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   2013| October-December  | Volume 9 | Issue 36  
    Online since September 7, 2013

 
 
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ORIGINAL ARTICLES
Protective effects of onion-derived quercetin on glutamate-mediated hippocampal neuronal cell death
Eun-Ju Yang, Geum-Soog Kim, Jeong Ah Kim, Kyung-Sik Song
October-December 2013, 9(36):302-308
DOI:10.4103/0973-1296.117824  PMID:24124281
Background: Neurodegenerative diseases are characterized by progressive neuron degeneration in specific functional systems of the central or peripheral nervous system. This study investigated the protective effects of quercetin isolated from onion on neuronal cells and its protective mechanisms against glutamate-induced apoptosis in HT22 cells. Materials and Methods: HT22 cells were cultured to study the neuroprotective mechanism of quercetin against glutamate-mediated oxidative stress. The intracellular reactive oxygen species (ROS) level and mitochondrial membrane potential (ΔΨ m ) were measured. The protein expression of calpain, spectrin, Bcl-2, Bax, Bid, cytochrome c, and mitogen-activated protein kinases (MAPKs) was evaluated by Western blotting. Results: Quercetin had a protective effect by reducing both intracellular ROS overproduction and glutamate-mediated Ca 2+ influx. These effects were due to the downregulation of several apoptosis-related biochemical markers. Calpain expression was reduced and spectrin cleavage was inhibited by quercetin in glutamate-exposed HT22 cells. Disruption of the mitochondrial membrane potential (ΔΨ m ), activation of the pro-apoptotic proteins Bid and Bax, and cytochrome c release in response to glutamate-induced oxidative stress were reduced. Quercetin also suppressed phosphorylation of MAPKs. Conclusion: This is the first report on the detailed mechanisms of the protective effect of quercetin on HT22 cells. Onion extract and quercetin may be useful for preventing or treating neurodegenerative disorders.
  10 2,012 30
Optimization of enzyme-assisted extraction and characterization of collagen from Chinese sturgeon (Acipenser sturio Linnaeus) skin
Weiwei Feng, Ting Zhao, Ye Zhou, Fang Li, Ye Zou, Shiqi Bai, Wei Wang, Liuqing Yang, Xiangyang Wu
October 2013, 9(36):32-37
DOI:10.4103/0973-1296.117859  PMID:24143042
Background: Sturgeon (Acipenser sturio Linnaeus) skin contains high amount of nutrients including unsaturated fatty acids and collagen. A pepsin-assisted extraction procedure was developed and optimized for the extraction of collagen from Chinese sturgeon (Acipenser sturio Linnaeus) skins. Objective: To determine the optimum conditions with the maximum yield of the pepsin-soluble collagen (PSC) extraction. Materials and Methods: The conditions of the extraction were optimized using response surface methodology. The Box-Behnken design was used to evaluate the effects of the three independent variables (extraction time, enzyme concentration, and solid-liquid ratio) on the PSC yield of the sturgeon skin. Results: The optimal conditions were: solid-liquid ratio of 1:11.88, enzyme concentration of 2.42%, and extraction time of 6.45 h. The maximum yield of 86.69% of PSC was obtained under the optimal conditions. This value was not significantly different from the predicted value (87.4%) of the RSM (P < 0.05). Conclusion: The results of this study indicated that the production of PSC from sturgeon skin is feasible and beneficial. The patterns of sodium dodecyl sulfate-polyacrylamide gel electrophoretic patterns (SDS-PAGE) indicated that the sturgeon skin contains type I collagen, which is made of α-chain and β-chain. The infrared spectra of the collagens also indicated that pepsin hydrolysis does not affect the secondary structure of collagen, especially triple-helical structure.
  8 3,638 20
Modulation of drug efflux by aloe materials: An In Vitro investigation across rat intestinal tissue
Beneke Carien, Viljoen Alvaro, Hamman Josias
October 2013, 9(36):44-48
DOI:10.4103/0973-1296.117864  PMID:24143044
Background: Clinically, significant herb-drug interactions have been previously documented and can be pharmacodynamic and/or pharmacokinetic in nature. Pharmacokinetic interactions have been attributed to induction or inhibition of either metabolic enzymes or efflux transporters. Objective: The effect of gel and whole leaf materials from 3 different aloe species namely Aloe ferox, Aloe marlothii, and Aloe vera as well as polysaccharides precipitated from the A. vera materials on the bi-directional transport of cimetidine across rat intestinal tissue was investigated. Materials and Methods: Cimetidine transport studies were performed across excised rat intestinal tissue mounted in Sweetana-Grass diffusion chambers in both the apical-to-basolateral and basolateral-to-apical directions. Results: While A. vera gel and whole leaf materials did not inhibit the efflux of cimetidine, the polysaccharides precipitated from them did show a reduction of cimetidine efflux. On the other hand, both A. ferox and A. marlothii gel and whole leaf materials exhibited an inhibition effect on cimetidine efflux. Conclusions: This study identified a modulation effect of efflux transporters by certain aloe materials. This may cause herb-drug pharmacokinetic interactions when drugs that are substrates for these efflux transporters are taken simultaneously with aloe materials. On the other hand, these aloe materials may be used for drug absorption enhancement for drugs with low bioavailability due to extensive efflux.
  6 1,794 15
Cytoprotective and antioxidant effects of phenolic compounds from Haberlea rhodopensis Friv. (Gesneriaceae)
Magdalena Kondeva-Burdina, Dimitrina Zheleva-Dimitrova, Paraskev Nedialkov, Ulrich Girreser, Mitka Mitcheva
October-December 2013, 9(36):294-301
DOI:10.4103/0973-1296.117822  PMID:24124280
Background: Haberlea rhodopensis Friv. (Gesneriaceae) is a rare poikilohydric endemic and preglacial relict growing in Balkan Peninsula. Previous investigations demonstrated strong antioxidant, antimicrobial and antimutagenic potential of alcoholic extract from the plant. Objective: The isolation of known caffeoyl phenylethanoid glucoside - myconoside and flavone-C-glycosides hispidulin 8-C- (2-O-syringoyl-β-glucopyranoside), hispidulin 8-C-(6-O-acetyl-2-O-syringoyl-β-glucopyranoside), and hispidulin 8-C-(6-O-acetyl -β-glucopyranoside) from the leaves of H. rhodopensis was carried out. The aim of this study was to investigate cyto-protective and antioxidant effects of isolated compounds. Materials and Methods: Antioxidant activity of isolated substances was examined using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radicals; ferric reducing antioxidant power (FRAP) assay and inhibition of lipid peroxidation (LPO) in linoleic acid system by ferric thyocianate method. The compounds were investigated for their possible protective and antioxidant effects against tert -butyl hydroperoxide-induced oxidative stress in isolated rat hepatocytes. The levels of thiobarbituric acid reactive substances were assayed as an index of LPO. Lactate dehydrogenase leakage, cell viability, and reduced glutathione depletion were used as signs of cytotoxicity. Results: Myconoside demonstrated the highest DPPH radical scavenging, ABTS, FRAP, and antioxidant activity in linoleic acid system as well as the highest and statistically most significant protection and antioxidant activity against the toxic agent. Conclusion: Phenolic compounds isolated from H. rhodopensis demonstrated significant cytoprotective, radical scavenging potential, and inhibit lipid peroxidation, moreover, myconoside was found to be a new powerful natural antioxidant.
  6 2,317 20
Bioactivity studies on Musa seminifera Lour
Sanjib Saha, Jamil A Shilpi, Himangsu Mondal, Royhan Gofur, Morsaline Billah, Lutfun Nahar, Satyajit D Sarker
October-December 2013, 9(36):315-322
DOI:10.4103/0973-1296.117827  PMID:24124283
Background: Musa seminifera Lour is a tree-like perennial herb that has been used in folk medicine in Bangladesh to heal a number of ailments. Objective: To evaluate the antioxidant, analgesic, antidiarrheal, anthelmintic activities, and general toxicity of the ethanol extract of the roots. Materials and Methods: The extract was assessed for free-radical-scavenging activity by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, total phenolic content (TPC) by the Folin Ciocalteu reagent, antioxidant activity by the ferric reducing power assay, analgesic activity by the acetic acid-induced writhing and hot-plate tests, antidiarrheal activity by the castor oil-induced diarrhea model in mice, anthelmintic activity on Paramphistomum cervi and Haemonchus contortus, and general toxicity by the brine shrimp lethality assay. Results: The extract showed free-radical-scavenging activity with an IC 50 value of 44.86 μg/mL. TPC was 537.89 mg gallic acid equivalent/100 g of dried plant material. It showed concentration-dependent reducing power, and displayed 42.11 and 69.32% writhing inhibition at doses of 250 and 500 mg/kg body weight, respectively. The extract also significantly raised the pain threshold at the above-mentioned dose levels. In vivo antidiarrheal property was substantiated by significant prolongation of latent period and decrease in total number of stools compared with the control. The LC 50 against brine shrimp nauplii was 36.21 μg/mL. The extract exhibited dose-dependent decrease in paralysis and death time of the helminths. Conclusion: The above results demonstrated that the plant possesses notable bioactivities and somewhat supports its use in folk medicine.
  5 2,353 19
Mitigation of starch and glucose-induced postprandial glycemic excursion in rats by antioxidant-rich green-leafy vegetables' juice
Ashok Kumar Tiwari, Atmakuri Lakshmana Jyothi, Vasantharao Brahma Tejeswini, Kuncha Madhusudana, Domati Anand Kumar, Amtul Zehra, Sachin Bharat Agawane
October 2013, 9(36):66-73
DOI:10.4103/0973-1296.117872  PMID:24143048
Objective: Consumption of green-leafy vegetables is being advocated beneficial for type 2 diabetes mellitus individuals possibly because they are cost effective source of potent biological antioxidants. This research analyzed various phytochemicals, free radicals scavenging antioxidant potentials and starch digesting enzymes inhibitory activities in fresh juice of nine green-leafy vegetables. Furthermore, this study also investigated influence of these vegetables juice on starch and glucose induced postprandial glycemic load. Materials and Methods: Phytochemical constituents, in vitro free radicals scavenging antioxidant and enzymes inhibitory activities were evaluated applying various reported methods. Post-prandial glycemic excursion was induced in rats pretreated with vegetables juice by oral administration of starch and glucose. Results: All the leafy vegetables juice displayed potent free radicals scavenging activities. Juice of amaranthus, rumex, palak and raphanus displayed potential anti-oxidative property by reducing H 2 O 2 induced hemolysis in rats red blood cells RBCs. Ajwain and rumex juice showed pancreatic α-amylase inhibitory activity. Alternanthera, ajwain, methi, amaranthus and sowa leaves juice displayed intestinal α-glucosidase inhibitory activity. Juice of raphanus, ajwain and sowa significantly mitigated starch-induced postprandial glycemic load. Amaranthus leaves juice potently mitigated glucose-induced postprandial glycemic load and also reduced hemoglobin glycation induced by glucose in vitro. Conclusions: This investigation finds that juice of leafy vegetables is potent source of biological antioxidants. In addition, juice of raphanus, ajwain and sowa leaves possess capacity to mitigate starch induced postprandial glycemic burden and amaranthus leaves' juice can reduce glucose induced postprandial glycemic excursion.
  5 3,042 18
Identification of medical plants of 24 Ardisia species from China using the matK genetic marker
Yimei Liu, Ke Wang, Zhen Liu, Kun Luo, Shilin Chen, Keli Chen
October-December 2013, 9(36):331-337
DOI:10.4103/0973-1296.117829  PMID:24124285
Background: Ardisia is a group of famous herbs in China, which has been used as medical plants for more than 900 years. However, the species from the genus are so analogous that it is difficult to discriminate them just by morphological characteristics. DNA barcoding is a new technique that uses a short and standard fragment of DNA sequences to identify species. Objective: Choose a suitable DNA marker to authenticate Ardisia species. Materials and Methods: Four markers (psbA-trnH, internal transcribed spacer 2 [ITS2], rbcL, matK) were tested on 54 samples of 24 species from genus Ardisia. The success rates of polymerase chain reaction amplification and sequencing, differential intra- and inter-specific divergences, DNA barcoding gap and identification efficiency were used to evaluate the discrimination ability. Results : The results indicate that matK has the highest interspecific divergence and significant differences between inter- and intra-specific divergences, whereas psbA-trnH, ITS2 and rbcL have much lower divergence values. Matk possessed the highest species identification efficiency at 98.1% by basic local alignment search tool 1 [BLAST1], method and 91.7% by the nearest distance method. Conclusion: The matK region is a promising DNA barcode for the genus Ardisia.
  4 2,227 19
Determination of oleanolic acid and ursolic acid in Chinese medicinal plants using HPLC with PAH polymeric C18
Yong Zhang, Kunpeng Xue, Eugene Yuexing Zhao, Yin Li, Lixin Yao, Xiaoyan Yang, Xiaomei Xie
October 2013, 9(36):19-24
DOI:10.4103/0973-1296.117853  PMID:24143040
Background: The RP-HPLC resolution of two triterpenic acid isomers was unstable. Objective: To separate the oleanolic acid (OA) and ursolic acid (UA) simply within RP-HPLC. Materials and Methods: The separation ability of five stationary phases was studied with the retention effect of their carbon loads. Also the resolution effects of mobile phase composition and different column temperatures were systematically investigated by using Drylab; (Rheodyne LLC.) after evaluating chromatograms automatically. Results: The best available resolution of two bioactive isomers was achieved (r = 3.4) via using PAH (polycyclic aromatic hydrocarbons) polymeric C18 bonded phase column. The chromatographic system was applied to the quantification in ten Chinese medicinal plants and the validation was carried out and the precision (RSD ≤1.34%), the linearity (r ≥0.9998) and the recovery (range from 92.1% to 102.6%) were acceptable. Conclusion: It is clear that the method was simple, rapid and reliable for the quantification of two compounds in new HPLC method within PAH polymeric C18.
  4 3,120 27
Clinical Evaluation of Commiphora Mukul, a Botanical resin, in the Management of Hemorrhoids: A randomized controlled trial
Mahdi Yousefi, Mohammad Reza Vaez Mahdavi, Seyed Mousalreza Hosseini, Abdollah Bahrami, Ali Davati, Mohammad Kamalinejad, Sograt Faghihzadeh
October-December 2013, 9(36):350-356
DOI:10.4103/0973-1296.117832  PMID:24124288
Background : Hemorrhoids complaint is one of the most common problems in most society, especially in Asian countries. Current drug treatment protocols cannot cure the disease, and they are palliative. According to Persian traditional medicine, Commiphora Mukul (CM) resin is a medication choice. Aim : This randomized study was undertaken to evaluate the efficacy and safety of crude CM resin compared to a combination of lactolose and anti-hemorrhoid (LandA) in patients with uncomplicated hemorrhoids grade 1 and 2. Materials and Methods : This trial was carried out on 99 patients with hemorrhoids, in Ghaem and Imam Reaza Hospitals of the Mashhad University of Medical Sciences, Iran. They randomly received CM 3 g/d for 4 weeks (as study group) or LandA (Lactolose syrup in laxative dose for 1 month and anti-hemorrhoid suppository daily for 10 days) as control group. Subjective and objectives variables including painful defecation, flatulence, constipation, gastro-esophageal reflux (GER), dyspepsia, proctorrhagia, anal protrusion, and colonoscopic grading were assessed before, immediately after, and 4 weeks after the treatment period. An intent-to-treat analysis was used. Safety was assessed with evaluation of clinical adverse effects by common toxicity criteria version 4.0. Forty-nine patients were assigned randomly to receive LandA and 50 to receive CM. After 4 weeks, flatulence, dyspepsia, GER, and colonoscopic grading scores significantly decreased in study group, whereas in control group constipation, painful defecation, and proctorrhagia showed better but not significant improvement. After 4-weak follow-up, the rate of constipation, and proctorrhagia also showed significantly improvement in study group. Constipation and proctorrhagia in control group recurred significantly in 4-week follow-up than after the treatment, whereas this recurrence in test group was not seen. Conclusion : CM was more effective than LandA in 4-week treatment of patients with uncomplicated hemorrhoids grade 1 and 2.
  3 1,865 22
Diverse role of fast growing rhizobia in growth promotion and enhancement of psoralen content in Psoralea corylifolia L
Chandra Prabha, DK Maheshwari, Vivek K Bajpai
October 2013, 9(36):57-65
DOI:10.4103/0973-1296.117870  PMID:24143046
Background: Psoralea corylifolia (Bakuchi), a weed, which possesses a highly potent and medicinally important compound psoralen. P. corylifolia has been widely exploited since ages for its biological potential. Materials and Methods: Fifteen root nodulating bacteria as pure culture collection (PCC) were isolated from P. corylifolia in India. Further, these strains were evaluated for their effect on the psoralen content in P. corylifolia. High performance liquid chromatography (HPLC) method was used for the estimation of psoralen in P. corylifolia seed extracts. The effectiveness of these rhizobial strains was assessed on the basis of screening of various plant growth promoting attributes. Results: The 16S ribosomal RNA sequencing analysis revealed the identity of two most effective rhizobial isolates PCC2 and PCC7 as Rhizobium leguminosarum and Sinorhizobium meliloti, respectively. The R. leguminosarum PCC2 (JN546144) and Ensifer meliloti PCC7 (JN546145) strains showed solubilization of insoluble inorganic phosphate, secreted indole acetic acid (IAA), produced siderophore, showed ACC deaminase activity, and were positive for nodulation and nitrogen fixing genes. Seeds of P. corylifolia were bacterized with combination of R. leguminosarum PCC2 and Ensifer meliloti PCC7 along with their individual application that resulted in enhancement of various early vegetative and late reproduction parameters of plants in two consecutive field trials in the year 2009 and 2010. The psoralen content in the seeds of P. corylifolia was observed to be increased in the field trials where the combination of rhizobial strains PCC2 and PCC7 was used (2.79%) compared to control (1.91%). Conclusion: These findings indicate that rhizobial strains PCC2 and PCC7 showing good plant growth promoting attributes can be effective for increasing the psoralen content in the seeds of P. corylifolia to a certain level.
  3 2,162 16
Formulation development, optimization and evaluation of aloe vera gel for wound healing
Abdul Wadood Khan, Sabna Kotta, Shahid Hussain Ansari, Rakesh Kumar Sharma, Amit Kumar, Javed Ali
October 2013, 9(36):6-10
DOI:10.4103/0973-1296.117849  PMID:24143047
Purpose: To formulate and optimize a herbal gel of Aloe vera extract containing Carbopol 934 as gelling agent and to investigate the effects of topical application of Carbopol 934 gel containing Aloe vera extract on the healing of skin wounds surgically induced in Wistar rats. Materials and Methods: Different concentrations of viscosity enhancer Carbopol 934 were tried and finally gel that showed good spreadability and consistency was selected for wound healing property of herbal gel of Aloe vera. Excision wound model was used for the study. Results: The optimized gel was evaluated for different physicochemical properties and wound healing property. Differences in wound healing were observed between the various treatments when compared to the control group. Tissue hyperplasia was lower in the control group compared to the other treated groups. In animals group treated with gel, 80.14% healing was observed up to 14 th day. While in untreated group I (control) animals showed 52.68% healing of wounds on 14 th day. On the other hand, control group animals also showed inflammation and pus formation up to 5 th day of study, while treated animals did not showed any observable inflammation and pus formation. Conclusion: Results shows prepared gel has promising effect on the wound healing process.
  3 5,892 42
In vitro and in vivo studies of an aqueous extract of Matricaria recutita (German chamomile) on the radiolabeling of blood constituents, on the morphology of red blood cells and on the biodistribution of the radiopharmaceutical sodium pertechnetate
Angélica B Garcia-Pinto, Sebastião D Santos-Filho, Jorge J Carvalho, Mário J.S Pereira, Adenilson S Fonseca, Mário Bernardo-Filho
October 2013, 9(36):49-56
DOI:10.4103/0973-1296.117867  PMID:24143045
Background: Natural products might alter the labeling of blood constituents with technetium-99m ( 99m Tc) and these results may be correlated with modifications of the shape of the red blood cells (RBC). The biodistribution of radiopharmaceuticals can be also altered. Objective: This investigation aimed to determine biological effects of an aqueous extract of chamomile (CE). Materials and Methods: To study the effect of the CE on the labeling of blood constituents with 99m Tc, in vitro and in vivo assays were performed. The effect of the CE on the morphology of RBC was observed under light microscope. The images were acquired, processed, and the perimeter/area ratio of the RBC determined. To analyze the effect of the CE on biodistribution of the sodium pertechnetate (Na 99m TcO 4 ) in Wistar rats, these animals were treated or not with a CE. Na 99m TcO 4 was injected, the rats were sacrificed, the organs were removed, weighted and percentage of radioactivity/gram calculated. Result: In the in vitro experiment, the radioactivity on blood cells compartment and on insoluble fractions of plasma was diminished. The shape and the perimeter/area ratio of the RBC were altered in in vitro assays. An increase of the percentage of radioactivity of Na 99m TcO 4 was observed in stomach after in vivo treatment. Conclusion: These results could be due to substances of the CE or by the products of the metabolism of this extract in the animal organism. These findings are examples of drug interaction with a radiopharmaceutical, which could lead to misdiagnosis in clinical practice with unexpected consequences.
  2 2,110 13
The ginsenosides and carbohydrate profiles of ginseng cultivated under mountainous forest
Jian-kui Zhang, Rui Gao, De-qiang Dou, Ting-guo Kang
October 2013, 9(36):38-43
DOI:10.4103/0973-1296.117862  PMID:24143043
Background: Ginseng cultivated under mountainous forest, called "Lin-Xia-Shan-Shen" (LXSS) in China's Pharmacopoeia. In recent years, it has been quickly propelled to plant at a large scale. Objective: To study the profiles of ginsenosides and carbohydrate profiles of LXSS. Materials and Methods: The contents of ginsenosides and carbohydrates, such as soluble sugar, polysaccharide, pectin, and starch in LXSS, were determined. All the above components were profiled, and the correlations between them were analyzed. Results: The results indicated that the contents of total ginsenoside, protopanaxadiol, protopanaxatriol, Rg 1 , Re, Rb 1 , Rc, Rb 2 , Rd, starch, and pectin were negatively correlated with the growing years within 17 years. Among them, the content of starch was positively correlated with that of pectin. The total ginsenosides was positively correlated with starch and pectin, which cannot be found in garden ginseng, maybe resulting of fertilizer and other manual intervention in process of cultivation of garden ginseng. Discussion and Conclusions: The accumulation of ginsenosides and carbohydrate, especially starch and pectin, was different in garden ginseng and LXSS. This research may provide the scientific basis for germplasm evaluation, the cultivation and utilization of ginseng cultivated under mountainous forest.
  2 2,742 15
Effect of NR-Salacia on post-prandial hyperglycemia: A randomized double blind, placebo-controlled, crossover study in healthy volunteers
Pravina Koteshwar, Kadur Ramamurthy Raveendra, Joseph Joshua Allan, Krishnagouda Shankargouda Goudar, Kudiganti Venkateshwarlu, Amit Agarwal
October-December 2013, 9(36):344-349
DOI:10.4103/0973-1296.117831  PMID:24124287
Background: Salacia chinensis (S. chinensis) is widely distributed in India and Sri Lanka. Most of the species of genus Salacia are known to have effects on blood glucose levels; however, the effects of S. chinensis on glucose levels are seldom reported. Objective: To evaluate the oral hypoglycemic activity of NR- Salacia (1000 mg extract of S. chinensis) in healthy adults. Materials and Methods: Randomized, double-blind, placebo-controlled, cross-over study was conducted in healthy volunteers. Single dose of NR-Salacia (1000 mg extract of Salacia chinensis) and placebo were administered before carbohydrate-rich diet. A 6-point plasma glucose profile was performed at different time intervals up to 180 min. Results: NR-Salacia treatment significantly lowered plasma glucose level at 90 min, and the percentage reduction in glucose concentration was found to be 13.32 as compared to placebo group. A 33.85% decrease in the plasma glucose positive incremental area under curve (AUC) (0 to 180 min) was observed in comparison to placebo. No adverse events were recorded throughout the study period, except for some mild cases of abdominal discomforts like cramping and distention, vomiting, and headache in both placebo and NR-Salacia-treated groups. Conclusion: The study findings revealed that NR-Salacia lowered the post-prandial plasma glucose levels after a carbohydrate-rich meal and can be used as an oral hypoglycemic agent.
  2 2,013 16
Inhibition of wheat bran and it's active compoments on α-glucosidase in vitro
Jie Tu, Jun Chen, Shuyun Zhu, Chunxiao Zhang, Hua Chen, Youbing Liu
October-December 2013, 9(36):309-314
DOI:10.4103/0973-1296.117826  PMID:24124282
Background: Wheat bran is a traditional Chinese medicine; however, it is mostly used as feedstuff in China. Wheat bran is widely accepted as an important ingredient in many low-glycemic index foods in modern western societies; however, its glycemic control mechanism is unknown. Objective: To determine potent α-glucosidase inhibitory compounds from wheat bran and to identify the inhibition on α-glucosidase . Materials and Methods: Ethanolic extract of wheat bran was prepared to evaluate the inhibitory activity on α-glucosidase, then fractionation of the extract was guided by in vitro enzyme-inhibition assay, and the potent α-glucosidase inhibitory compounds were identified by high performance liquid chromatography and atmospheric pressure chemical ionization-mass spectrometry; finally the enzyme inhibition process was studied using the Michaelis-Menton and the Lineweaver-Burk equations. Results: Both baker's yeast and rat intestinal enzymes were mostly inhibited (87.9% and 66.8% inhibition, respectively) at concentration 0.6 mg/mL of the ethanolic extract of wheat bran. The petroleum ether fraction in the ethanolic extract of wheat bran showed significant activity against rat intestinal α-glucosidase, and revealed a dose-dependent effect. The inhibition was 76.57% at 0.3 mg/mL and 100% at 0.6 mg/mL. The active fraction 13 of petroleum ether fraction was identified as alkylresorcinols (ARs). ARs showed strong inhibition towards α-glucosidase and its IC 50 value was found to be 37.58 ΅g/mL. The enzyme kinetic studies showed that, in the presence of ARs, the Michaelis-Menton constant (Km ) remains constant whereas the maximal velocity (Vmax ) decreases, revealing a non-competitive type of inhibition. Conclusion: The therapeutic potentiality of ARs in the management of the postprandial hyperglycemia will proliferate the utilization of wheat bran in controlling type 2 diabetes.
  2 1,650 18
Ultra-performance liquid chromatography fingerprinting for quality control of Phragmitis rhizoma (Lugen) produced in Baiyangdian
Hong Li, Yu-Mei Gao, Jing Zhang, Lin Wang, Xiao-Xin Wang
October-December 2013, 9(36):285-289
DOI:10.4103/0973-1296.117810  PMID:24124278
Objective: To establish an ultra-performance liquid chromatography (UPLC) fingerprinting method for quality control of Phragmitis rhizoma from Baiyangdian. Materials and Methods: Ultrasonic extraction with 70% methanol was performed on 10 samples of P. rhizoma collected from 10 different villages in Baiyangdian. The sample solutions were analyzed by Waters UPLC equipped with the ACQUITY UPLC BEH C 18 column and photodiode array (PDA) detector, and gradient eluted with acetonitrile/water as the mobile phase. The flow rate was set to 0.1 mL/min; the column temperature was set to 25°C; and the detection wavelength was set to 285 nm. Results: The chromatograms of the 10 samples showed 27 common peaks, of which one was identified as the ferulic acid standard. The similarity indexes were all above 0.82. Hierarchical cluster analysis showed that the constituents and their quantities differed according to the diameter of the original plant, which is related to its age. Conclusion: The UPLC fingerprinting method had the advantages of being fast, accurate, and highly efficient; this indicated that it can be used for quality control of P. rhizoma produced in Baiyangdian. Also, the relation between the quality and diameter/age of the plant needs to be further investigated.
  1 1,878 18
β-cyclodextrin assistant flavonoid glycosides enzymatic hydrolysis
Xin Jin, Zhen-hai Zhang, E Sun, Xiao-Bin Jia
October 2013, 9(36):11-18
DOI:10.4103/0973-1296.117851  PMID:24143039
Background: The content of icaritin and genistein in herba is very low, preparation with relatively large quantities is an important issue for extensive pharmacological studies. Objective: This study focuses on preparing and enzymic hydrolysis of flavonoid glycosides /β-cyclodextrin inclusion complex to increase the hydrolysis rate. Materials and Methods: The physical property of newly prepared inclusion complex was tested by differential scanning calorimetry (DSC). The conditions of enzymatic hydrolysis were optimized for the bioconversion of flavonoid glycosides /β-cyclodextrin inclusion complex by mono-factor experimental design. The experiments are using the icariin and genistein as the model drugs. Results: The solubility of icariin and genistein were increased almost 17 times from 29.2 μg/ml to 513.5 μg/ml at 60˚ C and 28 times from 7.78 μg/ml to 221.46 μg/ml at 50˚ C, respectively, demonstrating that the inclusion complex could significantly increase the solubility of flavonoid glycosides. Under the optimal conditions, the reaction time of icariin and genistin decreased by 68% and 145%, when compared with that without β-CD inclusion. By using this enzymatic condition, 473 mg icaritin (with the purity of 99.34%) and 567 mg genistein(with the purity of 99.46%), which was finally determined by melt point, ESI-MS, UV, IR, 1 H NMR and 13 C NMR, was obtained eventually by transforming the inclusion complex(contains 1.0 g substrates). Conclusion: This study can clearly indicate a new attempt to improve the speed of enzyme-hydrolysis of poorly water-soluble flavonoid glycosides and find a more superior condition which is used to prepare icaritin and genistein.
  1 2,590 18
The stability investigation of compound Danshen injection (a traditional medicine) with a new high-performance liquid chromatography method
Yanqing Zhang, Fengqin Bao, Zhixia Zhao, Xiuyu Sun, Wuqin Qi, Junbo Xie
October-December 2013, 9(36):338-343
DOI:10.4103/0973-1296.117830  PMID:24124286
Background: Compound Danshen injection (CDSI, a traditional medicine) is an effective drug for the treatment of cardiovascular and cerebrovascular diseases. However, the research about its stability is absent. Objective: A new high-performance liquid chromatography method was developed to assay its main effective constituents, i.e., propanoid acid (PA), protocatechuic aldehyde (PHA), salvianolic acid B (SAB), salvianolic acid A (SAA), and rosmarinic acid (RA). Through the newly found method, the stability of CDSI was to be investigated. Materials and Methods: The analysis was performed by a reverse-phase gradient elution using an aqueous mobile phase (containing 0.1% acetic acid) modified by acetonitrile, and detection was made simultaneously at 280 nm and 325 nm. The method was validated for accuracy, precision and limits of detection. The effects of some environmental storage conditions (light and temperature) on the stability of CDSI were investigated. Results: This method is precise, simple, and convenient. The result showed that illumination and temperature had an obvious effect on CDSI's stability. SAA is the most unstable one among the five components. In the condition of common light, it decomposed rapidly to almost 50% after only 4 h, and 100% after 8 h. PA, RA, and PHA might come from Danshen, was also the transformed products from other components in store process. Conclusion: The result indicated that the main active constituents in CDSI suffered from the illumination and temperature greatly. CDSI should be stored at low temperature and kept away from light.
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EDITORIAL
Pharmacognosy Magazine's new and improved Impact Factor - 1.525
S Nandan
October-December 2013, 9(36):283-284
DOI:10.4103/0973-1296.117807  PMID:24124277
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ORIGINAL ARTICLES
Influence of the mixture of Epimedii Herba and Ginkgo Folium extracts on the coronary flow of isolated hearts in rats
DongFang Zhang, ChangJi Yuan, Zheng Zhu, Xin Jin, LiHong Li
October-December 2013, 9(36):290-293
DOI:10.4103/0973-1296.117816  PMID:24124279
Background: In clinical practice, Epimedii Herba and Ginkgo Folium preparations are widely used in treatment of diseases such as coronary heart disease (angina) in China . However, there are no studies on the two-drug combination. Objective: To explore the effect of the mixture of the Epimedii Herba extract (EE) and Ginkgo Folium extract (GE) on coronary flow of isolated hearts in rats. Materials and Methods : EE and GE were prepared by reflux in alcohol, and processed with HPD-100 macro-reticular resins; icariin from EE and total bilobalides from GE were determined by high performance liquid chromatography (HPLC). Fifty male Sprague-Dawley (SD) mice were subdivided into five groups (10 rats each): Normal control group (NC), EE - 10 mg group, GE - 10 mg group, EE - 5 mg + GE - 5 mg group, and EE - 10 mg + GE - 10 mg group. Isolated hearts uniform pressure perfusion was proceeded with Langendorff system. Results: The content of icariin in EE was 20.8%. The total content including four kinds of bilobalides (ginkolide A-C and bilobalide) in GE was 8.6%. The coronary flow in the NC group remained stable before and after treatment, and the coronoray flow in the EE, GE, EE + GE groups was increased and the relative magnitude of heightening was 25.0-33.3%, and the coronary flow in EE + GE was significantly different from that in the single EE or GE group. Conclusion: EE or GE itself can heighten coronary flow of isolated hearts in rats. The activity of the mixture including EE and GE is better than that of single EE or GE, and the activity becomes larger when the dosage is doubled, and is related with dosage.
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Tissue culture of Sophora tonkinensis Gapnep. and its quality evaluation
Wei Kun-Hua, Li Lin-Xuan, Huang Yong-Cai, Wang Mei-Ying, Li Cui, Miao Jian-Hua
October-December 2013, 9(36):323-330
DOI:10.4103/0973-1296.117828  PMID:24124284
Background: Sophora tonkinensis Gapnep. is an important rare medicinal plant in China. There were only a few papers on the rapid propagation of S. tonkinensis through in vitro tissue culture, and still no report focuses on the quality analysis of in vitro tissue culture plantlets. Materials and Methods: The different concentrations of 6-benzylaminopurine (BAP), kinetin (KT), and indole-3-acetic acid (IAA) were used to establish and screen the optimal rapid propagation technology of S. tonkinensis by orthogonal test; the different concentrations of a-naphthalene acetic acid (NAA), indole-3-butyric acid (IBA), and ABT rooting power (ABT) were used to screen the optimal rooting technology. For quality evaluation of tissue culture plants, three different sites were chose to finish planting experiment. The leaf characteristics, radix ex rhizoma yield, and contents of matrine and oxymatrine were evaluated, respectively, to provide evidence of high yield and good qualities of tissue culture plants. Results: A large number of buds could be induced directly from epicotyl and hypocotyl explants on the Murashige and Skoog (MS) medium supplemented with 1.5 mg/l BAP, 0.5 mg/l IAA, and 0.5 mg/l KT; the best root induction medium was solid MS medium at half the macronutrient concentration supplemented with 1.0 mg/l NAA, 0.4 mg/l IBA, and 0.1 mg/l ABT. The rooting rate was 98%. All tissue culture plants showed normal leaf characteristics. Tissue culture plants from two sites possessed higher radix ex rhizoma yield and overall productivity of matrine and oxymatrine than those of seed plants. Conclusion: Tissue culture is a rapid, effective, and convenient propagation method for S. tonkinensis, and the quality of S. tonkinensis tissue culture plants meets the requirement of quality standard of China Pharmacopoeia (edition 2010), the crude drug from S. tonkinensis tissue culture plants will be suitable for substituting the crude drug from seed plants.
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Benzophenones from Hypericum elegans with antioxidant and acetylcholinesterase inhibitory potential
Dimitrina Zheleva-Dimitrova, Paraskev Nedialkov, Georgi Momekov
October 2013, 9(36):1-5
DOI:10.4103/0973-1296.117846  PMID:24143038
Background: Hypericum elegans is used in Bulgarian folk medicine for treatment of wounds, depression, gastrointestinal and bacterial diseases. Objective: Recently, new natural benzophenones: Elegaphenone and O-glycosides: Hypericophenonoside, Neoannulatophenonoside and Elegaphenonoside as well as already known 7-Epiclusianone were isolated from the titled species. The aim of the present study was to evaluate the antioxidant and acetyl cholinesterase inhibitory potential of the isolated compounds. Materials and Methods: 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) di-ammonium salt (ABTS) free radicals, ferric reducing antioxidant power (FRAP) assay as well as inhibition of lipid peroxidation in linoleic acid system were used for determination of antioxidant activity. Modified Ellman's colorimetric method was carried out to assess the acetyl cholinesterase inhibition potential. Hyperoside and Galantamine hydrobromide were used as positive controls. Results: Hypericophenonoside was found to possess the strongest DPPH radical scavenging activity (IC 50 = 181.85 ± 6.82 μM), while Neoannulatophenonoside showed the highest ABTS (IC 50 = 0.25 ± 0.005 μM) and lipid peroxidation inhibitor activity. FRAP activity was demonstrated only by prenylated aglycones - Elegaphenone [942.16 ± 4.03 μM Trolox Equivalent (TE)] and 7-Epiclusianone (642.95 ± 3.95 μM TE) and was stronger compared to the control Hyperoside (421.75 ± 9.29 μM TE). Elegaphenone and 7-Epiclusianone were found to possess moderate acetyl cholinesterase inhibitory potential with IC 50 values of 192.19 ± 3.54 μM and 142.97 ± 4.62 μM, respectively. Conclusion: The results obtained revealed that H. elegans is a potential natural source of bioactive compounds and benzophenones could be useful in therapy of free radical pathologies and neurodegenerative disorders.
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Bioassay- and liquid chromatography/mass spectrometry-guided acetylcholinesterase inhibitors from Picriafel-terrae
Lu Wen, Qiqiu Wei, Gang Chen, Fan Liu, Shichang Zhang, Tinghuo You
October 2013, 9(36):25-31
DOI:10.4103/0973-1296.117857  PMID:24143041
Background: Picria fel-terrae is a traditional Chinese medicine. Materials and Methods: A new approach to the search for acetylcholinesterase (AChE) inhibitors from Picria fel-terrae is presented. Results: Bioassay- and LC-MS-guided fractionation of the ethyl acetate extract was from traditional Chinese medicine P.fel-terrae. Following primary extraction, the ethyl acetate extracts fraction of P.fel-terrae showed strong AChE inhibitory activities. So the sample was separated using highperformance liquid chromatography (HPLC). The effluent was split towards two identical 96-well fraction collectors, and the presence of the biologically interesting portion and chromatographic fractions could be readily detected by analyzing selected ion chromatograms through an electrophoresis-electrospray ionization mass spectrometry (ESIMS) system for accurate mass measurement. One 96-well plate was used for a bioassay (AChE-inhibitory assay) and detected the bioactivity and position of the relevant peak in the chromatogram. The positive well in the second 96-well plate was used for identification by LC-(+) ESIMS. Conclusion: As abovementioned, the AChE inhibitory constituents from P.fel-terrae by LC-bioassay-ESIMS were rapid identified. Liquid chromatography/ mass spectrometry (LC-MS) screening detected the presence of six active compounds, identified as picfeltarraenin IA (1), picfeltarraenin IB (2), picfeltarraenin IV (3), picfeltarraenin X (4), picfeltarraenin XI (5), and one unknown compound. The structures were further determined by 13C NMR. The six compounds expressed stronger AChE inhibition than the known AChE inhibitorTacrine. Above all, the value of this LC-bioassay-ESIMS methodology is highlighted by the finding and structure elucidation of the active constituents from many other structural families of natural products.
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