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October-December 2018
Volume 10 | Issue 4
Page Nos. 333-441

Online since Friday, October 26, 2018

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ORIGINAL ARTICLES  

Antithrombotic activity and saponin composition of the roots of Panax bipinnatifidus Seem. growing in Vietnam Highly accessed article p. 333
Vu Thi Thom, Nguyen Huu Tung, Dang Van Diep, Dang Thi Thuy, Nguyen Thi Hue, Dinh Doan Long, Bui Thanh Tung, Pham Thanh Huyen, Duong Thi Ly Huong
DOI:10.4103/pr.pr_58_18  
Background: Panax bipinnatifidus (PB) Seem. is a highly valuable and indigenous medicinal plant wildly distributed in the northwest region of Vietnam. The PB root has been used as a tonic in Vietnamese traditional medicine, but its chemical profile and pharmacological activity have not been extensively studied. Objective: In this study, we investigated the major saponin components and antithrombosis activity of the root of PB. Materials and Methods: Individual saponins were isolated by column chromatography. The structural elucidation of the isolated saponins was based on analyses of nuclear magnetic resonance and mass spectrometry spectra. Chromatographic fingerprint analysis of the isolated compounds was run on an Agilent 1260 high-performance liquid chromatography (HPLC) system. To evaluate antithrombotic activity, in vitro anticoagulant, platelet aggregation, and clot lysis effects were successively tested by the respective protocols. Results: Two principal saponins were isolated from the butanolic fraction of the root of PB, and their structures were identified as stipuleanoside R2 (1) and araloside A methyl ester (2). The HPLC analysis suggested that the two isolated saponins are major saponins of the title plant. The biological testing showed that both the crude and butanolic extracts of PB exhibited significantly antiplatelet aggregation activity. The efficacy linearly increased according to the tested doses (0.5–5 mg/mL). On anticoagulant activity, only butanolic extract exhibited positive action via prolonged activated partial thromboplastin time and prothrombin time at the high dose. Conclusion: These results suggested that PB Seem. might bring benefits for preventing cardiovascular events through inhibition of platelet aggregation. Abbreviations Used: ADP: Adenosine diphosphate; APTT: Activated partial thromboplastin time; DMSO: Dimethyl sulfoxide; ESI-MS: Electrospray ionization-mass spectrometry; HPLC: High-performance liquid chromatography; LC-MS/MS: Liquid chromatography-tandem mass spectrometry; MAPK: Mitogen-activated protein kinase; MPA: Maximum platelet aggregation; NIMM: National Institute of Medicinal Materials; NMR: Nuclear magnetic resonance; PPP: Platelet-poor plasma;; PRP: Platelet-rich plasma; PT: Prothrombin time; TLC: Thin-layer chromatography; VNU: Vietnam National University.
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Central nervous system activity of phenol-rich fraction of Piper sylvaticum roots p. 339
Akash Ved, Amresh Gupta, Om Prakash, Ajay Kumar Singh Rawat
DOI:10.4103/pr.pr_125_17  
Obective: Piper sylvaticum Roxb. is an important folk medicine in Indian Traditional System of Medicine widely used by different tribes in many countries. In the present study, the anticonvulsant activity of extract/fractions of Piper sylvaticum (PS) roots was investigated. Materials and Methods: 70% ethanolic extract of PS roots was successively extracted using hexane and ethyl acetate to prepare various fractions. Total phenol content was found at maximum 324.65 mg/gallic acid equivalent/g in ethyl acetate fraction (EAF) (phenol-rich fraction [PRF]). High-performance thin-layer chromatography fingerprinting profiling of PS roots was performed. The anticonvulsant properties of the EAF of roots of PS were examined by maximal electroshock method as compared to standard phenytoin (25 mg/kg body weight). Result: It was found that EAF shows potent anticonvulsant activity at different dose levels against maximum electroshock seizure-induced convulsions in Swiss albino mice. Conclusion: From the observation, it can be concluded that the current study has expressed that the phenol-rich EAF of the ethanolic extract of the roots of PS has shown the dose-dependent anticonvulsant effect in mice. The anticonvulsant potential may be due to the presence of phenolic compounds in PRF. The outcomes suggested a high potential for application of EAF of PS root as an anticonvulsant agent. Abbreviations Used: HPTLC: High-performance thin-layer chromatography, OECD: Organization for Economic Co-operation and Development, GAE: Gallic acid equivalent, PRF: Phenol-rich fraction.
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The effect of capsicum oleoresin on nitric oxide production and nitric oxide synthase gene expression in macrophage cell line p. 343
Srinivasan Manthra Prathoshni, Roy Anitha, Thangavelu Lakshmi
DOI:10.4103/pr.pr_46_18  
Background: Nitric oxide (NO) is an inflammatory agent produced by macrophages. It also acts as a neurotransmitter. However, overproduction of NO results in acute or chronic inflammation. Capsicum is well known for its anti-oxidant, anti–inflammatory, and anticancer properties. Objective: The objective of this study was to evaluate the effect of capsicum oleoresin on NO production and NO synthase gene expression in lipopolysaccharide (LPS) stimulated RAW 264.7 macrophage cell line. Materials and Methods: Macrophage RAW 264.7 cells were obtained from the National Centre for Cell Science, Pune with Passage no 16. RAW macrophages were treated with 12.5 μg/ml, 25 μg/ml, and 50 μg/ml of Capsicum extract with 1 μg/ml of LPS and incubated for 24 h. Results: When capsicum was added at three different doses of 12.5 μg/ml, 25 μg/ml, and 50 μg/ml, the inducible NO synthase (iNOS) levels was significantly suppressed, compared to that of LPS treatment only. The level of NO increased by LPS induction was significantly decreased in a dose-dependent manner when treated with different concentrations of capsicum extract and capsicum had a suppressing effect on iNOS gene expression in LPS – stimulated RAW 264.7 macrophage. Conclusion: This study concludes that capsicum oleoresin is good enough to suppress iNOS gene expression and NO production. Hence, it may be used in inflammatory conditions with excessive NO production. Abbreviations Used: NO: Nitric oxide, LPS: Lipopolysaccharide, iNOS: Inducible nitric oxide synthase, DMEM: Dulbecco's modified Eagle medium, MTT: 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide for, DMSO: Dimethyl sulfoxide, PBS: Phosphate buffer saline, FBS: Fetal bovine serum.
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Study of glucose uptake enhancing potential of fenugreek (Trigonella foenum graecum) leaves extract on 3T3 L1 cells line and evaluation of its antioxidant potential p. 347
Debarupa Hajra, Santanu Paul
DOI:10.4103/pr.pr_50_18  
Background: The incidence of diabetes mellitus (DM) is increasing at an alarming rate globally. There is a need for suitable natural alternatives for its treatment, as the regular use of drugs causes several secondary health issues. Aim: This study aimed to quantitatively evaluate the phenol and flavonoid content; antioxidant and anti-diabetic activity of fenugreek seeds and leaves in water and ethanolic extracts. Materials and Methods: Dried and powdered seeds and leaves of fenugreek were treated at different time and temperature combinations (100°C for 10 min; 80°C for 30 min; 60°C for 1 h; 37°C for 6 h; 24°C [room temperature] for 24 h; 100°C for 10 min followed by overnight soaking). Total phenolic content, total flavonoid content, 2,2-Diphenyl-1-picrylhydrazyl (DPPH), and glucose uptake assays on 3T3-L1 cell lines were performed on the extracts. Results: The results show that fenugreek leaves treated at 100°C for 10 min and overnight soaking yield extracts with the highest concentrations of desired phenols (46.08 ± 0.15 mg GAE/g sample) and flavonoid content (13.02 ± 0.44 mg/g sample) and antioxidant activity (DPPH) (45.41 ± 2.1%) with enhanced glucose uptake activity in 3T3-L1 cell lines. Conclusion: The phenolic and flavonoid content, DPPH radical scavenging activity and antidiabetic activity was highest in the water extract of fenugreek leaves treated at 100°C for 10 min and subsequent soaking for 24 h before filtration as compared to the other protocols tested. Furthermore, water extracts showed enhanced activity as compared to the ethanol extracts in case of both seeds and leaves and in all the treatment combinations. Abbreviations Used: 2NBDG: (2-(N-(7-Nitrobenz-2-oxa-1, 3-diazol-4-yl) Amino)-2-Deoxyglucose), DM: Diabetes mellitus, DMEM: Dulbecco's Modified Eagle's Medium, DPPH: 2,2-Diphenyl-1-picrylhydrazyl, GA: Gallic Acid, GAE: Gallic Acid Equivalents, PBS: Phosphate Buffered Saline, TFC: Total Flavonoid Content, TPC: Total Phenolic Content
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Zingiber zerumbet rhizomes extract exhibits faster open wound healing in rats p. 354
Asmah Hamid, Chong Pek Lian, Nur Hafiza Salleh Hudin, Ahmad Rohi Ghazali, Nurul Farhana Jufri
DOI:10.4103/pr.pr_14_18  
Background: Zingiber zerumbet rhizome ethyl acetate extract (ZZRE) has the potential to be developed as a wound-healing agent with its profound anti-inflammatory property. Objective: This study was conducted to evaluate the wound healing effects of 5% ZZRE on open wound in rats. Materials and Methods: Wistar rats (n = 90) were divided into three groups, negative control (treated with PBS), positive control (treated with solcoseryl gel), and 5% ethyl acetate extract of Z. zerumbet, respectively. Wounds were induced on the rat's dorsal surface on day 0. Measurements of wound closure, total protein, uronic acid, hexosamine, cathepsin B, hydroxyproline, and histological observations were conducted on the wound tissues. Results: ZZRE-treated group showed increment in wound closure, protein, and hydroxyproline level. The uronic acid and hexosamine levels of ZZRE-treated wounds significantly increased on the 3rd day (P < 0.05). Histological observations demonstrated formation of granulation tissue was faster and denser in ZZRE-treated group. Conclusion: ZZRE enhances open wound healing in rats and has potential to be developed as a healing agent in the future. Abbreviations Used: ZZRE: Zingiber zerumbet rhizomes extract, PDGF: Platelet-derived growth factor, TGF-β: Transforming growth factor-beta, PBS: Phosphate-buffered saline, EDTA: Ethylenediaminetetraacetic acid
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In vitro antioxidant and antidiabetic activity of oligopeptides derived from different mulberry (Morus alba L.) cultivars p. 361
Suchisree Jha, Saran Kumar Gupta, Phalguni Bhattacharyya, Amitava Ghosh, Palash Mandal
DOI:10.4103/pr.pr_70_18  
Background: Several plant peptides have been known to exhibit various pharmacological activities. However, the bioactivity of mulberry peptides is not yet reported. Objective: In the present study, the peptides from different mulberry cultivars, namely S1, V1, S1635, and Dudhiya, were isolated and assessed for their in vitro free radical scavenging capacity and inhibitory activity of two key enzymes (α-amylase [AA] and α-glucosidase [AG]) associated with type-II diabetes. Materials and Methods: The 0.5–3 kDa ranged oligopeptides were isolated using cation exchange chromatography and ultrafiltration system and further characterized by high-performance liquid chromatography and protein sequencer. The peptides were investigated for their in vitro antioxidant potential by 1,1-diphenyl-2 picrylhydrazyl, 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, and nitric oxide scavenging capacity and reducing power, metal chelating, and anti-lipid peroxidation activity along with in vitro antidiabetic activity by AA and AG inhibition. Results: The results revealed that the peptides were found to possess significant free radical scavenging as well as AA and AG inhibitory activity in a dose-dependent manner. Overall, the peptide isolated from S1 cultivar exhibited the most promising therapeutic potential. Further, the variation in amino acid composition of the oligopeptides could be associated with the observed variation in their bioactivity. Conclusion: These natural peptides may constitute an important part of the antioxidant defense system as well as antidiabetic agent and further could be used for the formulation of functional food and nutraceuticals. Abbreviation Used: HPLC: High-performance liquid chromatography, PMSF: Phenyl methyl sulfonyl fluoride; IAA: Indoleacetic acid, ABA: Abscisic acid, GA3: Gibberellic acid, DPPH: 1,1-diphenyl-2 picrylhydrazyl, ABTS: 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, AG: α-glucosidase, AA: α-amylase, DNSA: 3,5-Dinitrosalicylic acid, AAE: Ascorbic acid equivalent, TBARS: Thiobarbituric acid-reactive species.
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HPLC-DAD-ESI-MS/MS characterization of bioactive secondary metabolites from Strelitzia nicolai leaf extracts and their antioxidant and anticancer activities In vitro p. 368
Mosad Ghareeb, Amal Saad, Wafaa Ahmed, Laila Refahy, Sami Nasr
DOI:10.4103/pr.pr_89_18  
Background: Strelitzia nicolai Regel and Körn (Strelitziaceae) is native to Southern Africa whose phytochemistry and pharmacology were slightly investigated. Materials and Methods: In the current work, different solvent extracts of S. nicolai were screened for their chemical profiles through high-performance liquid chromatography coupled with diode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESI-MS/MS) analyses. Furthermore, their in vitro antioxidant, cytotoxic, and anticancer activities were evaluated using 2,2'-diphenyl-1-picrylhydrazyl radical (DPPH), 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) & ferric reducing antioxidant power (FRAP) and crystal violet staining (CVS) colorimetric assays, respectively. Results: HPLC-DAD-ESI-MS/MS analyses led to the identification of nineteen and eleven phenolic compounds from the ethyl acetate and n-butanol extracts, respectively including flavonoids (e.g., quercetin 3-(2 G-rhamnosylrutinoside, quercetin, quercetin-3-O-glucoside, kaempferol-3,7-O-dirhamnoside, isorhamnetin-3-O-rutinoside and kaempferol-3-O-glucoside), phenolic acids derivatives (e.g., chlorogenic acid glycoside, protocatechuic acid-O-glucoside and caftaric acid), chalcones (e.g., xanthoangelol), and phenylethanoids (e.g., ligstroside glucoside). Moreover, in the DPPH assay the IC50value of the most active ethyl acetate extract was 20.49 μg/mL, relative to 2.92 μg/mL of ascorbic acid. ABTS and FRAP results reinforced the results of DPPH assay. According to the National Cancer Institute criteria, the tested extracts showed weak to moderate cytotoxic activities with IC50values ranged from 65.23 to 451.29 μg/mL. Furthermore, the EtOAc and n-BuOH extracts showed a noticeable anticancer activity with CVS spectroscopic readings for liver hepatocellular carcinoma growth 0.806 and 0.684 at a concentration (125 μg/mL), as well as 0.730 and 0.618 at concentration (500 μg/mL), respectively against control at 1.022. Conclusion: The obtained results reveal the high efficacy of the phenolic-rich extracts from S. nicolai as naturally occurring antioxidant and anti-tumor agents. Abbreviations Used: HPLC-DAD-ESI-MS/MS: High-performance liquid chromatography-diode array detection-electrospray ionization-mass/mass; DPPH: 2,2'-Diphenyl-1-picrylhydrazyl radical; ABTS: 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulphonic acid); FRAP: Ferric reducing antioxidant power; TPTZ: Tripyridyl-s-triazine; FE: Ferrous equivalents; DMEM: Dulbecco's modified eagle's medium; DMSO: Dimethyl sulfoxide; EDTA: Ethylenediaminetetraacetic acid; PBS: Phosphate buffered saline; HepG-2: Liver hepatocellular carcinoma; CVS: Crystal violet stain; NCI: National cancer institute; Glu: Glucose; Rha: Rhamnose.
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Solubility of calcium oxalate and calcium phosphate crystallization in the presence of crude extract and fractions from Kelussia odoratissima Mozaff p. 379
Akram Torki, Tahereh Hosseinabadi, Sheida Fasihzadeh, Arezo Sadeghimanesh, Joko Priyanto Wibowo, Zahra Lorigooini
DOI:10.4103/pr.pr_68_18  
Background and Aim: Urinary tract stones are the third common urinary tract disease that may lead to renal failure. Kelussia odoratissima is traditionally used in the treatment of kidney stones in Chaharmahal va Bakhtiari region. This study was designed to investigate in vitro effect of crude extract and fractions of K. odoratissima on kidney stones (calcium oxalate and calcium phosphate). Materials and Methods: A total of 70% ethanolic extract of K. odoratissima was prepared by maceration method followed by liquid–liquid extraction with hexane, chloroform, ethyl acetate, and saturated n-butanol to get four fractions. Calcium oxalate and calcium phosphate were synthesized and then were treated with hydroalcoholic extract and fractions. Their effects on the dissolution of generated stone were assayed by calcium kit. The stones were synthesized and confirmed by fourier-transform infrared technique. Results: Results showed that total extract and its fractions had significant potency to dissolve calcium oxalate and calcium phosphate crystals. The results indicate the higher potency of fractions containing nonpolar compounds to dissolve calcium phosphate and calcium oxalate stones compared to the fractions containing polar compounds. n-butanolic fraction had the least effect and hexane fraction had the greatest effect on the calcium phosphate stones. Furthermore, the total extract has less dissolution ability, compared to the fractions. Conclusion: The obtained results of this study exhibited that the use of K. odoratissima extract and its fractions could help to dissolve urinary stones. Therefore, it can be effective in prevention and treatment of kidney stones on people who are prone to the formation of calcium oxalate and phosphate stones. Abbreviations Used: Caox: Calcium Oxalate, Caph: Calcium Phosphate, K. odoratissima: Kelussia odoratissima Mozaff
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Antioxidant and α-glucosidase inhibitory activities and gas chromatography–mass spectrometry profile of salak (Salacca zalacca) fruit peel extracts p. 385
Mohammed S. M. Saleh, Mohammad Jamshed Siddiqui, Siti Z Mat So'ad, Suganya Murugesu, Alfi Khatib, Mokhlesur M Rahman
DOI:10.4103/pr.pr_7_18  
Background: Salacca zalacca or better known as salak fruit is widely distributed in tropical and subtropical countries, and it is traditionally used to treat diabetes. This study was aimed to investigate the salak peel extracts for their biological and chemical activities. Also, the chemical profile of the most promising extract was analysed on gas chromatography- mass spectrometry (GC-MS). Materials and Methods: The peel extracts were prepared by maceration process at room temperature with different ratio of ethanol/water. All the extracts were determined for their α-glucosidase inhibitory activity using α-glucosidase enzyme. The antioxidant activities of the extracts were determined through their Ferric reducing antioxidant power assay (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH). The chemical constituents of salak peel extracts were analysed using gas chromatography–mass spectrometry (GC MS). Results: Phytochemical screening showed the presence of phenolic and flavonoid contents in all the extracts. About 100% ethanol extract shows the highest phenolic content (116.70 ± 0.764 μg/mL) while 60% ethanol extract has the lowest content 18.65 ± 1.155 μg/ml using gallic acid as a reference. 100% ethanol extract was observed to exhibit highest radical scavenging, ferric reducing antioxidant power (FRAP), and α-glucosidase inhibitory activities (IC50: 49.45 ± 3.87 μg/mL, 144.81 ± 3.72 μg AAE/g, IC50: 11.62 ± 0.67b μg/mL), respectively. Water extracts had the lowest FRAP, radical scavenging activity as well as α-glucosidase activity. The phytochemical investigation on GC-MS showed the presence of active compounds in salak fruit peel extracts. Conclusion: Salak fruit peels showed the highest antioxidant as well as α-glucosidase inhibitory activities. Phytochemical analysis on GC-MS confirms the presence of gallic acid, linoelaidic acid, palmitic acid, α-tocopherol, and steric acid which may contribute to α-glucosidase inhibitory activity. Abbreviations Used: GC-MS: Gas Chromatography–Mass Spectroscopy; FRAP: Ferric reducing antioxidant power assay; TPTZ: 2,4,6-tris (2-pyridyl)-s-triazine; DPPH: 2,2-diphenyl-1-picrylhdroxyl; MSTFA: N-methyl-N-(trimethylsilyl)-trifluoroacetamide; TPC: Total phenolic content; DMSO: Dimethyl sulfoxide; ANOVA: Analysis of variance.
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Protective effect of Ferulago angulata (Schltdl.) Boiss. hydroalcoholic extract against acetic acid-induced colitis in rat: Role of pro-inflammatory cytokines p. 391
Fatemeh Farzaei, Fatemeh Heydarpour, Mohammad Hosein Farzaei, Zahra Minush Siavash Haghighi, Zahra Abbasabadi, Ehsan Sadeghi
DOI:10.4103/pr.pr_9_18  
Background: Ulcerative colitis (UC) is a chronic recurrent inflammatory disease with unknown etiology. Ferulago angulata (Schltdl.) Boiss. has been proposed as an efficacious treatment for gastrointestinal ulcers and UC in the traditional Persian medicine. Objective: In the present study, we investigated the possible protective effect of F. angulata (Schltdl.) Boiss. hydroalcoholic extract (FAE) against acetic acid-induced UC in rats. Materials and Methods: The aerial parts of FAE (50, 100, and 200 mg/kg) were administered orally after acetic acid induced-inflammatory bowel disease. On the 3rd day, animals were euthanized by an overdose of ether inhalation and the abdomen was rapidly dissected for macroscopic, histological, and biochemical (tumor necrosis factor [TNF]-α, interleukin-1β [IL-1β]) analyses. Results: All doses of F. angulata (Schltdl.) Boiss. extract ameliorated acetic acid-induced macroscopic and microscopic scores, except for 50 mg/kg. Furthermore, our observations were accompanied by a significant (P < 0.05) suppression in the elevated amount of IL-1β and TNF-α. Conclusion: These findings demonstrated the beneficial effect of hydroalcoholic extract of F. angulata (Schltdl.) Boiss. in acetic acid-induced ulcerative lesions via anti-inflammatory, immunomodulatory, and mucosal healing properties. F. angulata (Schltdl.) Boiss. possesses promising protective function on colitis. Abbreviations Used: IBD: Inflammatory bowel disease; UC: Ulcerative colitis; CD: Crohn's disease; TNF-α: Tumor necrosis factor-α; IL-1β: Interleukin-1β; PMN: Polymorphonuclear; NO: Nitric oxide; FAE: Ferulago angulata (Schltdl.) Boiss. hydroalcoholic extract; COX: Cyclooxygenase.
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In vitro assessment of antioxidant, antimicrobial, cytotoxic, anti-inflammatory, and antidiabetic activities of Campanula retrorsa crude extracts p. 397
Josiane Alhage, Hoda Elbitar, Samir Taha, Thierry Benvegnu
DOI:10.4103/pr.pr_73_18  
Background: Campanula retrorsa belongs to the Campanulaceae family and the Campanula genus. The species of this latter are traditionally known for their antiallergic, spasmolytic, antiphlogistic, antioxidant, and antiviral properties. Few studies were realized on the bioactivity of Calicotome villosa crude extracts. Objective: The objective was to investigate, for the first time, the antioxidant, antimicrobial, cytotoxic, antidiabetic, and anti-inflammatory potentials of dichloromethane, methanol, and aqueous crude extracts of C. retrorsa leaves, flowers, and stems. Materials and Methods: The antioxidant activity of different crude extracts was determined using the free radical, 1, 1-diphenyl-2-picrylhydrazyl. Antimicrobial activity against three Gram-positive (Staphylococcus aureus, Bacillus cereus, and Enterococcus faecium), two Gram-negative (Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumanii) bacterial strains and a fungal strain (Candida albicans) was evaluated in a plate-hole diffusion assay. Cytotoxic activity of C. retrorsa dichloromethane and methanol crude extracts on Hela cells was realized using the Trypan blue dye exclusion technique. Antidiabetic activity against α-glucosidase and anti-inflammatory activity against bee venom phospholipase A2 (PLA2) were assessed using a colorimetric test. Results: Leaf and flower aqueous extracts possessed very high radical scavenging activity = 90%. The dichloromethane extracts of leaves and flowers were the most active against A. baumanii and C. albicans. After 48 h of treatment, all extracts except the methanol flower extract showed an inhibition of 90% of cell growth at 2 mg/mL. Dichloromethane and methanol stem crude extracts showed a modest inhibition of α-glucosidase. While no inhibition of the PLA2 was displayed by any of the tested extracts. Conclusion: Obtained results give an overall view on C. retrorsa crude extracts' bioactivities. Abbreviations Used: PLA2: Phospholipase A2; DPPH: 2, 2-Diphenyl-1-picrylhydrazy; CFU: Colony-forming unit; DMSO: Dimethyl sulfoxide; IZ: Inhibition zone; DMEM: Dulbecco Modified Eagle Medium; NaCl: Sodium Chloride; NaOH: Sodium Hydroxide; U: Enzyme Unit; CMUL: Microbiological Collection of the Lebanese University; ATCC: American Type Culture Collection; EC: Enzyme Commission number; CH2Cl2: Dichloromethane; CH3OH: Methanol; H2O: Water.
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Seabuckthorn (Hippophae rhamnoides and Hippophae salicifolia) seed oil in combating inflammation: A mechanistic approach p. 404
Suchita Dubey, MV Ramana, Anuradha Mishra, Pushpraj S Gupta, Himani Awasthi
DOI:10.4103/pr.pr_71_18  
Objective: This study assessed and compared in vivo anti-inflammatory activity of Hippophae rhamnoides (HR) and Hippophae salicifolia (HS) seed oil. Materials and Methods: HR and HS seed oil was extracted by Soxhlet apparatus and characterized using gas chromatography mass spectroscopy. Wistar rats were used for predicting anti-inflammatory activity. Results: HR and HS (2 and 4 ml/kg, respectively) exhibited dose-dependent inhibition of carrageenan-, histamine-, prostaglandin-, bradykinin-, and arachidonic acid (AA)-induced paw edema. Significant leukotriene-induced inhibition was observed in HR. Myeloperoxidase (MPO) and lipid-peroxidase (LPO) assays were performed and HR and HS seed oil significantly decreased the level of MPO and LPO at 4 ml/kg dose (P < 0.001). Conclusion: Dual inhibition of AA metabolism in HR and cyclooxygenase inhibition in HS was observed that might be attributed to the presence of polyunsaturated fatty acids (PUFAs), specifically, a correct balance of n-3 and n-6 PUFAs. However, the findings should be interpreted in the light of limitation of this study. Detailed experimentation at enzymatic levels would further help in substantiating the results inferred in this study. Abbreviations Used: AA: Arachidonic acid, ALA: Alpha linolenic acid, COX: Cyclooxygenase, EPA: Eicosapentaenoic acid, GCMS: Gas chromatography-mass spectroscopy, HR: Hippophae rhamnoides, HS: Hippophae salicifolia, ip: Intraperitoneal, LOX: Lipoxygenase, LPO: Lipid peroxidase, LTB4: Leukotriene B4, LTs: Leukotrienes, MPO: Myeloperoxidase, PGE2: Prostaglandin E2, PGs: Prostaglandins, PUFA: Poly unsaturated fatty acid, SBT: Seabuckthorn
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Evaluation of toxicological, diuretic, and laxative properties of ethanol extract from Macrothelypteris Torresiana (Gaudich) aerial parts with In silico docking studies of polyphenolic compounds on carbonic anhydrase II: An enzyme target for diuretic activity p. 408
Sumanta Mondal, Naresh Panigrahi, Purab Sancheti, Ruchi Tirkey, Prasenjit Mondal, Sara Almas, Venu Kola
DOI:10.4103/pr.pr_16_18  
Background: Macrothelypteris torresiana (Gaudich) is a species of fern having a wide range of reputed medicinal properties for the treatment of inflammation, fever, renal failure, stomach problems, etc. Objective: The present investigation focused on the evaluation of toxicity profile and diuretic and laxative activities of ethanol extract from M. torresiana aerial parts (EEMTAP), with in silico docking studies of polyphenolic compounds on carbonic anhydrase (CA)-II, an enzyme target for diuretic activity. Materials and Methods: Acute and subacute toxicity was performed according to the Organization for Economic Co-operation and Development guidelines. EEMTAP at doses of 200, 400, and 600 mg/kg, p.o., employed for the assessment of diuretic and laxative activities with loperamide-induced constipation in Wistar albino rats. Furosemide (10 mg/kg, p.o.), agar-agar (300 mg/kg, p.o.), and sodium picosulfate (5 mg/kg, p.o) were used as reference standards, respectively, for activity comparison. During saluretic activity study, total urine volume, body weight before and after the experiment, and urinary levels of Na+, K+ (by flame photometry), and Cl (by titrimetry) were estimated. Polyphenolic compounds such as caffeic acid and quercetin were successfully detected through chromatographic method of EEMTAP, and to rationalize the results obtained in diuretic activities, we carried out docking studies of the natural phenolic compounds against CA-II enzyme co-complexed with furosemide (Protein Data Bank ID: 1Z9Y CA-II in complex with furosemide as sulfonamide inhibitor). Results: In acute toxicity study, no mortality was observed at 2000 mg/kg, p.o., and in subacute toxicity study, the extract-treated group did not show any significant changes in body weight and organ weights. The hematological and biochemical parameters did not show any significant changes in the sample-treated groups when compared with the control group animals. The laxative activity of the extract was found to be in a dose-dependent increase in fecal output of rats at selected dose levels; similarly, EEMTAP significantly increased the urinary output as well as urinary electrolyte concentration in a dose-dependent manner. The molecular docking studies of phenolic compounds (caffeic acid and quercetin) into the binding site of CA II enzyme reveals that these analogues are having more favourable interaction when compared to the furosemide with better docking scores and hydrogen bonding interactions. Conclusion: The result demonstrated that the EEMTAP possesses a reasonable safety profile and shows promising diuretic and laxative activities in a dose-dependent manner. Abbreviations Used: EEMTAP: Ethanol Extract from M. Torresiana Aerial Parts; CA-I: Carbonic Anhydrase I; CA-II: Carbonic Anhydrase II; hCA-II: Human Carbonic Anhydrase-II; OECD: Organisation for Economic Co-operation and Development; PDB: Protein Data Bank.
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In vitro assessment of antioxidant and antiurolithic activities of ethanol extract of whole plant Biophytum sensitivum (Linn.) DC p. 417
BR Abhirama, Shanmugasundaram Rajagopal, IS Neethu, SG Ariya, B Athira, KM Dhanalekshmi
DOI:10.4103/pr.pr_30_18  
Background: Previous research studies have demonstrated that the formation of urinary stones leads to oxidative stress in patients; hence, search for antiurolithic drugs possessing antioxidant activities from natural sources has gained great potential. Objectives: The present study was undertaken to investigate in vitro antioxidant and antiurolithic potency of ethanol extract of whole plant Biophytum sensitivum Linn. DC (EEBS). Materials and Methods: The antioxidant potential of EEBS was determined by nitric oxide radical scavenging assay. Inhibition capacity of EEBS on calcium oxalate (CaOx) crystallization was evaluated by nucleation assay, aggregation assay, and microscopic assay (image analysis of CaOx crystal morphology). Results: Half-maximal inhibitory concentration (IC50) value of nitric oxide radical scavenging activity of EEBS was found to be 90.12 μg/mL and that of ascorbic acid (standard) was 37.23 μg/mL. In nucleation assay, IC50of EEBS was found to be 68.82 mg/mL, compared with 52. 41 mg/mL for cystone (standard). In aggregation assay, IC50value was indicated as 52.39 mg/mL and for cystone, it was found to be 41.62 mg/mL. Addition of various concentration of EEBS (20, 40, 80, and 160 mg/mL) resulted in change in structure of CaOx crystals. EEBS a concentration of 160 mg/mL reduced the size of CaOx crystals to 812.68 μm whereas size of CaOx crystal treated with the control was 1398.05 μm. The size reduction of CaOx crystals was found to be dose-dependent. Conclusion: This plant can be used alone or in combination with other herbal drugs, as EEBS showed significant antioxidant and antiurolithic activities. Abbreviations Used: EEBS: Ethanol extract of Biophytum sensitivum; CaOx: Calcium oxalate; IC50: Half-maximal inhibitory concentration; ESWL: Extracorporeal shockwave lithotripsy; Tris-HCl: Tris hydrochloride; NaCl: Sodium chloride; CaCl2: Calcium chloride; Na2C2O4: Sodium oxalate; COM: Calcium oxalate monohydrate; COD: Calcium oxalate dihydrate; Abs: Absorbance
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Validation of high-performance liquid chromatography method for quantification of costunolide content in Laurus nobilis p. 422
Puttaswamy Naveen, Harakanahalli Basavegowda Lingaraju, Siddappa Chandrappa, Koretekera Shekharappa Priya, Kodimule Shyam Prasad
DOI:10.4103/pr.pr_31_18  
Objective: To establish a method to determine the content of costunolide in Laurus nobilis leaves by high-performance liquid chromatography (HPLC). Materials and Methods: The separation was performed on a reversed-phase C18 column (100 Å, 150 mm × 4.6 mm, 5 μm pore size) using a mobile phase composed of water:acetonitrile (40:60) at a flow rate of 1.0 ml/min. The detection was carried out on a ultraviolet detector at 210 nm. The developed method was validated according to the requirements for International Conference on Harmonisation guidelines. Results: The proposed method for costunolide was validated for linearity with excellent correlation coefficient (r2 > 0.999). The relative standard deviation (RSD) is less than 1% in precision (i.e repeatability and intermediate) of the method. The recovery rate for costunolide was within 100.54%–102.62%. The limit of detection and limit of quantitation were 2.29 and 6.64 parts per million, respectively. Conclusion: The developed HPLC method is simple, rapid, precisely, accurately, and widely accepted and it is recommended for efficient assays in routine work. Abbreviation Used: HPLC: High-performance liquid chromatography; UV: Ultraviolet; ICH: International Conference on Harmonisation; r2: Correlation coefficient; WHO: World Health Organization; ppm: Parts per million; % RSD: Percentage relative standard deviation
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Oral intake of polyphenols of Chromolaena odorata: A perspective in peptic ulcer, thrombocytopenia, and heparin-induced bleeding diathesis in rodent model p. 426
Tania S Paul, Biswadeep B Das, Shakti P Ingale, Niruta Killedar, Kishori G Apte
DOI:10.4103/pr.pr_107_18  
Objective: Bleeding disorder is of two types: external bleeding due to trauma, surgery, and internal bleeding which happens mainly due to ulcers and thrombocytopenia or use of excessive anticoagulants. Proven hemostatic activity of Chromolaena odorata (L) in bleeding and wound healing had encouraged to evaluate the efficacy of aqueous extract and spray-dried powder (SDP) on ulcers, thrombocytopenia, and heparin-induced bleeding diathesis. Materials and Methods: Internal bleeding in the form of lesions in empty stomach was induced by absolute ethanol. Busulfan is administered through intraperitoneal injection to produce thrombocytopenia in Wistar rats and heparin at 1000 IU/ml was injected intravenous to mice, and after 30 min, tail was truncated to qualitatively and quantitatively differentiate the bleeding diathesis in treated versus nontreated controls. Results: A significant reduction in the lesions was observed in treated rats as compared to disease control rats which was evident from ulcer index and histopathology data. Considerable protection and production of platelets were observed in the thrombocytopenia model which was drastically reduced in the Busulfan administered disease control. Both aqueous extract and SDP were successful in preventing blood loss in heparin-induced tail-bleeding model. Conclusion: The presence of polyphenols attributes to the crucial role of C. odorata in arresting internal bleeding from stomach ulcer, protecting the thrombocytes from destroying, and minimizing the bleeding diathesis in mice. Abbreviations Used: SDP: Spray-dried powder, μL: Microliter, ml: Milliliter, nm: Nanometer, μg: Microgram, NAD: No abnormality detected, SD: Standard deviation.
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The study of safety and skin whitening efficacy of melinjo (Gnetum gnemon L.) seed extract-loaded lipid particle gel p. 432
Imalia Nurrachma Ayuningtyas, Abdul Mun'im, Sutriyo Sutriyo
DOI:10.4103/pr.pr_17_18  
Background: Melinjo (Gnetum gnemon L.) seed extract (MSE) is potential as skin-whitening agent because it contains trans-resveratrol and its derivatives, to inhibit tyrosinase in melanogenesis process. Using MSE in cosmetic products will be challenging due to resveratrol chemical instability and bioavailability in the skin. Many cosmetic products have been developed using lipid particle technology to improve their limitation. The objective of this research was to examine the skin safety and whitening efficacy of MSE-loaded lipid particle gel in healthy human subjects. Materials and Methods: Single occlusive closed patch test for 24 h was used as the skin irritation analysis. Irritation responses were graded after patch removal and compared to the control for evaluation. The efficacy study was performed using Mexameter to measure skin melanin index on 25 female volunteers. Results: The result showed the test product did not induce skin irritation effect. The skin melanin index was statistically significant decreased (P < 0.05) after 28 days of application the test product, with the averaged by 3.50%, and skin melanin index changed by increase 0.75% in the control group. Conclusion: Application MSE-loaded lipid particle gel can brighten the skin, without cause irritation under normal conditions of use. Abbreviations Used: α-MSH: α-melanocyte stimulating hormone; MITF: Microphthalmia-associated transcription factor; MSE: Melinjo seed extract.
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Effect of capsaicin on pharmacodynamic and pharmacokinetics of gliclazide in animal models with diabetes p. 437
Umachandar Lagisetty, Habibuddin Mohammed, Sivakumar Ramaiah
DOI:10.4103/pr.pr_81_18  
Background: Food–drug interactions can have great effect on the adverse effects of many drugs and also on the success of drug treatment. Gliclazide is one of the drugs of choice to treat type 2 diabetes. Capsaicin is present in Capsicum annuum, a spice found in regular food. Objective: The objective of the present study was to find out the pharmacodynamic (PD) and pharmacokinetic (PK) interactions of capsaicin on gliclazide in animal models using rats and rabbits. Materials and Methods: Single- and multiple-dose interaction studies were conducted in rats (normal and diabetic) and rabbits (diabetic) to evaluate the effect of capsaicin on the activity of gliclazide. Blood samples collected at predetermined time intervals from the experimental animals were used for the estimation of glucose and insulin levels using an automated clinical chemistry analyzer and radioimmunoassay method, respectively. β-cell function was determined by homeostasis model assessment. In addition, high-performance liquid chromatography technique was used to analyze the serum gliclazide levels in rabbits. Results: Capsaicin did not exhibit any hypoglycemic activity in normal rats, but exhibited significant antihyperglycemic activity in both diabetic rats and rabbits with improvement in insulin levels and β-cell function. Gliclazide showed significant reduction in blood glucose levels in normal and diabetic rats and diabetic rabbits. In addition, it significantly increased insulin levels and β-cell function in diabetic animals. The samples analyzed from all time points in combination with capsaicin showed significantly greater reduction in blood glucose levels and a significant increase in insulin levels and β-cell function in diabetic rats and rabbits. The PK parameters of gliclazide were also altered by capsaicin treatment in rabbits. Conclusion: The present study concluded that the interaction of capsaicin with gliclazide on single- and multiple-dose treatment was both PD and PK in nature. Abbreviations Used: CYP: Cytochrome P450, g: Gram, kg: Kilogram, C: Degree Celsius, %: Percentage, h: Hours, p.o.: Per oral, dL: Deciliter, μ: Micro, IU: International units, mL: milliliter, Cmax: Maximum concentration, Tmax: time to maximum, AUC: Area under the curve, AUMC: Area under the first moment curve, t1/2: Elimination half-life, kel: Elimination rate constant, MRT: Mean residence time, Cl: Clearance, ng: Nanogram, mg: Microgram
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