effect on CYP activities in the following order of inhibitory potency: ethyl acetate > Chloroform > methanol > n-hexane > aqueous > DGA. This differential effect was observed against CYP1A2, 2C9 and less on CYP3A4 and 2C8 but all CYPs were unaffected by aqueous extract and DGA. The ethyl acetate and chloroform extract exhibited moderate inhibition towards CYP1A2 and 3A4. The aqueous extract and DGA however showed negligible inhibition towards all five major human CYPs with very high IC50 values (>90μg/ml). Conclusion: The results of our study revealed that phytoconstituents contained in GS, particularly in ethyl acetate and chloroform extracts, were able to inhibit CYP1A2, 3A4 and 2C9. The presence of relatively small, lipophillic yet slightly polar compounds within the GS extracts may be attributed for inhibition activities. These suggest that the herb or its extracts should be examined for potential pharmacokinetic drug interactions in vivo. Abbreviations used: GS: Gymnema sylvestre, GSE: Gymnema sylvestre extract, DGA: deacyl gymnemic acid, CYP: cytochrome P450, DMSO: dimethylsulphoxide, HLM: human liver microsomes, LC-MS/MS: liquid chromatography tandem mass spectroscopy, NADPH: reduced nicotinamide adeninedinucleotide phosphate, NRS: nicotinamide adeninedinucleotide phosphate regenerating system, CHE: chloroform extract, EAE: ethyl acetate extract, NHE- n-hexane extract, AE: aqueous extract, ME: methanol extract,
  2,683 135 -
Rapid determination of puerarin by near-infrared spectroscopy during percolation and concentration process of puerariae lobatae radix
Xue Jintao, Yang Quanwei, Jing Yun, Liu Yufei, Li Chunyan, Yang Jing, Wu Yanfang, Li Peng, Wan Guangrui
Jul-Sep 2016, 12(47):188-192
DOI:10.4103/0973-1296.186350  PMID:27601848
Background: Gegen (Puerariae Labatae Radix) is one of the important medicines in Traditional Chinese Medicine. The studies showed that Gegen and its preparation had effective actions for atherosclerosis. Objective: Near-infrared (NIR) was used to develop a method for rapid determination of puerarin during percolation and concentration process of Gegen. Materials and Methods: About ten batches of samples were collected with high-performance liquid chromatography analysis values as reference, calibration models are generated by partial least-squares (PLS) regression as linear regression, and artificial neural networks (ANN) as nonlinear regression. Results: The root mean square error of prediction for the PLS and ANN model was 0.0396 and 0.0365 and correlation coefficients (r2) was 97.79% and 98.47%, respectively. Conclusions: The NIR model for the rapid analysis of puerarin can be used for on-line quality control in the percolation and concentration process. SUMMARY
  • Near-infrared was used to develop a method for on.line quality control in the percolation and concentration process of Gegen
  • Calibration models are generated by partial least.squares.(PLS) regression as linear regression and artificial neural networks.(ANN) as non.linear regression
  • The root mean square error of prediction for the PLS and ANN model was 0.0396 and 0.0365 and correlation coefficients.(r2) was 97.79% and 98.47%, respectively.
Abbreviations used: NIR: Near-Infrared Spectroscopy; Gegen: Puerariae Loabatae Radix; TCM: Traditional Chinese Medicine; PLS: Partial least-squares; ANN: Artificial neural networks; RMSEP: Root mean square error of validation; R2: Correlation coefficients; PAT: Process analytical technology; FDA: The Food and Drug Administration; Rcal: Calibration set; RMSECV: Root mean square errors of cross-validation; RPD: Residual predictive deviation; SLS: Straight Line Subtraction; MLP: Multi-Layer Perceptron; MSE: Mean square error. Wan Guangrui
  2,615 97 -
Postprandial antihyperglycemic and antioxidant activities of Acalypha indica Linn stem extract: An in-vivo study
Charles Lekhya Priya, KV Bhaskara Rao
July 2016, 12(47):475-481
DOI:10.4103/0973-1296.191461  PMID:27761078
Background: α-glucosidase inhibitors controls postprandial hyperglycemia (PPHG) by lowering sharp rise in blood glucose levels after ingestion of carbohydrate rich meal in type 2 diabetic (T2D) individuals. Acalypha indica commonly known as Indian copper leaf is used in traditional medicinal system to treat various diseases. In our previous in-vitro investigation, methanolic extract of A. indica stems (AIS) proved to be an effective a-glucosidase inhibitor, antioxidant, and well tolerated in acute and subchronic toxicity studies in albino wistar rats Objective: In this perspective, this study was designed to evaluate postprandial antihyperglycemic potential of AIS in maltose, sucrose, and glucose loaded streptozotocin (STZ)-induced normal and diabetic rats. As, the acute hyperglycemia at postprandial period has more triggering effect on oxidative stress, study was also aimed to evaluate the antioxidant potential of AIS on STZ-induced Albino-Wistar rats. Materials and Methods: Rats were treated with AIS (300-600 mg/kg b.w.) to investigate effect of AIS in controling PPHG after carbohydrate loading. Hepatoprotective activity of AIS is evaluated in diabetic rats by treating them at the dosages 300-600 mg/kg b.w. Results: Studies revealed 69.10 and 80.35% blood glucose-lowering effect of AIS in maltose and sucrose loaded diabetic rats in comparison with the diabetic control group. AIS recovered the liver damage caused by streptozotocin Conclusion: The present study confirmed high potential of AIS in controling PPHG by inhibiting a-glucosidase enzyme in maltose and sucrose loaded diabetic rats. AIS also exhibited hepatoprotective activity in STZ-induced diabetic rats. Thus, AIS could be used as a nutraceutical supplement to treat T2D effectively. Abbreviations used: AIS: Acalypha indica Stems, ALP: Alkaline Phosphatase, b/w: Body Weight, PPHG: Postprandial hyperglycemia, SE: Standard Error, SGOT: Serum glutamate oxaloacetate transaminase, SGPT: Serum glutamate pyruvate transaminase, SOD: Superoxide dismutase, STZ: Streptozotocin, TB: Total Bilirubin, T2D: Type 2 diabetes
  2,471 232 -
Elicitation of phenolics from the micropropagated endangered medicinal plant Calligonum polygonoides L. (Polygonoaceae)
Asmaa I Owis, Nada S Abdelwahab, Adel A Abul-Soad
July 2016, 12(47):465-470
DOI:10.4103/0973-1296.191458  PMID:27761076
Background: Calligonum polygonoides L. subsp. comosum (L'Hér.) Sosk. is a plant species belonging to family Polygonaceae. Susceptibility to threaten, presence of various chemical constituents, and many medicinal effects reported for this plant in addition to rareness of in vitro culture studies have fuelled the need for its micropropagation and phytochemical investigations of the produced cultures. Objectives: To employ in vitro culture technique for ex situ conservation of C. polygonoides, using the fruit as an explant; establish callus and cell suspension cultures from in vitro germinated plantlets; investigate the production of phenolics through callus, redifferentiated shoot, and cell suspension cultures; attempt to enhance cell capacity to accumulate phenolics using salicylic acid and yeast extract and provide a brief demonstration of biosynthetic pathway leading to phenolic production. Materials and Methods: Modified Murashige and Skoog media supplemented with growth hormones such as kinetin, 1-naphthaleneacetic acid, 6-benzylaminopurine, and indole-3-acetic acid were used to establish callus, redifferentiated shoots, and cell suspension cultures. Elicitation of cell suspension culture was performed using salicylic acid and yeast extracts. A reversed phase-high performance liquid chromatography method for determination of phenolic content in the aforementioned cultures was developed. Results: The unorganized callus and cell suspension cultures contained fewer amounts of phenolic compounds than redifferentiated shoots. Elicitation produced massive quantitative reprogramming of phenolic content. Conclusion: The present study offers an alternative and renewable source for this valuable natural plant, provide a chance to improve secondary metabolite yield and serve as a useful tool for studying the biosynthesis of these compounds and its regulation in plant cells. Abbreviation used: H 2 O 2: Hydrogen peroxide, Kin: Kinetin, NAA: Naphthaleneacetic acid, BAP: 6-benzylaminopurine, IAA: Indole-3-acetic acid, HPLC: High-performance liquid chromatography.
  2,456 206 -
Catha edulis (khat) induces apoptosis in Madin-Darby bovine kidney cell line
Hussein M Ageely, Ahmed E Agag, Syam Mohan, Atef Shehata
July 2016, 12(47):454-459
DOI:10.4103/0973-1296.191456  PMID:27761074
Background: Khat (Catha edulis) is a controversial plant having a euphoretic effect, at the same time part of culture in many countries such as Africa and Arabian Peninsula. The presence of amphetamine-like substance, cathinone and cathine make this plant banned in many countries. Many neurological and other system related studies have been carried out in this plant, but the lack of toxicity studies are there especially the mechanism. Objective: In this study, Madin-Darby Bovine Kidney cell line was used as an in vitro model to study the cell death mechanism. Crude extract of fresh Khat plant leaves were prepared and exposed to cells. Materials and Methods: Trypan blue assay, phase-contrast microscopy, fluorescent microscopy, clonogenic assay, annexin-V assay, and hematoxylin and eosin (H and E) staining were employed to check the objectives. Results: Reductions in cellular viability were observed at concentrations above 1.25 mg/ml while using Trypan blue assay. The results of the clonogenic assay had shown that the untreated control with the highest number of colonies (100% survival) and the 0.1562 concentration could not prevent the colony formation significantly. The high concentrations reduced the colony formation at concentration dependent manner 27.4% and 24.9%, for 0.625 mg/ml and 1.25 mg/ml concentrations, respectively. The acridine orange/ethidium bromide experiment had observed the cells were intact with round nucleus while the apoptosis features such as blebbing and nuclear chromatin condensation were clearly observed in treatment. The shrinkage of cells was clearly observed in H and E staining. Conclusion: In addition, annexin-V binding confirmed the presence of apoptosis significantly on Khat treatment. Abbreviation used: PS: Phosphatidylserine (PS); MDBK: Madin-Darby Bovine Kidney; DMEM: Dulbecco's modified Eagle's medium; PI: propidium iodide; EB: ethidium bromide; PBS: Phosphate Buffer saline; FITC: fluorescein isothiocyante; TUNEL: Terminal deoxynucleotidyl transferase dUTP nick end labeling.
  2,455 176 -
Responsive surface methodology optimizes extraction conditions of industrial by-products, Camellia japonica seed cake
Jae Kyeom Kim, Ho-Jeong Lim, Mi-So Kim, Soo Jung Choi, Mi-Jeong Kim, Cho Rong Kim, Dong-Hoon Shin, Eui-Cheol Shin
Jul-Sep 2016, 12(47):184-187
DOI:10.4103/0973-1296.186339  PMID:27601847
Background: The central nervous system is easily damaged by oxidative stress due to high oxygen consumption and poor defensive capacity. Hence, multiple studies have demonstrated that inhibiting oxidative stress-induced damage, through an antioxidant-rich diet, might be a reasonable approach to prevent neurodegenerative disease. Objective: In the present study, response surface methodology was utilized to optimize the extraction for neuro-protective constituents of Camellia japonica byproducts. Materials and Methods: Rat pheochromocytoma cells were used to evaluate protective potential of Camellia japonica byproducts. Results: Optimum conditions were 33.84 min, 75.24%, and 75.82°C for time, ethanol concentration and temperature. Further, we demonstrated that major organic acid contents were significantly impacted by the extraction conditions, which may explain varying magnitude of protective potential between fractions. Conclusions: Given the paucity of information in regards to defatted C. japonica seed cake and their health promoting potential, our results herein provide interesting preliminary data for utilization of this byproduct from oil processing in both academic and industrial applications. SUMMARY
  • Neuro-protective potential of C. japonica seed cake on cell viability was affected by extraction conditions
  • Extraction conditions effectively influenced on active constituents of C. japonica seed cake
  • Biological activity of C. japonica seed cake was optimized by the responsive surface methodology.
Abbreviations used: GC-MS: Gas chromatography-mass spectrometer, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, PC12 cells: Pheochromocytoma, RSM: Response surface methodology. Eui-Cheol Shin
  2,505 87 1
Phytochemicals and cytotoxicity of Launaea procumbens on human cancer cell lines
Preeti Rawat, Lokesh M Saroj, Anil Kumar, Tryambak D Singh, SK Tewari, Mahesh Pal
July 2016, 12(47):431-435
DOI:10.4103/0973-1296.191452  PMID:27761070
Background: The plant Launaea procumbens belongs to the family Asteraceae and traditionally used in the treatment rheumatism, kidney, liver dysfunctions and eye diseases. In the present study Phytochemical analysis and fractions of methanolic extract of L. procumbens leaves were tested in vitro for their cytotoxicity. Objectives: Phytochemical analysis and cytotoxic activity of methanolic extract and fractions of Launaea procumbens against four cancer cell lines K562, HeLa, MIA-Pa-Ca-2 and MCF-2 by SRB assay. Materials and Methods: Powdered leaves of Launaea procumbens were extracted sequentially with hexane, ethyl acetate, butanol and water by cold extraction. Phytochemical analysis and cytotoxicity assay were carried out for these fractions using SRB assay against four human cancer cell lines, namely leukemia (K562), cervix (HeLa), pancreatic (MIA-Pa-Ca-2) and breast (MCF-7). Results: Ethyl acetate extract exerts potent cytotoxicity against human leukemia (K562), cervix (HeLa) and breast (MCF-7) cell lines IC 50 value of 25.30±0.50, 19.80±0.10 and 36.90±4.90 μg/ ml respectively. Moderately cytotoxic effect found in hexane extract IC 50 value of 41±8 and 48.20±0.50 μg/ ml against leukemia (K562), and breast (MCF-7) cancer cell line respectively. The Chemical composition analyzed by GC-MS showed considerable differences in solvent fractions of Launaea procumbens. Conclusion: This study revealed the cytotoxic potential of ethyl acetate and hexane fractions of L.procumbens leaves on different cancer cell lines. Abbreviations used: SRB: Sulforhodamine B assay, MW: Molecular weight
  2,291 297 -
Influence of extracting solvent on pharmacological activity and cytotoxicity of Polygonum minus, a commonly consumed herb in Southeast Asia
Parayil Varghese Christapher, Subramani Parasuraman, Palanimuthu Vasanth Raj, Sultan Ayesh Mohammed Saghir, Mohd. Zaini Asmawi, Murugaiyah Vikneswaran
July 2016, 12(47):424-430
DOI:10.4103/0973-1296.191451  PMID:27761069
Objective: To investigate the antihyperlipidemic, antioxidant, and cytotoxic effect of aqueous and methanol extract of leaves of Polygonum minus. Materials and Methods: Acute antihyperlipidemic effect was studied on chemically induced hyperlipidemic rat model. Treated groups received aqueous and methanol extract of leaves of P. minus respectively (1000 mg/kg; oral) whereas standard treated group received atorvastatin (60 mg/kg; oral) for 3 consecutive days. Blood samples were collected at fixed intervals for lipid profile analysis. Antioxidant effects were studied using 1,1-diphenyl-2-picrylhydrazyl, 2,2-azinobis 3-ethylbenzothiazoline 6-sulfonate, and ferric reducing antioxidant power assays. The total flavonoids content and total phenolic contents were also estimated. Cytotoxicity of both extracts was studied on one normal and three cancer cell lines using 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay method. Results: The methanol extract showed significant reduction in total cholesterol (P < 0.001), triglycerides (P < 0.01), LDL (P < 0.05), VLDL (P < 0.01), atherogenic index (P < 0.001), and elevation of HDL (P < 0.05) levels than the aqueous extract. Similarly, the antioxidant investigations also demonstrated that the methanol extract had higher antioxidant capacity than aqueous extract. Both extracts were not toxic to normal (EA.hy926) as well as to cancer (HCT116, HT29, and HeLa) cells. Significant correlation was demonstrated between total phenolic and total flavonoids contents with the antioxidant activity but not with the antihyperlipidemic effect, suggesting other groups of chemical constituents may be mainly responsible for the antihyperlipidemic effect of this plant. Conclusion: The study demonstrated that the presence and extent of bioactivities are influenced by solvents used for extraction. This study confirmed the antihyperlipidemic effect of leaves of P. minus in acute hyperlipidemic rat model. Abbreviations used: CVDs: Cardiovascular diseases, LDL: Low-density lipoprotein, DDPH: 1 ,1-Diphenyl-2-picrylhydrazyl radical, TPTZ : 2,4,6,-tris(1-pyridyl)-5-triazine, ABTS : 2,2?-Azino-di-[3-ethylbenzthiazoline Sulfonate], HDL: High-density lipoprotein, VLDL: Very low-density lipoprotein, TC: Total cholesterol, TG: Triglycerides, EC 50: Half maximal effective concentration, LD 50: Median lethal dose.
  2,242 181 1
Antihepatotoxic effect and metabolite profiling of Panicum turgidum extract via UPLC-qTOF-MS
Mohamed A Farag, Ahlam M El Fishawy, Sayed A El-Toumy, Khadiga F Amer, Ahmed M Mansour, Hala E Taha
July 2016, 12(47):446-453
DOI:10.4103/0973-1296.191455  PMID:27761073
Background: Panicum turgidum, desert grass, has not reported any detailed phytochemical or biological study as yet Objective: To establish P. turgidum secondary metabolite profile and to assess its antihepatotoxic effect Materials and Methods: Ultra-performance liquid chromatography (UPLC) coupled to quadrupole high-resolution time of flight mass spectrometry (qTOF-MS) was used for large-scale secondary metabolites profiling in P. turgidum extract, alongside assessing median lethal dose (LD50) and hepatoprotective effect against carbon tetrachloride (CCl 4 ) intoxication Results: A total of 39 metabolites were identified with flavonoids as the major class present as O/C-glycosides of luteolin, apigenin, isorhamnetin and naringenin, most of which are first time to be reported in Panicum sp. Antihepatotoxic effect of P. turgidum crude extract was revealed via improving several biochemical marker levels and mitigation against oxidative stress in the serum and liver tissues, compared with CCl4 intoxicated group and further confirmed by histopathological examination. Conclusion: This study reveals that P. turgidum, enriched in C-flavonoids, presents a novel source of safe antihepatotoxic agents and further demonstrates the efficacy of UPLC-MS metabolomics in the field of natural products drug discovery. Abbreviations used: UPLC: Ultra-performance liquid chromatography (UPLC), LD50: median lethal dose, MDA: malondialdehyde, GSH: glutathione reductase, CAT: catalase, SOD: superoxide dismutase, ALT: alanine aminotransferase, AST: aspartate aminotransferase, ALP: alkaline phosphatase, TG: triglycerides.
  2,251 133 1
Genetic and chemical profiling of Gymnema sylvestre accessions from central India: Its implication for quality control and therapeutic potential of plant
Ashutosh Kumar Verma, Sunita Singh Dhawan, Seema Singh, Kumar Avinash Bharati, Jyotsana
July 2016, 12(47):407-413
DOI:10.4103/0973-1296.191443  PMID:27761067
Background: Gymnema sylvestre, a vulnerable plant species, is mentioned in Indian Pharmacopeia as an antidiabetic drug Objective: Study of genetic and chemical diversity and its implications in accessions of G. sylvestre Materials and Methods: Fourteen accessions of G. sylvestre collected from Central India and assessment of their genetic and chemical diversity were carried out using ISSR (inter simple sequence repeat) and HPLC (high performance liquid chromatography) fingerprinting methods Results: Among the screened 40 ISSR primers, 15 were found polymorphic and collectively produced nine unique accession-specific bands. The maximum and minimum numbers of amplicones were noted for ISSR-15 and ISSR-11, respectively. The ISSR -11 and ISSR-13 revealed 100% polymorphism. HPLC chromatograms showed that accessions possess the secondary metabolites of mid-polarity with considerable variability. Unknown peaks with retention time 2.63, 3.41, 23.83, 24.50, and 44.67 were found universal type. Comparative hierarchical clustering analysis based on foresaid fingerprints indicates that both techniques have equal potential to discriminate accessions according to percentage gymnemic acid in their leaf tissue. Second approach was noted more efficiently for separation of accessions according to their agro-climatic/collection site Conclusion: Highly polymorphic ISSRs could be utilized as molecular probes for further selection of high gymnemic acid yielding accessions. Observed accession specific bands may be used as a descriptor for plant accessions protection and converted into sequence tagged sites markers. Identified five universal type peaks could be helpful in identification of G. sylvestre-based various herbal preparations. Abbreviations used: HPLC: High Performance Liquid Chromatography, ISSR: Inter Simple Sequence Repeats, CTAB: Cetyl Trimethylammonium Bromide, DNTP: Deoxynucleotide Triphosphates
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Expression of cytochrome P450s in the liver of rats administered with Socheongryong-tang, a traditional herbal formula
Seong Eun Jin, Hyekyung Ha, Chang-Seob Seo, Hyeun-Kyoo Shin, Soo-Jin Jeong
Jul-Sep 2016, 12(47):211-218
DOI:10.4103/0973-1296.186340  PMID:27601852
Objective: The purpose of this study was to investigate the potential influences of Socheongryong-tang(SCRT) on the messenger ribonucleic acid (mRNA) and protein expression of cytochrome P450 (CYP450) in vivo. Materials and Methods: SCRT was orally administered to either male or female Sprague-Dawley rats once daily at doses of 0, 1000, 2000, or 5000 mg/kg/day for 13 weeks. The mRNA expression of CYP450s (CYP1A1, 1A2, 2B1/2, 2C11, 2E1, 3A1, 3A2, and 4A1) in liver tissues was measured by reverse transcription polymerase chain reaction. And then, the protein expression of CYP1A1 and CYP2B1/2 in liver tissues was analyzed by the Western blot. Results: We found no significant influence in the mRNA expression of hepatic CYP1A2, 2C11, 2E1, 3A1, 3A2, and 4A1 after repeated administration of SCRT for 13 weeks. By contrast, the mRNA and protein expression of hepatic CYP1A1 was increased by repeated SCRT treatment in male rats, but not in female rats. The mRNA and protein expression of hepatic CYP2B1/2 in both genders was increased by administration of SCRT. Conclusion: A caution is needed when SCRT is co-administered with substrates of CYP2B1/2 for clinical usage. In case of male, an attention is also required when SCRT and drugs metabolized by CYP1A1 are taken together. Our findings provide information regarding the safety and effectiveness of SCRT when combined with conventional drugs. SUMMARY
  • Oral administration of Socheongryong-tang for 13 weeks did not affect the mRNA expression of hepatic CYP1A2, 2C11, 2E1, 3A1, 3A2, and 4A1
  • In male rats, oral administration of Socheongryong-tang for 13 weeks induced the mRNA and protein expression of hepatic CYP1A1 and CYP2B1/2
  • In female rats, oral administration of Socheongryong-tang for 13 weeks induced the mRNA and protein expression of hepatic CYP2B1/2.
Abbreviations used: SCRT: Socheongryong-tang, CYP450: Cytochrome P450, HPLC: High performance liquid chromatography, RT-PCR: Reverse transcription polymerase chain reaction. Soo-Jin Jeong
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