Antioxidant activity and free radical-scavenging capacity of Gynura divaricata leaf extracts at different temperatures

Chunpeng Wan, Yanying Yu, Shouran Zhou, Wei Liu, Shuge Tian, Shuwen Cao

Pharmacognosy Magazine 2011 7(25):40-45

Background: Extraction temperature influences the total phenolic content (TPC), total flavonoid content (TFC) of medicinal plant extracts to a great extend. TPC and TFC are the principle activity constituents present in the plant. The effects of extraction temperature on TPC, TFC and free radical-scavenging capacity of Gynura divaricata leaf extracts are worth to study. Materials and Methods: Folin-Ciocalteu and aluminum chloride colorimetric assay were used to determine the TPC and TFC of Gynura divaricata leaf extracts at different temperatures. The antioxidant and free radical-scavenging activity were measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) and phosphomolybdenum methods. Results: TPC and TFC were significantly elevated with increasing extraction temperature (from 40°C to 100°C). However, TPC and TFC were not significantly different (P > 0.05) at the extraction temperatures 90°C and 100°C. Also, the extracts obtained at a higher temperature exhibited a significant free radical-scavenging activity compared with extraction at lower temperatures (P < 0.05). The TPCs (13.95-36.68 mg gallic acid equivalent/g dry material) were highly correlated with DPPH (R2 = 0.9229), ABTS (R2 = 0.9951) free radical-scavenging capacity, and total antioxidant activity (R2 = 0.9872) evaluated by phosphomolybdenum method. Conclusion: The TPC and TFC of G. divaricata leaf was significantly influenced by the extraction temperatures, which were the main antioxidant constituents present in the G. divaricata plant.

The antihypertensive effect of ethyl acetate extract from red raspberry fruit in hypertensive rats

Han Jia, Ji Wen Liu, Halmurat Ufur, Geng Sheng He, Hai Liqian, Peipei Chen

Pharmacognosy Magazine 2011 7(25):19-24

Objectives: To evaluate the antihypertensive effect of Xinjiang red raspberry fruit ethyl acetate extract (EER) on spontaneously hypertensive rats (SHR) and its possible mechanism from antioxidant perspective. Materials and Methods: The SHR rats were randomly divided into 3 groups, and treated with EER low dose (EERL, 100 mg/kg/d), high dose (EERH, 200 mg/kg/d), and water (SHR) through gastric gavage daily for 5 weeks. Another 8 age-matched male Wistar-Kyoto rats were used as normotensive group (WKY). The systolic blood pressure (SBP) was measured by noninvasive tail-cuff method once a week. At the end of the treatment, blood samples were collected and serum concentrations of nitric oxide (NO), superoxide dismutase (SOD), malondialchehyche (MDA), and plasma endothelin (ET) were determined. Results: Treatment of SHR rats with EER lowered the blood pressure compared with that treated with water (SHR), and the high dose showed more significant reduction in blood pressure. Treatment of SHR rats with EER increased serum NO and SOD levels and lowered ET and MDA levels. As compared with control group, NO levels were increased significantly in EERL (P < 0.01), SOD was elevated more significantly in both EERL and EERH (P < 0.01); MDA was decreased significantly in EERH group (P < 0.05), whereas plasma ET decreased more significantly in the EERH group (P < 0.05). Conclusions: The red raspberry extracts demonstrated a dose-dependent antihypertensive effects in SHR and this may be related to increased NO activation and improved vascular endothelial dysfunction via antioxidation. These results confirmed that raspberries rich in polyphenols have potential cardiovascular protective effects.

Pharmacovigilance of herbal medicines: Current state and future directions

Sandeep Shetti, C Dinesh Kumar, Neeraj Kumar Sriwastava, Indra Prakash Sharma

Pharmacognosy Magazine 2011 7(25):69-73

Currently, a majority of the adverse events related to the use of herbal products and herbal medicines that are reported are attributable either to poor product quality or to improper use. Inadequate regulatory measures, weak quality control systems, and largely uncontrolled distribution channels (including mail order and Internet sales) may have been contributing to the occurrence of such events. In order to expand the knowledge about genuine adverse reactions to herbal medicines, and to avoid wasting scarce resources for identifying and analyzing adverse events, events resulting from such situations will need to be reduced or eliminated. Member States of the World Health Organization (WHO) are therefore encouraged to strengthen national regulation, registration and quality assurance and control of herbal medicines. In addition, the national health authorities should give greater attention to consumer education and to qualified practice in the provision of herbal medicines.

The relaxant effect induced by Allium sativum L. bulb aqueous extract on rat isolated trachea

Badreddine Fehri, KK Mueen Ahmed, Jean-Marc Aiache

Pharmacognosy Magazine 2011 7(25):14-18

Background: Garlic plays an important role in complementary and alternative medicine. Most people believe in and use herbal products even when they have not been as thoroughly researched as garlic. Garlic is also known for its beneficial effects on the cardiovascular system. Materials and Methods: The relaxant effect of Allium sativum L. bulb aqueous extract (ASBAE) containing 0.06%-0.10% of allicin was studied on isolated smooth muscle of trachea of rats precontracted using acetylcholine (10−5 M). Results: It was found that ASBAE induced a dose-dependent relaxation with recorded EC 50 values of 71.87 ± 5.90 μg/mL (n = 7). Pretreatments with mepyramine (10−7 M), methysergide (10−7 M), caffeine (10−6 M), theophylline (10−6 M), nifedipine (10−6 M), and dipyridamole (10−6 M) did not alter ASBAE concentration-response curves. In turn, concentration-response curves to ASBAE were significantly shifted toward right in the presence of aspirin (3.10−3 M), indomethacin (10−6 M), prazosin (10−6 M), and propranolol (10−7 M). Conclusion: It is suggested that the recorded relaxation results are due to the release of prostaglandins E 1 and E 2 consecutively to a- and b-adrenoreceptor stimulation.

Honey induces apoptosis in renal cell carcinoma

Saeed Samarghandian, Jalil Tavakkol Afshari, Saiedeh Davoodi

Pharmacognosy Magazine 2011 7(25):46-52

Background: The fact that antioxidants have several preventative effects against different diseases, such as coronary diseases, inflammatory disorders, neurologic degeneration, aging, and cancer, has led to the search for food rich in antioxidants. Honey has been used as a traditional food and medical source since ancient times. However, recently many scientists have been concentrating on the antioxidant property of honey. By use of human renal cancer cell lines (ACHN), we investigated the antiproliferative activity, apoptosis, and the antitumor activity of honey. Materials and Methods: The cells were cultured in Dulbecco's modified Eagle's medium with 10% fetal bovine serum treated with different concentrations of honey for 3 consecutive days. Cell viability was quantitated by the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptotic cells were determined using Annexin-V-fluorescein isothiocyanate (FITC) by flow cytometry. Results: Honey decreased the cell viability in the malignant cells in a concentration- and time-dependent manner. The IC 50 values against the ACHN cell lines were determined as 1.7 ± 0.04% and 2.1 ± 0.03% μg/mL after 48 and 72 h, respectively. Honey induced apoptosis of the ACHN cells in a concentration-dependent manner, as determined by flow cytometry histogram of treated cells. Conclusion: It might be concluded that honey may cause cell death in the ACHN cells, in which apoptosis plays an important role. Most of the drugs used in the cancer treatment are apoptotic inducers, hence apoptotic nature of honey is considered vital. Therefore, it prompted us to investigate honey as a potential candidate for renal cancer treatment.

A HPLC-DAD method for the simultaneous determination of five marker components in the traditional herbal medicine Bangpungtongsung-san

Jin Bae Weon, Hye Jin Yang, Jin Yeul Ma, Choong Je Ma

Pharmacognosy Magazine 2011 7(25):60-64

Background: Bangpungtongsung-san, one of the traditional herbal medicines, was known to be a prescription for obesity. Objective: For the simultaneous determination of five components (paeoniflorin, 6-gingerol, decursin, geniposide, and glycyrrhizin) in Bangpungtongsung-san, a high-performance liquid chromatography with diode-array detector method was established. Materials and Methods: To develop the method, a reverse phase column, DIONEX C 18 (5 μm, 120 Å, 4.6 mm × 150 mm) was used. The mobile phase consisted of methanol and water using a gradient elution. The UV wavelength was set at 230, 240, and 254 nm. Method validation was accomplished by linearity, precision test, and recovery test. Results: All calibration curves of components showed good linearity (R 2 > 0.9959). The limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.01 to 0.17 μg/ml and 0.04 to 0.53 μg/ml, respectively. The relative standard deviations (RSD) value of precision test, intraday and interday tests were less than 0.43% and 1.26%. In the recovery test, results of accuracy ranged from 95.27% to 107.70% with RSD values less than 2.21%. Conclusion: This developed method was applied to the commercial Bangpungtongsung-san sample and the five marker components were separated effectively without interference of any peaks of components.

Orthogonal test design for optimization of the extraction of polysaccharide from Paeonia sinjiangensis K.Y. Pan

Shuge Tian, Xiaoying Zhou, Haiyan Gong, Xiuming Ma, Fan Zhang

Pharmacognosy Magazine 2011 7(25):4-8

Background: Paeonia sinjiangensis K.Y. Pan is a perennial herb belonging to the ranunculaceae family and it is one of the most important crude drugs in traditional Chinese medicine. In this article, Paeonia sinjiangensis K.Y. Pan rich in polysaccharide is used as an experimental material. Materials and Method: Study the effects of proportion, temperature, times and time taken for the extraction yield of polysaccharide through a single-factor exploration. Then, through an orthogonal experiment (L9(3)4), it was investigated to get the best extraction conditions. Results: The results showed that the ratio of solvent to raw material, number of extractions and duration of extraction were the main variables that influenced the yields of extracts. The separation procedure of precipitation with alcohol and the purification from the removing proteins were deeply analyzed. Meanwhile the contents of polysaccharide were determined by anthrone colorimetry. Conclusion: The highest yield was obtained when the ratio of solvent to raw material, number of extractions, and duration of extraction were 8:1, 2, and 1.5 h, respectively. The content of soluble polysaccharide is 51.57%.

Modulating effect of Gmelina arborea Linn. on immunosuppressed albino rats

SH Shukla, AK Saluja, SS Pandya

Pharmacognosy Research 2010 2(6):359-363

Aim: In the present study, the immunomodulatory effects of roots of Gmelina arborea Linn. were investigated. Materials and Methods: Methanolic extract of G. arborea Linn. (MEGA) and its ethyl acetate fraction (EAFME) were used for evaluating the pharmacological activity. The modulating effect was evaluated on humoral and cell-mediated immune response using animal models like cyclophosphamide-induced myelosuppression, delayed-type hypersensitivity (DTH) response, and humoral antibody (HA) titre. Results: Both test extracts produced significant increase in HA titre, DTH response, and levels of total white blood cell count. Conclusion: This drug is found to be a potential immunostimulant.

An automated or semi-automated identification system using venation pattern to delimit Indian leaf drugs: A proposal

A.B.D. Selvam

Pharmacognosy Research 2010 2(6):385-387

About 7,200 medicinal plants are known to occur in India, of which, the leaves of a few hundred plants have medicinal properties. Identification of leaf drugs using venation is considered as one of the most reliable and convenient methods. Leaf identification by mechanical means may often lead to wrong identification. Due to the growing volume of illegal trade/malpractice in the crude drug industry on the one hand and lack of sufficient experts on the other, a much faster, convenient and reliable method is mandatory for the identification of Indian leaf drugs. Therefore, a new automated or semi-automated identification system based on venation pattern is inevitable for the present day condition to identify and authenticate the leaves of Indian medicinal plants.

Evaluation of in vitro cytotoxic effect of Trichosanthes dioica root

Sanjib Bhattacharya, Pallab K Haldar

Pharmacognosy Research 2010 2(6):355-358

Background: Trichosanthes dioica Roxb. (Cucurbitaceae), called pointed gourd in English is a dioecious climber grown in India and used traditionally for various medicinal purposes. Methods: Present study was aimed to evaluate in vitro cytotoxic effect of dichloromethane (DCTD), methanol (METD), and aqueous (AQTD) extracts of T. dioica root using Allium cepa root meristems by keeping them in different concentrations of each test extract under specific experimental conditions followed by determination of root growth inhibition (root length and number) and mitotic index. Results: All the extracts significantly demonstrated concentration-dependent inhibition of root length and number and reduction in mitotic index, indicating antimitotic activity demonstrating cytotoxicity and genotoxicity. DCTD was found to be the most potent (EC 50 : 2.8 mg/ml), followed by METD and AQTD. Conclusion: The present study therefore, establishes promising in vitro cytotoxic and genotoxic property of T. dioica root against the test system.

Influence of sonication on the phenolic content and antioxidant activity of Terminalia catappa L. leaves

HV Annegowda, LN Anwar, MN Mordi, S Ramanathan, SM Mansor

Pharmacognosy Research 2010 2(6):368-373

Aim: This study was designed to evaluate the phenolic content and antioxidant activity of ethanolic extracts from T. catappa leaves obtained by different intervals of sonication. Methods: Three commonly used methods were followed to evaluate phenolic content and four in vitro methods like 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging assay, ferric reducing antioxidant potency (FRAP), and total antioxidant capacity assays for measuring the antioxidant activities. Antioxidant values of these assays were expressed in terms of milligrams vitamin C equivalent (VCE) antioxidant activities. Results: This study showed that extract obtained with 40 minutes of sonication possessed significant (P < 0.05) polyphenolic contents compared to 20 and 60 minutes sonication and control (24 hour maceration). Moreover, sonication of T. catappa leaf above 40 minutes was found to be unsuitable for extracting out phenolic contents. Even the results of antioxidant assays showed that 40 minutes of the sonicated extract exhibited significant (P < 0.05) VCE values compared to extracts obtained at different intervals of sonication and control. Conclusions: In sonication extraction method 40 minutes is an ideal time to obtain extract enriched with high polyphenolic content with good antioxidant activity from T. catappa leaves.

Possible implication of oxidative stress in anti filarial effect of certain traditionally used medicinal plants in vitro against Brugia malayi microfilariae

RD Sharma, AR Veerpathran, G Dakshinamoorthy, KN Sahare, K Goswami, M.V.R. Reddy

Pharmacognosy Research 2010 2(6):350-354

Introduction: Tropical disease research scheme of World Health Organization has duly recognized traditional medicine as alternative for antifilarial drug development. Polyphenolic compounds present in traditionally used herbal medicines are natural antioxidants; however, paradoxically they may exert pro-oxidant effect. Popular drug diethyl carbamazine citrate harnesses the innate inflammatory response and the consequent oxidative assault to combat invading microbes. Methods: With this perspective, extracts of Vitex negundo L. (roots), Butea monosperma L. (leaves), Aegle marmelos Corr. (leaves), and Ricinus communis L. (leaves) were selected to explore the possible role of oxidative rationale in the antifilarial effect in vitro. Results: Apart from the last, other three plant extracts were reported to have polyphenolic compounds. Dose-dependent increase was found in the levels of lipid peroxidation and protein carbonylation for all the three plant extracts except Ricinus communis L. (leaves). Such increase in oxidative parameters also showed some degree of plant-specific predilection in terms of relatively higher level of particular oxidative parameter. High degree of correlation was observed between the antifilarial effect and the levels of corresponding oxidative stress parameters for these three plants. However, extracts of Ricinus communis L. (leaves) which is relatively deficient in polyphenolic ingredients recorded maximum 30% loss of motility and also did not show any significant difference in various stress parameters from corresponding control levels. Conclusion: These results reveal that targeted oxidative stress might be crucial in the pharmacodynamics.

Secondary metabolites and bioactivities of Myrtus communis

Mahmoud I Nassar, El-Sayed A Aboutabl, Rania F Ahmed, Ezzel-Din A El-Khrisy, Khaled M Ibrahim, Amany A Sleem

Pharmacognosy Research 2010 2(6):325-329

Background: Myrtus species are characterized by the presence of phenolic acids, flavonoids, tannins, volatile oils and fatty acids. They are remedies for variety of ailments. This study therefore investigated medicinal effects of Myrtus communis L. Methods: Bioactivity studies of Myrtus communis L. leaves were carried out on volatile oil, 7% methanol and aqueous extracts and the isolated compounds myricetin 3-O-β-glucopyranoside, myricetin 3-O-∝-rhamnopyranoside and gallic acid. Results: Determination of the median lethal dose (LD 50 ) revealed that the volatile oil, alcoholic and aqueous extracts were practically nontoxic and highly safe as no lethality was observed. The tested materials (volatile oil, alcoholic and aqueous extracts, myricetin 3-O-β-glucopyranoside, myricetin 3-O-∝-rhamnopyranoside and gallic acid) showed significant antihyperglycemic, anti-inflammatory and antinociceptive effects as compared with control groups and reference drugs. Conclusion : Administration of extracts of M. communis leaves could be safe at the dose used in this study.

Isolation and characterization of phytoconstituents from Chlorophytum borivilianum

Sharada L Deore, Somshekhar S Khadabadi

Pharmacognosy Research 2010 2(6):343-349

Background: The present communication deals with the identification and characterization of bioactive principles from the roots of Chlorophytum borivilianum. Method: Methanolic extract and its fractions were used to isolate different phytoconstituents. The structures of isolated compounds were characterized and elucidated with chemical and spectroscopic techniques such as Infra Red, Nuclear Mass Resonace and Mass spectroscopy experiments. Fatty acids were characterized by GC-MS analysis. Result: Three Fatty acids were isolated and confirmed. One sterol stigmasterol was isolated. One new saponin named as Chlorophytoside-I (3b, 5a, 22R, 25R)-26-(β-D-glucopyranosyloxy)-22-hydroxy-furostan-12-one-3 yl O-β-D-galactopyranosyl (1-4) glucopyranoside was isolated. Conclusion: The roots of Chlorophytum borivilianum contain three important fatty acids, common sterol stigmasterol and one furostanol saponins.

A comparative profile of methanol extracts of Allium cepa and Allium sativum in diabetic neuropathy in mice

Abhishek Bhanot, Richa Shri

Pharmacognosy Research 2010 2(6):374-384

Introduction: Diabetic Neuropathy (DN) is a major microvascular complication of uncontrolled diabetes. This may result from increased oxidative stress that accompanies diabetes. Hence plants with antioxidant action play an important role in management of diabetes and its complications. Materials and Methods: This study was designed to evaluate preventive as well as curative effect of methanol extracts of outer scales and edible portions of two plants with established antioxidant action – Allium cepa and Allium sativum, in induced DN in albino mice. Mice were divided into control, diabetic and test extracts treated groups. Test extracts were administered daily at a dose of 200 mg/kg p.o. for 21 days, in the preventive group prior to onset of DN, and in the curative group after the onset of DN. Hyperalgesia and oxidative stress markers were assessed. STZ-diabetic mice showed a significant thermal hyperalgesia (as assessed by the tail-flick test), indicating development of DN. Results: Treatment with test extracts prevented loss in body weight, decreased plasma glucose level, and significantly ameliorated the hyperalgesia, TBARS, serum nitrite and GSH levels in diabetic mice. Conclusion: Methanol extract of outer scales of onion has shown most significant improvement; may be due to higher content of phenolic compounds in outer scales of A. cepa.

A comparison of the cytotoxic potential of standardized aqueous and ethanolic extracts of a polyherbal mixture comprised of Nigella sativa (seeds), Hemidesmus indicus (roots) and Smilax glabra (rhizome)

Sameera R Samarakoon, Ira Thabrew, Prasanna B Galhena, Dilip De Silva, Kamani H Tennekoon

Pharmacognosy Research 2010 2(6):335-342

Background: A decoction (hot-water extract) comprised of Nigella sativa (seeds), Hemidesmus indicus (roots), and Smilax glabra (rhizome) has been reported to prevent chemically-induced hepatocarcinogenic changes in rats and to exert significant cytotoxic effects on human hepatoma (HepG2) cells. However, the decoction used in previous studies to determine cytotoxicity was not standardized. Further, during preparation of pharmaceuticals for clinical use, it is more convenient to use an ethanolic extract. Therefore this study was carried out to (a) develop standardized aqueous and ethanolic extracts of the plant mixture (N. sativa, H. indicus, and S. glabra) used in the preparation of the original decoction, and (b) compare the cytotoxic effects of these two extracts by evaluating cytotoxicity to the human hepatoma (HepG2) cell line. Methods: Aqueous and ethanolic extracts have been standardized by evaluating organoleptic characters, physicochemical properties, qualitative and quantitative analysis of chemical constituents, and analysis of High Performance Liquid Chromatography (HPLC) and Thin Layer Chromatography (TLC) profiles. Cytotoxic potentials of the above standardized extracts were compared by evaluating their effects on the survival and overall cell activity of HepG2 cells by use of the 3-(4, 5-dimethylthiazol-2yl) -2, 5 – biphenyl tetrazolium bromide (MTT) and Sulphorhodamine B (SRB) assays. Results: Results from MTT and SRB assays demonstrated that both extracts exerted strong dose-dependent in vitro cytotoxicity to HepG2 cells. The standardized aqueous extract showed a marginally (though significantly, P<0.05) higher cyotoxic potential than the ethanolic extract. Thymoquinone, an already known cytotoxic compound isolated from N. sativa seeds was only observed in the standardized ethanolic extract. Thus, compounds other than thymoquinone appear to mediate the cytotoxicity of the standardized aqueous extract of this poly-herbal preparation. Conclusion: It may be concluded that results obtained in the present study could be used as a diagnostic tool for the correct identification of these aqueous or ethanolic extracts and would be useful for the preparation of a standardized pharmaceutical product that may be used in the future for clinical therapy of hepatocellular carcinoma.

Inhibitory effect of rutin and curcumin on experimentally-induced calcium oxalate urolithiasis in rats

Jaydip Ghodasara, Anil Pawar, Chinmay Deshmukh, Bhanudas Kuchekar

Pharmacognosy Research 2010 2(6):388-392

Background: Renal epithelial cell injury by reactive oxygen species is pre-requisite step in the pathogenesis of urolithiasis. Rutin and curcumin are polyphenolic compounds known to have antioxidant and anti-inflammatory activities, but their effect on urolithiasis is yet to be elucidated. In the present study, we have investigated the inhibitory effect of rutin and curcumin on calcium oxalate urolithiasis in Wistar albino rats. Methods: Calcium oxalate urolithiasis was induced experimentally by administration of 0.75% v/v ethylene glycol with 1% w/v ammonium chloride in drinking water for three days followed by only 0.75% v/v ethylene glycol for 25 days. Rutin (20 mg/kg body weight) and curcumin (60 mg/kg body weight) were given once daily for 28 days by oral route. After treatment period, calcium and oxalate levels in urine and kidney tissue homogenate were measured. Kidney was also used for histopathological examination. Results: Stone-induction with ethylene glycol and ammonium chloride resulted in elevated levels of calcium and oxalate in the urine and kidney sample, whereas supplementation of rutin and curcumin restored it near to normal. Histopathological study revealed minimum tissue damage and less number of calcium oxalate deposits in kidney of animal treated with rutin and curcumin as compared to calculi-induced animal. Conclusion: The data suggest that the rutin and curcumin inhibits calcium oxalate urolithiasis. This effect is mediated possibly through a lowering of urinary concentration of stone forming constituents, anti-inflammatory and antioxidant effects.

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16171819202122
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