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   2014| August  | Volume 10 | Issue 39  
    Online since August 30, 2014

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Efficacy of ellagic acid and sildenafil in diabetes-induced sexual dysfunction
Sumanta Kumar Goswami, Manikanta Vishwanath, Suma Kallahalli Gangadarappa, Rema Razdan, Mohammed Naseeruddin Inamdar
August 2014, 10(39):581-587
DOI:10.4103/0973-1296.139790  PMID:25298678
Background: Diabetes induced sexual dysfunction is a leading cause of male sexual disorder and an early indicator of cardiovascular complication. Reactive oxygen species generated in body during diabetes is a main causative factor for erectile dysfunction, a sexual dysfunction. Adjuvant antioxidant therapy along with phosphodiesterases type 5 enzyme inhibitor (PDE5i) is more effective than PDE5i alone. Objective: The aim of the study was to investigate efficacy of ellagic acid a known antioxidant and sildenafil in diabetes induced erectile dysfunction. Materials and Methods: Type 1 diabetes was induced in male rats and rats were treated with ellagic acid (50 mg/kg, p.o.) and a combination of ellagic acid (50 mg/kg, p.o.) and sildenafil (5 mg/kg, p.o.), a PDE5i for 28 days. Sexual function was observed in diabetic rat and compared with those of treatment group and normal rats. Effect of ellagic acid was studied on advanced glycation end products (AGE) and isolated rat corpus cavernosum in vitro. Results: Sexual function of diabetic rats was found to be reduced and ellegic acid treatment could preserve sexual function of diabetic rats to some extent. Ellagic acid + sildenafil treatment was more efficient in management of diabetes induced sexual dysfunction. Ellagic acid inhibited (AGE) in vitro implying its role in reducing oxidative stress in diabetes. The polyphenol could not increase sexual function in normal rats and relax isolated rat corpus cavernosum smooth muscle significantly. Conclusion: The study proves usefulness of adjuvant antioxidant therapy in the management of erectile dysfunction in diabetes.
  19 4,707 151
Content determination of the flavonoids in the different parts and different species of Abelmoschus esculentus L. by reversed phase-high performance liquid chromatograph and colorimetric method
Yin Lin, Min-feng Lu, Hai-bing Liao, Yu-xian Li, Wei Han, Ke Yuan
July-September 2014, 10(39):278-284
DOI:10.4103/0973-1296.137368  PMID:25210315
Background: This research will establish the ultraviolet colorimetric method to determine the total flavonoid content in different species and different parts of Abelmoschus esculentus L. Materials and Methods: We establish the reversed-phase high-performance liquid chromatograph (RP-HPLC) method to determine the content of the three flavonoid glycosides in different species and different parts of the A. esculentus. Adopt the NaNO 2 -Al (NO 3 ) 3 -NaOH colorimetric method to determine the total flavonoid content; at the same time, adopt the RP-HPLC method to determine the contents of the three flavonoid glycosides. Using the methods of ultraviolet colorimetry and RP-HPLC, we determined and analyzed the total flavonoid content and the content of the three flavonoid glycosides in different species and different parts of A. esculentus. Results: There are great distribution differences of the total flavonoids and the three flavonoid glycosides in different species and parts of A. esculentus. Among them, the content of the effective constituents in the flower is relatively high, next is in the fruit. In the different species of A. esculentus, the content of the flavonoids of finger relatively high. The HPLC method established in this research is simple and convenient and its results are accurate and reliable. In addition, it has a very good repeatability. Conclusion: The results provided the reference data for the medicinal use of A. esculentus and it can be used in quality analyzing of its effective constituents.
  16 3,641 157
LC-MS/MS determination and pharmacokinetics study of puerarin and daidzein in rat plasma after oral administration of Gegenqinlian decoction and Radix Puerariae extract
Yifan Zhang, Jin Yuan, Ying Wang, Yue Wang, Rui An, Xinhong Wang
July-September 2014, 10(39):241-248
DOI:10.4103/0973-1296.137363  PMID:25210310
Background: Gegenqinlian decoction (GQD) is a famous traditional medicine recipe. It is composed of four herbs including Radix Puerariae (GG), Radix Scutellariae (HQ), Rhizoma Coptidis (HL) and Radix Glycyrrhizae (GC), which is widely used for treating gastro-intestinal disorders in the clinical practice of Traditional Chinese medicine (TCM). The aim of this study was to compare the pharmacokinetics of puerarin and daidzein in rats following oral administration of Gegenqinlian Decoction and Radix puerariae extract. Thus, a sensitive and selective liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was developed and validated for simultaneous determination of puerarin and daidzein in rat plasma following oral administration of Gegenqinlian Decoction and Radix Puerariae extract. Materials and Methods: Chromatographic separation was performed on a Shiseido CAPCELL PAK C 18 analytical column (100 mm × 2.0 mm i.d., 5 μm) by linear gradient elution, with water (0.1% formic acid)-acetonitrile (0.1% formic acid) as mobile phase. Detection was carried out by multiple reaction monitoring (MRM) mode using electrospray ionization in the positive ion mode. Results: The calibration curves were linear over a range of 7.80-1560 ng/mL for puerarin and 6.30-1260 ng/mL for daidzein. The intra- and inter-day precision values were less than 13.6% and their average recoveries was in the range of 77.8% and 88.6% for puerarin and was between 76.3 and 86.8% for daidzein, respectively. Conclusion: The validated method was applied to the comparative pharmacokinetic studies of puerarin and daidzein after oral administration of Gegenqinlian Decoction and Radix Puerariae extract. The pharmacokinetic parameters showed that puerarin and daidzein from Gegenqinlian Decoction were absorbed more effectively with slower elimination in rat plasma than that from Radix Puerariae extract. These results revealed that as far as the Radix Puerariae extract was concerned, it is very valuable to be used as a clinical directions of Gegenqinlian Decoction.
  13 4,102 184
Identification of natural inhibitors against angiotensin I converting enzyme for cardiac safety using induced fit docking and MM-GBSA studies
Balakrishnan Vijayakumar, Subramani Parasuraman, Ramasamy Raveendran, Devadasan Velmurugan
August 2014, 10(39):639-644
DOI:10.4103/0973-1296.139809  PMID:25298685
Background: Cleistanthins A and B are isolated compounds from the leaves of Cleistanthus collinus Roxb (Euphorbiaceae). This plant is poisonous in nature which causes cardiovascular abnormalities such as hypotension, nonspecific ST-T changes and QTc prolongation. The biological activity predictions spectra of the compounds show the presence of antihypertensive, diuretic and antitumor activities. Objective: Objective of the present study was to determine the in silico molecular interaction of cleistanthins A and B with Angiotensin I- Converting Enzyme (ACE-I) using Induced Fit Docking (IFD) protocols. Materials and Methods: All the molecular modeling calculations like IFD docking, binding free energy calculation and ADME/Tox were carried out using Glide software (Schrφdinger LLC 2009, USA) in CentOS EL-5 workstation. Results: The IFD complexes showed favorable docking score, glide energy, glide emodel, hydrogen bond and hydrophobic interactions between the active site residues of ACE-I and the compounds. Binding free energy was calculated for the IFD complexes using Prime MM-GBSA method. The conformational changes induced by the inhibitor at the active site of ACE-I were observed based on changes of the back bone Cα atoms and side-chain chi (x) angles. The various physicochemical properties were calculated for these compounds. Both cleistanthins A and B showed better docking score, glide energy and glide emodel when compared to captopril inhibitor. Conclusion: These compounds have successively satisfied all the in silico parameters and seem to be potent inhibitors of ACE-I and potential candidates for hypertension.
  13 3,040 147
Antidepressant-like effects of Chaihu-Shugan-San via SAPK/JNK signal transduction in rat models of depression
Yun-Hui Li, Chun-Hu Zhang, Juan Qiu, Su-E Wang, Sui-Yu Hu, Xi Huang, Ying Xie, Yang Wang, Tian-Li Cheng
July-September 2014, 10(39):271-277
DOI:10.4103/0973-1296.137367  PMID:25210314
Background: Chaihu-Shugan-San (CHSGS), a traditional Chinese medicinal herbal formula, registered in Jingyue Quanshu, has been indicated that oral administration of the extract from it can remit depressive disorder. C-Jun amino-terminal kinase (JNK/SAPK) signal transduction plays a key role in the apoptosis of nerve cells, be reported closely correlated with depression. This study was designed to investigate CHSGS antidepressant-like effects in rat models of depression and probe its possible mechanism. Materials and Methods: The classical experimental depression model chronic mild unpredictable stress (CMUS) was used to evaluate the antidepressant-like effects of CHSGS. The extracts were administered orally for 14 days, while the parallel positive control was given at the same time using fluoxetine hydrochloride. The expressions of JNK in the hippocampus were detected by real-time fluorescent quantitation PCR and Western blot assay. Results: Intragastric administration of CHSGS for 14 days caused a significant improvement of weight and locomotor activity in the open-field test. In addition, CHSGS treatment inhibited the expressions of JNK in the hippocampus tissue in CMUS rats. Conclusion: CHSGS could obviously improve the depressive state of the model rats and its mechanism may be correlated with regulating the expressions of JNK in the hippocampus.
  12 3,907 101
Anticancer activity of flavane gallates isolated from Plicosepalus curviflorus
Ghada Ahmed Fawzy, Areej Mohammad Al-Taweel, Shagufta Perveen
August 2014, 10(39):519-523
DOI:10.4103/0973-1296.139787  PMID:25298669
Background: Previous investigation of the methanol extract of Plicosepalus curviflorus leaves led to the isolation of two new flavane gallates (1, 2), together with other compounds including quercetin (3). The stems of P. curviflorus are used traditionally for the treatment of cancer in Yemen. Objective: The aim of this study was to evaluate the anticancer activity of the plant methanol extract as well as isolated compounds (1-3). Materials and Methods: The human cancer cell lines used were; MCF-7, HepG-2, HCT-116, Hep-2, HeLa and normal, Vero cell line using the Crystal Violet Staining method (CVS). Results: Quercetin (3) possessed the highest anticancer effect against all five cell lines (IC 50 ranging from 3.6 to 16.2 μg/ml). It was followed by 2S, 3R-3, 3′, 4′, 5, 7-pentahydroxyflavane-5-O-gallate (1) , with IC 50 ranging from 11.6 to 38.8 μg/ml. The weakest anticancer activity was given by 2S, 3R-3,3′,4′,5,5′,7-hexahydroxyflavane-3′,5-di-O-gallate (2) with IC 50 ranging from 39.8 to above 50 μg/ml, compared to vinblastine sulphate as reference drug. Colon, liver and breast cell lines seemed to be more sensitive to the tested compounds than the cervical and laryngeal cell lines. Concerning the cytotoxic effect on Vero cell line, the pentahydroxyflavane-5-O-gallate (1) showed the highest IC 50 ( 138.2 μg/ml), while quercetin exhibited the lowest IC 50 to Vero cells (30.5 μg/ml), compared to vinblastine sulphate as reference drug (IC 50: 39.7 μg/ml). Conclusion: The results suggest the possible use of compounds 1 and 3 as anticancer drugs especially against colon and liver cancers.
  11 3,279 102
Efficient production of flavonoids in Fagopyrum tataricum hairy root cultures with yeast polysaccharide elicitation and medium renewal process
Jiang-Lin Zhao, Liang Zou, Cai-Qiong Zhang, Yuan-Yuan Li, Lian-Xin Peng, Da-Bing Xiang, Gang Zhao
July-September 2014, 10(39):234-240
DOI:10.4103/0973-1296.137362  PMID:25210309
Background: Tartary buckwheat (Fagopyrum tataricum), an excellent edible and medicinal crop, has been widely used as a daily diet and traditional medicine for a long time. The major functional components of Fagopyrum tataricum have been demonstrated to be flavonoids (i.e. rutin and quercetin), which had notable andioxidant, antidiabetic, hypocholesterolemic and antitumor activities. Hairy root culture is a convenient and efficient plant tissue culture system for large scale production of bioactive metabolites. Objective: To enhance the functional flavonoids production in hairy root culture of F. tataricum. Materials and Methods: The elicitation treatment in combination with medium renewal strategy was applied for efficient promoting flavonoids production in F. tataricum hairy root cultures. Results: The exogenous yeast polysaccharide (YPS) elicitor notably stimulated the functional metabolites production in F. tataricum hairy root cultures, and the stimulation effect was concentration-dependent. Combination with the YPS elicitation (200 mg/L) and medium renewal process, the maximal flavonoids yield was enhanced to 47.13 mg/L, about 3.2-fold in comparison with the control culture of 14.88 mg/L. Moreover, this research also revealed the accumulation of these bioactive metabolites resulted from the stimulation of the phenylpropanoid pathway by YPS treatment. These results indicated that the F. tataricum hairy root culture could be an effective system for rutin and quercetin production.
  10 4,197 97
Anticancer activity of cissampelos pareira against dalton's lymphoma ascites bearing mice
B Samuel Thavamani, Molly Mathew, SP Dhanabal
July-September 2014, 10(39):200-206
DOI:10.4103/0973-1296.137356  PMID:25210304
Background: Cissampelos pareira (Menispermaceae) is used in folk Indian system of alternative medicine, for its analgesic, antipyretic, diuretic, antilithic, and emmenagogue properties. Objective: To evaluate Cissampelos pareira (C. pareira) for in vitro cytotoxicity and in vivo antitumor activity against Dalton's Lymphoma Ascites (DLA) cells in Swiss mice. Materials and Methods: Cissampelos pareira was successively extracted using different solvents. In vitro cytotoxicity was assessed by the MTT assay. An in vivo study was carried out in methanol extract. Twenty-four hours after intraperitoneal inoculation of the DLA cells in mice, the methanol extract of C. pariera (MECP) was administered at 200 and 400 mg/kg body weight for 14 consecutive days. On day 14, six mice were sacrificed and the rest were kept alive for assessment of increase in life-span. The antitumor effect was assessed by evaluating the packed cell volume, viable tumor cell count, increase in body weight, and increase in life-span. The hematological and serum biochemical parameters and anti-oxidant properties were assessed by estimating the superoxide dismutase (SOD), catalase (CAT), and lipid peroxidation. Results: Methanol Extract of Cissampelos pariera (MECP) showed a potent cytotoxic activity, with an IC 50 value of 95.5 μg/ml and a significant (P < 0.001) decrease in packed cell volume, viable cell count, and an increased lifespan (54 and 72%). The hematological and serum biochemical profiles were restored to normal levels in MECP-treated mice. The MECP-treated group significantly (P < 0.001) decreased SOD, lipid peroxidation, and CAT to normal. Conclusion: This study demonstrated that C. pariera exhibited significant in vitro and in vivo anti-tumor activities and that it was reasonably imputable to its increasing endogenous mechanism of antioxidant property.
  9 4,894 96
In vivo anti-fatigue activity of sufu with fortification of isoflavones
Yunxian Liu, Yun Zhou, Satoru Nirasawa, Eizo Tatsumi, Yongqiang Cheng, Lite Li
July-September 2014, 10(39):367-373
DOI:10.4103/0973-1296.137380  PMID:25210327
Background: Sufu is a traditional Chinese fermented soybean food. Isoflavones are abundant in soybean and products incorporated with isoflavones exert many health benefits. The aim of this study was to investigate anti-fatigue effect of sufu fortified with isoflavones. Materials and Methods: In vivo anti-fatigue activity of sufu with fortification of isoflavones (IF) was investigated in this study via exhaustive swimming test using ICR mice and determination of biochemical parameters. Factors relating to fatigue, including hepatic glycogen, blood lactic acid (BLA), blood urea nitrogen (BUN) were determined. The isoflavone composition in the IF sufu was also determined to explore the anti-fatigue activity of isoflavones. Results: During fermentation, isoflavone glucosides were converted into aglycones and both sufu with and without fortification of IF prolonged the exhaustive swimming time of ICR mice. Intake of sufu also increased the hepatic glycogen content, while it decreased the levels of both the blood lactic acid (BLA) and blood urea nitrogen (BUN) content. A dose-response relationship was observed in both exhaustive swimming and BLA clearance test, with medium dose (1%) fortification of IF revealing the highest activity. Conclusion: IF sufu could possess high anti-fatigue activity.
  9 3,184 104
Anti-melanogenic activity of the novel herbal medicine, MA128, through inhibition of tyrosinase activity mediated by the p38 mitogen-activated protein kinases and protein kinase signaling pathway in B16F10 cells
Aeyung Kim, Jin Yeul Ma
August 2014, 10(39):463-471
DOI:10.4103/0973-1296.139774  PMID:25298661
Background: Recently, our research group developed MA128, a novel herbal medicine, and demonstrated that MA128 is effective for the treatment of asthma and atopic dermatitis (AD). In particular, postinflammatory hyper-pigmentation in AD mice was improved with MA128 treatment. Thus, in this study, we determined the effect of MA128 on melanogenesis and its underlying mechanism in murine B16F10 melanoma cells. Materials and Methods: After treatment with MA128 at 100 and 250 μg/mL and/or alpha-melanocyte stimulating hormone (α-MSH) (1 μM), cellular melanin content and tyrosinase activity in B16F10 cells were measured. Using western blotting, expression levels of tyrosinase, tyrosinase-related protein-1 (TRP-1), TRP-2, microphthalmia-associated transcription factor (MITF), and activation of c-AMP-dependent protein kinase (PKA), c-AMP-related element binding protein (CREB) and mitogen-activated protein kinases (MAPKs) were examined. Results: MA128 significantly inhibited melanin synthesis and tyrosinase activity in a resting state as well as α-MSH-stimulating condition, and significantly decreased the expression of tyrosinase, TRP-1, TRP-2 and MITF. In addition, phosphorylation of PKA and CREB by α-MSH stimulation was efficiently blocked by MA128 pretreatment. Moreover, MA128 as an herbal mixture showed synergistic anti-melanogenic effects compared with each single constituent herb. Conclusion: MA128 showed anti-melanogenic activity through inhibition of tyrosinase activity mediated by p38 MAPK and PKA signaling pathways in B16F10 cells. These results suggest that MA128 may be useful as an herbal medicine for controlling hyper-pigmentation and as a skin-whitening agent.
  8 4,174 169
Qualitative and quantitative analysis of the major constituents in traditional Chinese medicine Danmu injection using LC-ESI-MS n and LC-DAD
Fen-Xia Zhu, Jing-Jing Wang, Xiu-Feng Li, E Sun, Xiao-Bin Jia
July-September 2014, 10(39):254-264
DOI:10.4103/0973-1296.137365  PMID:25210312
Background: Danmu injection, a traditional Chinese medicine (TCM) preparation made from Nauclea officinalis, has been commonly used for the treatment of cold, fever, swelling of throat in China. However, the chemical constituents in Danmu injection have not been clarified yet. Objective: a HPLC/DAD/ESI-MS n method was developed for qualitative and quantitative analysis of the components in Danmu injection. Materials and Methods: The chromatographic separation was performed on a Welch Material XB-C18 (4.6mm × 250mm, 5μm) using gradient elution with acetonitrile (A) and water containing 0.1% formic acid (B) as mobile phase at a flow rate of 1.0 ml/min. Results: Twenty-five compounds, including phenolic acid and phenol glycoside, iridous glycoside and glycoalkaloid were identified or tentatively deduced on the base of their retention behaviors, UV absorption, MS and MS n data with those elucidated references or literature. In addition, eleven compounds were simultaneously determined by HPLC-DAD, which was validated and successfully applied for determination of major components in Danmu injection. Conclusion: The results suggested that the established qualitative and quantitative method would be a powerful and reliable analytical tool for the characterization of multi-constituent in complex chemical system and quality control of Danmu injection.
  8 3,378 130
Chemical constituents and toxicological studies of leaves from Mimosa caesalpiniifolia Benth., a Brazilian honey plant
Nayana Bruna Nery Monção, Luciana Muratori Costa, Daniel Dias Rufino Arcanjo, Bruno Quirino Araújo, Maria do Carmo Gomes Lustosa, Klinger Antônio da França Rodrigues, Fernando Aécio de Amorim Carvalho, Amilton Paulo Raposo Costa, Antônia Maria das Graças Lopes Citó
August 2014, 10(39):456-462
DOI:10.4103/0973-1296.139773  PMID:25298660
Background: Mimosa caesalpiniifolia Benth. (Leguminosae) is widely found in the Brazilian Northeast region and markedly contributes to production of pollen and honey, being considered an important honey plant in this region. Objective: To investigate the chemical composition of the ethanol extract of leaves from M. caesalpiniifolia by GC-MS after derivatization (silylation), as well as to evaluate the in vitro and in vivo toxicological effects and androgenic activity in rats. Materials and Methods: The ethanol extract of leaves from Mimosa caesalpiniifolia was submitted to derivatization by silylation and analyzed by gas chromatography-mass spectrometry (GC-MS) to identification of chemical constituents. In vitro toxicological evaluation was performed by MTT assay in murine macrophages and by Artemia salina lethality assay, and the in vivo acute oral toxicity and androgenic evaluation in rats. Results: Totally, 32 components were detected: Phytol-TMS (11.66%), lactic acid-2TMS (9.16%), α-tocopherol-TMS (7.34%) and β-sitosterol-TMS (6.80%) were the major constituents. At the concentrations analyzed, the ethanol extract showed low cytotoxicity against brine shrimp (Artemia salina) and murine macrophages. In addition, the extract did not exhibit any toxicological effect or androgenic activity in rats. Conclusions: The derivatization by silylation allowed a rapid identification of chemical compounds from the M. caesalpiniifolia leaves extract. Besides, this species presents a good safety profile as observed in toxicological studies, and possess a great potential in the production of herbal medicines or as for food consumption.
  7 1,632 24
Effects of rhamnocitrin 4-β-D-galactopyranoside, isolated from Astragalus hamosus on toxicity models in vitro
Magdalena Kondeva-Burdina, Ilina Krasteva, Mitka Mitcheva
August 2014, 10(39):487-493
DOI:10.4103/0973-1296.139778  PMID:25298664
Background: Astragalus hamosus L. (Fabaceae) is used in herbal medicine as emollient, demulcent, phrodisiac, diuretic, laxative, and good for inflammation, ulcers, and leukoderma. It is useful in treating irritation of the mucous membranes, nervous affections, and catarrh. Objective: Rhamnocitrin 4-β-D-galactopyranoside (RGP), isolated from A. hamosus, was investigated for its possible protective effect on different models of toxicity in vitro on sub-cellular and cellular level. Materials and Methods: The effects of RGP were evaluated on isolated rat brain synaptosomes, prepared by Percoll reagent and on rat hepatocytes, isolated by two-stepped collagenase perfusion. Results: In synaptosomes, RGP had statistically significant protective effect, similar to those of silymarin, on 6-hydroxy (OH)-dopamine-induced oxidative stress. These results correlate with literature data about protective effects of kempferol and rhamnocitrin on oxidative damage in rat pheochromocytoma PC12 cells. In rat hepatocytes, we investigate the effect of RGP on two models of liver toxicity: Bendamustine and cyclophosphamide. In these models, the compound had statistically significant cytoprotective and antioxidant activity, similar to those of silymarin. Conclusion: According to these results, we can suggest that such cytoprotective effect of RGP might be due to an influence on bendamustine and cyclophosphamide metabolism in rat hepatocytes. In isolated rat hepatocytes, in combination with bendamustine and cyclophosphamide and in 6-OH-dopamine-induced oxidative stress in isolated rat synaptosomes, RGP, isolated from A. hamosus, was effective protector and antioxidant. The effects were closed to those of flavonoid silymarin-the classical hepatoprotector and antioxidant.
  7 2,301 79
Stability indicating studies on NMITLI 118RT+ (standardized extract of withania somnifera dunal)
Hafsa Ahmad, Kiran Khandelwal, Shakti Deep Pachauri, Rajender Singh Sanghwan, Anil Kumar Dwivedi
July-September 2014, 10(39):227-233
DOI:10.4103/0973-1296.137361  PMID:25210308
Background: Withania somnifera Dunal (Ashwagandha) is an Indian medicinal plant of great medicinal value; used in many clinically proven conditions. NMITLI-118RT+ is a candidate drug under a Council of Scientific and Industrial Research (CSIR) networking project. It is a chemotype of W. somnifera's root extract, which has been used for the present study. Objectives: The present investigation aims to develop and validate a simple isocratic reverse phase-high performance liquid chromatography (RP-HPLC) system for the detection and estimation of Withanolide A (marker compound) and its analytical application for stability indicating studies on NMITLI-118RT+. Material and Methods: A validated RP-HPLC method for Withanolide A was established on a Waters HPLC system and the same was used on NMITLI-118RT+ for quantification and fingerprinting purposes, and for establishing forced degradation, isothermal stress tests, and drug-excipient testing protocols as per International Conference on Harmonization (ICH) guidelines. Results: A validated method was established, which could detect the marker at a retention time of around 6.3 minutes, with a linearity range of 2-100 μg/mL, by varying the amounts of the said marker, which were estimated in four different batches of NMITLI-118RT+. Photostability as per ICH guidelines suggested a slight loss of the active constituent and maximum degradation was afforded with alkali followed by acid, and then peroxide, in the forced degradation studies. In the drug-excipient studies, the maximum amount of active constituent could be detected in the samples with ethyl cellulose and the least with hydroxy propyl cellulose. Conclusion: The method developed here was simple and rapid. The various stability indicating studies carried out in the present investigation would be useful for formulation development and were suggestive of deciding the recommended storage conditions for NMITLI-118RT+.
  6 3,780 109
Anticancer potential of Conium maculatum extract against cancer cells in vitro: Drug-DNA interaction and its ability to induce apoptosis through ROS generation
Jesmin Mondal, Ashis Kumar Panigrahi, Anisur Rahman Khuda-Bukhsh
August 2014, 10(39):524-533
DOI:10.4103/0973-1296.139792  PMID:25298670
Objective: Conium maculatum extract is used as a traditional medicine for cervix carcinoma including homeopathy. However, no systematic work has so far been carried out to test its anti-cancer potential against cervix cancer cells in vitro. Thus, in this study, we investigated whether ethanolic extract of conium is capable of inducing cytotoxicity in different normal and cancer cell lines including an elaborate study in HeLa cells. Materials and Methods: Conium's effects on cell cycle, reactive oxygen species (ROS) accumulation, mitochondrial membrane potential (MMP) and apoptosis, if any, were analyzed through flow cytometry. Whether Conium could damage DNA and induce morphological changes were also determined microscopically. Expression of different proteins related to cell death and survival was critically studied by western blotting and ELISA methods. If Conium could interact directly with DNA was also determined by circular dichroism (CD) spectroscopy. Results: Conium treatment reduced cell viability and colony formation at 48 h and inhibited cell proliferation, arresting cell cycle at sub-G stage. Conium treatment lead to increased generation of reactive oxygen species (ROS) at 24 h, increase in MMP depolarization, morphological changes and DNA damage in HeLa cells along with externalization of phosphatidyl serine at 48 hours. While cytochrome c release and caspase-3 activation led HeLa cells toward apoptosis, down-regulation of Akt and NFkB inhibited cellular proliferation, indicating the signaling pathway to be mediated via the mitochondria-mediated caspase-3-dependent pathway. CD-spectroscopy revealed that Conium interacted with DNA molecule. Conclusion: Overall results validate anti-cancer potential of Conium and provide support for its use in traditional systems of medicine.
  6 4,187 178
Hepatoprotective effects of Limonium tetragonum, edible medicinal halophyte growing near seashores
Min Hye Yang, Na-Hyun Kim, Jeong-Doo Heo, Sang Hyun Sung, Eun Ju Jeong
August 2014, 10(39):563-568
DOI:10.4103/0973-1296.139783  PMID:25298675
Background : During the process of hepatic fibrosis, the activation of hepatic stellate cells (HSCs) is responsible for the increased formation and reduced degradation of extracellular matrix in the liver. By employing the hepatic stellate cell line, HSC-T6, it was found that the methanol extract of Limonium tetragonum, a halophyte living in salt marsh near south and western seashores of Korea significantly inhibited the proliferation of HSC-T6 cells. Objective : In the present study, we attempted to investigate the antifibrotic effects of the mathanolic extract of L. tetragonum (MELT) in the activated HSC-T6 cells. Materials and Methods : The proliferation of HSC-T6 was stimulated by culturing environment or platelet-derived growth factor (PDGF-BB) insult, and then the inhibitory activities of MELT were measured. Results : It was found that MELT suppressed the proliferation of the activated HSC-T6 in concentration- and time-dependent manners. The increased collagen deposition in the activated HSC-T6 cells was also decreased by the treatment of MELT. The maximal dose of MELT, however, had little effect on primary cultured rat hepatocytes. Wlammatory cytokine, tumor necrosis factor alpha (TNF-α) produced by lipopolysaccharide-stimulated RAW264.7 macrophages was inhibited by MELT. Conclusion : Collectively, the above results demonstrated that MELT suppressed HSCs proliferation but not in hepatocytes, implying that L. tetragonum may be useful candidates for developing therapeutic agents for the prevention and treatment of hepatic fibrosis.
  6 2,535 98
Signal transducer and activators of transcription 3 regulates cryptotanshinone-induced apoptosis in human mucoepidermoid carcinoma cells
Hyun-Ju Yu, Chul Park, Sun-Ju Kim, Nam-Pyo Cho, Sung-Dae Cho
August 2014, 10(39):622-629
DOI:10.4103/0973-1296.139802  PMID:25298683
Background: Cryptotanshinone (CT) is a biologically active compound from the root of Salvia miltiorrhiza that has been reported to induce apoptosis in various cancer cell lines; but, it has not yet been fully explored in human mucoepidermoid carcinoma (MEC). Objective: Here, we demonstrated the apoptotic effects and its related mechanism in MC-3 and YD-15 human MEC cell lines. Materials and Methods: The effects of CT on apoptotic activity were evaluated by cell proliferation assay, Western blotting, 4'-6-diamidino-2-phenylindole staining, reverse transcription-polymerase chain reaction, and luciferase assay. Results: Our data show that CT treatment of MC-3 cells results in anti-proliferative and apoptotic activities in MC-3 and it is accompanied by a decrease in phosphorylation and dimerization of signal transducer and activators of transcription 3 (STAT3). CT decreased the expression levels of myeloid cell leukemia-1 (Mcl-1) and surviving, whereas Bcl-xL expression was not changed. CT clearly regulates survivin protein at a transcriptional level and alters Mcl-1 through proteasome-dependent protein degradation. In addition, CT-induced apoptotic cell death in YD-15, another human MEC cell line, was associated with the inhibition of STAT3 phosphorylation. Conclusion: These data suggest that CT could be a good apoptotic inducer through modification of STAT3 signaling in human MEC cell lines.
  5 2,306 78
New Sesquiterpene and Polymethoxy-Flavonoids from Artemisia annua L
Yang Chu, Hongbo Wang, Jing Chen, Yue Hou
July-September 2014, 10(39):213-216
DOI:10.4103/0973-1296.137359  PMID:25210306
Our previous study revealed that the polymethoxy-flavonoids, as main components of Artemisia annua, could improve the antimalarial activity of Artemisinin. Here, we described the isolation, elucidation, constituent analysis, flavonoids enrichment of the extracts of A. annua. A total of 20 compounds were isolated including a new sesquiterpene (compound 12) and five (1, 5, 6, 7, 15) afforded for the first time from A. annua. The elucidation of eight flavonoids may be a useful phytochemical data and chemical foundation for further mechanism studies on improving the anti-malarial action of artemisinin. Furthermore, the antitumor activities of the compounds were assayed using four different kinds of human cancer cell lines.
  5 3,301 116
Fast analysis of principal volatile compounds in crude and processed Atractylodes macrocephala by an automated static headspace gas chromatography-mass spectrometry
Jida Zhang, Gang Cao, Yunhua Xia, Chengping Wen, Yongsheng Fan
July-September 2014, 10(39):249-253
DOI:10.4103/0973-1296.137364  PMID:25210311
Objective: Atractylodes macrocephala, a famous herbal medicine, is used extensively in the practice of Traditional Chinese Medicine (TCM). Processing procedure is a common approach that usually occurs before A. macrocephala is prescribed. This paper describes a sensitive and specific assay for the determination of principal volatile compounds in crude and processed A. macrocephala. Materials and Methods: The present study concentrated on the development of a static headspace gas chromatography-mass spectrometry (SHS-GC/MS) for separating and identifying of volatile compounds from crude and processed A. macrocephala samples. Results: The results showed that the volatile oil in crude and processed A. macrocephala was markedly quantitatively and qualitatively different. Processing resulted in the reduction of volatile oil contents and variation of chemical compositions in A. macrocephala. Conclusion: The proposed method proved that SHS-GC/MS is rapid and specific, and should also be useful for evaluating the quality of crude and processed medicinal herbs.
  5 3,196 134
Methyl protodioscin induces G2/M cell cycle arrest and apoptosis in A549 human lung cancer cells
Yang Bai, Xiao-Yuan Qu, Jun-Qiang Yin, Liangcai Wu, Hong Jiang, Han-Wu Long, Qiang Jia
July-September 2014, 10(39):318-324
DOI:10.4103/0973-1296.137373  PMID:25210320
Background: Methyl protodioscin (MPD) is a furostanol bisglycoside with antitumor properties. It has been shown to reduce proliferation, cause cell cycle arrest. Objective: The present study elucidates the mechanism underlying MPD's apoptotic effects, using the A549 human lung cancer cell line. Materials and Methods: The human pulmonary adenocarcinoma cell line A549 was obtained from the Cell Bank of the Animal Experiment Center, North School Region, Sun Yat-Sen University. All of the cells were grown in RPMI 1640 supplemented with 10% fetal calf serum (Hyclone, Logan, UT, USA), penicillin (10,000 U/l), and streptomycin (100 mg/l) at 37°C in a 5% CO 2 humidified atmosphere. The induction of apoptosis was observed in flow cytometry and fluorescent staining experiments. Results: MPD showed growth inhibitory effects in A549 cells in a dose- and time-dependent manner. The significant G2/M cell cycle arrest and apoptotic effect were also seen in A549 cells treated with MPD. MPD-induced apoptosis was accompanied by a significant reduction of mitochondrial membrane potential, release of mitochondrial cytochrome c to cytosol, activation of caspase-3, downregulation of Bcl-2, p-Bad, and upregulation of Bax. Conclusion: Our results show that the induction of apoptosis by MPD involves multiple molecular pathways and strongly suggest that Bcl-2 family proteins signaling pathways. In addition, mitochondrial membrane potential, mitochondrial cytochrome c and caspase-3 were also closely associated with MPD-induced apoptotic process in human A549 cells.
  5 3,409 95
Phytochemical compositions of volatile oil from Blumea balsamifera and their biological activities
Zhi-long Jiang, Yan Zhou, Wei-chen Ge, Ke Yuan
July-September 2014, 10(39):346-352
DOI:10.4103/0973-1296.137377  PMID:25210324
Background: This research, extract compositions of volatile oil in Blumea balsamifera (Linn.) DC by steam distillation method. The gas chromatograph-mass spectrometer technology (GC-MS) was used to separate and determine chemical components of volatile oil. Meanwhile, the percentage of the chemical components was determined in the volatile oil by peak area normalization method. At the same time, we determine and evaluate antitumor, antibacterial, and antioxidation activities of chemical components of volatile oil in Blumea balsamifera of the aromatic plant. Materials and Methods: We screened the cytotoxicity of volatile oil in Blumea balsamifera by using prawn larva and by prawn-lethal bioactivity experiment. Use the slanting test tube method to evaluate the antibacterial activity of volatile oil in Blumea balsamifera for eight kinds of plant pathogenic fungi. Taking Trolox as the contrast, the research uses DPPH method to study the radical-scavenging function (IC 50 ) of the volatile oil in Blumea balsamifera. Results: The results show that 42 kinds of compounds are separated from volatile oil of Blumea balsamifera. The appraised components take up 97.65% of total peak area. The volatile oil in Blumea balsamifera mainly contains sesquiterpenoids. The results also show that it has relatively strong activity of antitumor and anti-plant pathogenic fungi and some antioxidation activity. Conclusion: This research provided the reference data for further development of this natural resource, and at the same time, we understood more of the chemical components of volatile oil and bioactivity of this aromatic plant.
  5 5,424 253
Simultaneous determination of esculetin, quercetin-3-O-β-D-glucuronide, quercetin-3-O-β -D-glucuronopyranside methyl ester and quercetin in effective part of Polygonum Perfoliatum L. using high performace liquid chromatography
Dongsheng Fan, Yang Zhao, Xin Zhou, Xiaojian Gong, Chao Zhao
July-September 2014, 10(39):359-366
DOI:10.4103/0973-1296.137379  PMID:25210326
Objective: In the present study, a high performance liquid chromatography (HPLC) coupled with photodiode array detection was developed for simultaneous quantitation of esculetin, quercetin-3-O-β-D-glucuronide, quercetin-3-O-β-D- glucuronopyranoside methyl ester and quercetin in Polygonum perfoliatum L. Materials and Methods: The chromatographic separations were performed on a reversed-phase C 18 column using a mobile phase composed of acetonitrile -0.5% aqueous acetic acid with gradient elution. The calibration curves for the analytes demonstrated good linearities within the investigated ranges. The satisfactory intra- and inter-day precision, repeatability and stability of the developed analytical method were shown in the method validation procedure. The recoveries of the established method ranged from 95.76 to 102.10% for all the analytes. Results: This proposed method was successfully applied for simultaneous quantification of the four compounds in effective part of Polygonum perfoliatum L. from different regions. Hierarchical clustering analysis (HCA) and principal components analysis (PCA) were performed to characterize and classify the samples based on the contents of the four compounds in Polygonum perfoliatum L. Conclusion: The established HPLC method combined with chemometric approaches was proven to be useful and efficient for quality control of Polygonum perfoliatum L.
  5 3,221 88
Multiple shoot regeneration and effect of sugars on growth and nitidine accumulation in shoot cultures of Toddalia asiatica
Chinthala Praveena, Ciddi Veeresham
August 2014, 10(39):480-486
DOI:10.4103/0973-1296.139777  PMID:25298663
Background: Toddalia asiatica (Rutaceae) is an important medicinal plant in traditional medicinal system of India and China. Nitidine production from callus cultures of the plant had been investigated, but in vitro multiplication and secondary metabolite production from shoot cultures is not reported. Objective: The aim of the present work is to establish protocol for in vitro multiple shoot regeneration of T. asiatica and to investigate the secondary metabolite, nitidine production from the shoot cultures. Materials and Methods: Different explants were used for shoot regeneration on MS supplemented with benzyl adenine (BA) either alone or in combination with naphthalene acetic acid (NAA) in different combinations. Effect of different sugars and different concentrations of sucrose on biomass accumulation in shoot cultures in liquid medium was investigated. For in vitro rooting, shoots culture were inoculated to half strength MS medium supplemented with different concentrations of indole butyric acid. Quantitative analysis of shoot culture extracts was done for estimation of nitidine by HPTLC. Results: Shoot cultures were successfully initiated and established from nodal and shoot tip explants on MS medium supplemented with benzyl adenine and sucrose (3% w/v). Sucrose at a concentration of 3 % w/v was found to be optimum for growth and biomass accumulation. In vitro rooting of shoots was achieved on half strength MS medium supplemented with indole butyric acid 3 mg/l. Investigation of secondary metabolite production ability of the in vitro regenerated shoot cultures revealed their ability to biosynthesize nitidine. Conclusion: Shoot cultures were established and nitidine production has been observed.
  4 3,310 134
Development and validation of an UPLC-ESI-MS/MS method for determination of dehydroevodiamine, limonin, evodiamine, and rutaecarpine in Evodiae Fructus
Yang Zhao, Yunling Zhao, Xin Zhou, Xiaojian Gong
July-September 2014, 10(39):374-383
DOI:10.4103/0973-1296.137381  PMID:25210328
Objective: Evodiae Fructus (EF), one of the most widely used traditional Chinese medicines, mainly consists of alkaloids, is widely used for the treatments of headache and gastrointestinal disorders. In this study, a sensitive and reliable UPLC-ESI-MS/MS method was developed for qualitative determination of dehydroevodiamine, limonin, evodiamine, and rutaecarpine. Materials and Methods: Chromatographic separations were accomplished on a Phenomenex Kinetex XB-C18 column (2.1 × 150 mm, 1.7 μm) by using a gradient elution profile with a mobile phase consisting of 0.5% formic acid in water (A) and acetonitrile (B). Detection was performed using multiple reactions monitoring mode under ESI in the positive ion mode. Results: The results showed good linearity over the investigated concentration ranges (R2 >0.9900) for the analytes. The limit of quantitations (LOQs) were 6.88 ng/mL for dehydroevodiamine, 18.6 ng/mL for limonin, 6.24 ng/mL for evodiamine, and 2.56 ng/mL for rutaecarpine, respectively. Intraday and interday precisions (relative standard deviations, %) were <5% and accuracies ranged from 92% to 106%. Conclusion: The validated method was successfully applied to assay the contents of the four compounds in EF samples from different regions, with which just 10 min was needed to analyze each sample.
  4 3,074 112
Optimization of simultaneous ultrasonic-assisted extraction of water-soluble and fat-soluble characteristic constituents from Forsythiae Fructus Using response surface methodology and high-performance liquid chromatography
Yong-Gang Xia, Bing-You Yang, Jun Liang, Di Wang, Qi Yang, Hai-Xue Kuang
July-September 2014, 10(39):292-303
DOI:10.4103/0973-1296.137370  PMID:25210317
Background: The compounds (+)-pinoresinol-β-glucoside (1) forsythiaside, (2) phillyrin (3) and phillygenin (4) were elucidated to be the characteristic constituents for quality control of Forsythiae Fructus extract by chromatographic fingerprint in 2010 edition of Chinese Pharmacopoeia due to their numerous important pharmacological actions. It is of great interest to extract these medicinally active constituents from Forsythiae Fructus simultaneously. Materials and Methods: In this study, a new ultrasound-assisted extraction (UAE) method was developed for the simultaneous extraction of biological components 1-4 in Forsythiae Fructus. The quantitative effects of extraction time, ratio of liquid to solid, extraction temperature, and methanol concentration on yield of these four important biological constituents from Forsythiae Fructus were investigated using response surface methodology with Box-Behnken design. The compounds 1-4 extracted by UAE were quantitative analysis by high-performance liquid chromatography-photodiode array detect (HPLC-PAD), and overall desirability (OD), the geometric mean of the contents of four major biological components, was used as a marker to evaluate the extraction efficiency. Results: By solving the regression equation and analyzing 3-D plots, the optimum condition was at extraction temperature 70°C, time 60 min, ratio of liquid to solid 20, and methanol concentration 76.6%. Under these conditions, extraction yields of compounds 1-4 were 2.92 mg/g, 52.10 mg/g, 0.90 mg/g and 0.57 mg/g, respectively, which were in good agreement with the predicted OD values. In order to achieve a similar yield as UAE, soxhlet extraction required at least 6 h and maceration extraction required much longer time of 24 h. Established UAE method has been successfully applied to sample preparation for the quality control of Forsythiae Fructus. Additionally, a quadrupole time-of-flight mass spectrometry was applied to the structural confirmation of analytes from the complex matrices acquired by UAE. Conclusion: The results indicated that UAE is an effective alternative method for extracting bioactive constituents, which may facilitate a deeper understanding of the extract of active constituents in Forsythiae Fructus from the raw material to its extract for providing the theoretical references.
  4 3,772 145
Comparison of toad skins Bufo bufo gargarizans Cantor from different regions for their active constituents content and cytotoxic activity on lung carcinoma cell lines
Congyan Liu, Wei Cao, Yan Chen, Ding Qu, Jing Zhou
July-September 2014, 10(39):207-212
DOI:10.4103/0973-1296.137358  PMID:25210305
Background: The skin of Bufo bufo gargarizans Cantor, rich in bufadienolides, peptides, and alkaloids, has approved pharmacological activity for preliminary anti-liver and lung tumor treatment. However, few studies have systematically focused on the influence of the producing regions on the content and antitumor activity of the active constituents in toad skins. Objective: This study aims to compare toad skins obtained from six different regions in China (Jiangsu, Anhui, Henan, Hebei, Jiangxi, and Shandong province) for their bufadienolide and alkaloid content, and their cytotoxic activity on two lung carcinoma cell lines (SPC-A-1 cells and A549 cells). Materials and Methods: High performance liquid chromatography (HPLC) was used to quantificationally determine four bufadienolides, which included bufotalin, bufalin, cinobufagin, and resibufogenin in toad skins, from six different regions, respectively. In addition, an ultraviolet (UV) spectrophotometer was also employed to identify the content of the total alkaloids using 5-hydroxytryptamine (5-HT) as the reference substance. An MTT assay was performed to compare the antiproliferative effects of the toad skins' ethanolic extracts from the different regions against SPC-A-1 and A549 cells. Results: In this study, the toad skins from Jiangsu province had the highest amount of bufadienolides (472.6 μg/g crude drug) and alkaloids (1.51 mg/g crude drug). Meanwhile, according to the extract, it exhibited the strongest cytotoxic effect against the lung carcinoma cell line (SPC-A-1 cells and A549 cells) with IC 50 values of 24.82 ± 0.76 and 23.77 ± 0.63 μg crude drug/mL, respectively. Conclusion: The toad skins that originated from the Jiangsu province, have comparatively greater advantages over samples from other regions as far as active constituent content and potential anti-lung cancer activity is concerned, suggesting that it can be a promising chemotherapeutic agent in lung cancer therapy, in further studies.
  4 3,377 119
Lactobacilli-fermented Hwangryunhaedoktang has enhanced anti-inflammatory effects mediated by the suppression of MAPK signaling pathway in LPS-stimulated RAW 264.7 cells
You-Chang Oh, Yun Hee Jeong, Won-Kyung Cho, Kwang Jin Lee, Taesoo Kim, Jin Yeul Ma
August 2014, 10(39):645-654
DOI:10.4103/0973-1296.139815  PMID:25298686
Background: Hwangryunhaedoktang (HR) has been traditionally used in oriental medicine as a drug for the treatment of melena, hemoptysis, and apoplexy. Objective: We investigated whether HR and lactobacilli-fermented HRs have an inhibitory effect on the production of proinflammatory mediators in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. Materials and Methods: The investigation was focused on whether HR and fermented HRs could inhibit the production of prostaglandin (PG)E 2, nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin (IL)-6, cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS) and mitogen-activated protein kinases (MAPKs) in LPS-stimulated RAW 264.7 cells. Results: We found that HR weakly inhibited various inflammatory mediators induced by LPS. However, fermentation with lactobacilli significantly increased the inhibitory effect of HR on most of the inflammatory mediator expression. Furthermore, fermented HRs exerted a stronger inhibitory effect on MAPKs phosphorylation than that by non-fermented HR. Conclusions: These results suggest that lactobacilli-fermented HRs contains elevated potent anti-inflammatory activity that is mediated by inhibiting MAPKs pathway in macrophages.
  4 2,385 81
Anti-inflammatory effect of Artemisiae annuae herba in lipopolysaccharide-stimulated RAW 264.7 Cells
You-Chang Oh, Yun Hee Jeong, Taesoo Kim, Won-Kyung Cho, Jin Yeul Ma
August 2014, 10(39):588-595
DOI:10.4103/0973-1296.139793  PMID:25298679
Background: Artemisiae annuae herba (AAH) has been traditionally used as a drug for the treatment of malaria, heat stroke, bacterial infection, and fever in East-Asia. Although AAH has been used for the treatment of inflammation-related symptoms, the underlying mechanism of antiinflammatory activity of AAH is still unknown. Objective: We investigated whether AAH have an inhibitory effect on the production of pro-inflammatory mediators in lipopolysaccharide-stimulated RAW 264.7 macrophage cells. Materials and Methods: The investigation was forced on the inhibitory effect of AAH on the production of tumor necrosis factor (TNF)-α, interleukin (IL)-6, nitric oxide (NO), and inducible NO synthase (iNOS) in macrophages. Furthermore, we examined the effect of AAH on the activation of nuclear factor kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) pathways. Results: We found that AAH suppresses NO production and TNF-α, IL-6, and iNOS gene expression. Moreover, AAH inhibited the nuclear translocation of p65 and IκBα degradation in NF-κB pathway and decreased the extracellular signal-regulated kinase, p38, c-Jun NH 2 -terminal kinase phosphorylation in MAPK signaling pathway. Conclusions: Consequently, these results indicate that AAH contains antiinflammatory activity and this effect is derived from the repression on the activation of NF-κB and MAPKs pathways. We first demonstrated that antiinflammatory effect of AAH and its underlying mechanism in macrophage cells.
  3 2,686 103
Optimization of the extraction process for the seven bioactive compounds in Yukmijihwang-tang, an herbal formula, using response surface methodology
Jung-Hoon Kim, Hyeun-Kyoo Shin, Chang-Seob Seo
August 2014, 10(39):606-613
DOI:10.4103/0973-1296.139798  PMID:25298681
Background: Yukmijihwang-tang (YJT) contains multiple bioactive compounds. Heat-reflux extraction was employed and optimized for the extraction of the bioactive compounds in YJT. Objective: The determination of optimal conditions with maximum yields of bioactive compounds, gallic acid, 5-hydroxymethylfurfural, morroniside, loganin, paeoniflorin, benzoic acid and paeonol, in YJT. Materials and Methods: The extraction ratio (ratio of water to herbal formula), extraction time and extraction number were set as individual values and the yields of the seven compounds were the response values that were optimized with a Box-Behnken design. Results: The optimal conditions obtained from response surface methodology (RSM) were 1:11.99 for the extraction ratio, 94.53 min for the extraction time and 2.21 for the extraction number. Under the optimal conditions, the response value of the experiment closely agreed with the predicted response value. Conclusions: The result suggests that RSM is successfully applied for optimizing the extraction of the marker compounds in YJT.
  3 2,891 155
A new mulinane diterpenoid from the cushion shrub Azorella compacta growing in Perú
Francisco Salgado, Carlos Areche, Beatriz Sepúlveda, Mario J Simirgiotis, Fátima Cáceres, Cristina Quispe, Lina Quispe, Teresa Cano
August 2014, 10(39):543-548
DOI:10.4103/0973-1296.139807  PMID:25298672
Background: Azorella compacta is a rare yellow-green compact resinous cushion shrub growing from the high Andes of southern Perϊ to northwestern Argentina, and which is a producer of biologically active and unique diterpenoids. Objective: This study investigated the secondary metabolites present in a Peruvian sample of Azorella compacta and the evaluation of gastroprotective activity of the isolated compounds in a gastric- induced ulcer model in mice. Material and Methods: Six secondary metabolites (diterpenoids 1-6) present in the dichloromethane (DCM) extract of A. compacta growing in Perϊ were isolated by a combination of Sephadex LH-20 permeation and silica gel chromatography and their chemical structures were elucidated by spectroscopic methods (NMR) and molecular modeling. The gastroprotective activity of the new compound 1 was evaluated on the HCl/EtOH-induced gastric lesion model in mice and compared to the activity showed by the known compounds. Results: A new mulinane diterpene along with five known diterpenoids have been isolated from a Peruvian  sample of A. compacta and the gastroprotective results show that compound 1 is less active than the other known mulinane diterpenoids isolated. Conclusions: A. compacta growing in Perϊ showed the presence of the new mulinane 1, which was poorly active in the HCl/EtOH-induced gastric lesion model in mice. Indeed, the activity was lower than other diterpenoids (2-6) showing an oxygenated function at C-16 or/and C-20, which confirm the role of an oxygenated group (OH or carboxylic acid) for the gastroprotective activity of mulinane compounds.
  3 2,617 92
Effect of three fatty acids from the leaf extract of Tiliacora triandra on P-glycoprotein function in multidrug-resistant A549RT-eto cell line
Chutima Kaewpiboon, Pakorn Winayanuwattikun, Tikamporn Yongvanich, Preecha Phuwapraisirisan, Wanchai Assavalapsakul
August 2014, 10(39):549-556
DOI:10.4103/0973-1296.139779  PMID:25298673
Background: Cancer cells have the ability to develop resistance to chemotherapy drugs, which then leads to a reduced effectiveness and success of the treatment. Multidrug resistance (MDR) involves the resistance in the same cell/tissue to a diverse range of drugs of different structures. One of the characteristics of MDR is an overexpression of P-glycoprotein (P-gp), which causes the efflux of the accumulated drug out of the cell. The MDR human non-small cell lung carcinoma cell line with a high P-gp expression level (A549RT-eto) was used to investigate the bioactive compounds capable of reversing the etoposide resistance in this cell line. Materials and Methods: The leaves of Tiliacora triandra were sequentially extracted with hexane, dichloromethane, methanol and water. Only the hexane extract reduced the etoposide resistance of the A549RT-eto cell line, and was further fractionated by column chromatography using the TLC-pattern and the restoration of etoposide sensitivity as the selection criteria. Results: The obtained active fraction (F22) was found by nuclear magnetic resonance and gas chromatography-mass spectroscopy analyses to be comprised of a 49.5:19.6:30.9 (w/w/w) mixture of hexadecanoic: octadecanoic acid: (Z)-6-octadecenoic acids. This stoichiometric mixture was recreated using pure fatty acids (MSFA) and gave a similar sensitization to etoposide and enhanced the relative rate of rhodamine-123 accumulation to a similar extent as F22, supporting the action via reducing P-gp activity. In contrast, the fatty acids alone did not show this effect. Conclusion: This is the first report of the biological activity from the leaves of T. triandra as a potential source of a novel chemosensitizer.
  3 2,443 86
Rapid and undamaged analysis of crude and processed Radix Scrophulariae by Fourier transform infrared spectroscopy coupled with soft independent modeling of class analogy
Huiping Zhu, Gang Cao, Hao Cai, Baochang Cai, Jue Hu
July-September 2014, 10(39):265-270
DOI:10.4103/0973-1296.137366  PMID:25210313
Objective: The main objective of this work is to determine the feasibility of identification of crude and processed Radix Scrophulariae using the Fourier transform infrared spectroscopy couple with soft independent modeling of class analogy (FT-IR-SIMCA). Materials and Methods: A total of 50 different crude Radix Scrophulariae was used to product processed ones. The spectra were acquired by FT-IR spectroscopy using a diffuse reflectance fiber optic probe. For the multivariate analysis, SIMCA was used. Results showed that FT-IR-SIMCA was useful to discriminate the processed Radix Scrophulariae samples from crude samples. These samples could be successfully classified by SIMCA. Results: In all cases, the recognition and rejection rates were 97.8% and 100%, respectively. When testing with the blind sample that was picked out from the chosen samples, the accuracy was up to 90%. Conclusion: It means that the methodology is capable of accurately separating processed Radix Scrophulariae from crude samples.
  3 2,542 57
High performance liquid chromatography profiling of health-promoting phytochemicals and evaluation of antioxidant, anti-lipoxygenase, iron chelating and anti-glucosidase activities of wetland macrophytes
Keng-Fei Ooh, Hean-Chooi Ong, Fai-Chu Wong, Nam-Weng Sit, Tsun-Thai Chai
August 2014, 10(39):443-455
DOI:10.4103/0973-1296.139767  PMID:25298659
Background: The phytochemistry and bioactivity of wetland macrophytes are underexplored. Plants are known as the natural sources of phytochemical beneficial to health. Objective: The objective of this study is to analyze the phytochemical profiles and bioactivities of 10 extracts prepared from different plant parts of wetland macrophytes Hanguana malayana, Ludwigia adscendens and Monochoria hastata. Materials and Methods: High performance liquid chromatography (HPLC) was used to analyze the phytochemical profile of the extracts. Antioxidant assay such as 2,2-diphenyl-1-picrylhydrazyl, nitric oxide (NO) radical scavenging activity and ferric reducing antioxidant power were performed. Bioactivity assays carried out were anti-lipoxygenase, anti-glucosidase, and iron chelating. Results: Leaf extract of L. adscendens had the highest 2,2-diphenyl-1-picrylhydrazyl (half of maximal effective concentration [EC 50 ] =0.97 mg/mL) and NO (EC 50 = 0.31 mg/mL) scavenging activities. The extract also exhibited the highest iron chelating (EC 50 = 3.24 mg/mL) and anti-glucosidase (EC 50 = 27.5 μg/mL) activities. The anti-glucosidase activity of L. adscendens leaf extract was comparable or superior to those of acarbose, myricetin and quercetin. Correlation between iron chelating and radical scavenging activities among the extracts implies the presence of dual-function phytoconstituents with concurrent iron chelating and radical scavenging activities. HPLC analysis revealed the presence of p-coumaric acid (p-CA), gallic acid (GA) and myricetin in all or most extracts. M. hastata fruit and leaf extracts had the highest p-hydroxybenzoic acid content. Antioxidant and anti-glucosidase activities of the extracts were correlated with p-CA, GA, and myricetin contents. Conclusion: Our study demonstrated that wetland macrophytes H. malayana, L. adscendens and M. hastata are potential sources of health-promoting phytochemicals with potent therapeutically-relevant bioactivities.
  3 3,176 220
Inhibition of inducible nitric oxide synthase and osteoclastic differentiation by Atractylodis Rhizoma Alba extract
Sung-Ho Choi, Sung-Jin Kim
August 2014, 10(39):494-500
DOI:10.4103/0973-1296.139780  PMID:25298665
Background: Atractylodis Rhizoma Alba (ARA) has been used in Korean folk medicine for constipation, dizziness, and anticancer agent. In the present study, we performed to test whether the methanolic extract of ARA has antioxidant and antiosteoclastogenesis activity in RAW 264.7 macrophage cells. Materials and Methods: Antioxidant capacities were tested by measuring free radical scavenging activity, nitric oxide (NO) levels, reducing power, and inducible nitric oxide synthase (iNOS) expression in response to lipopolysaccharides (LPS). Antiosteoclastogenesis activity was evaluated by performing tartrate-resistant acid phosphatase assay in RAW 264.7 macrophage cells. Results: The extract exerted significant 1,1-diphenyl-2-picrylhydrazyl and NO radical scavenging activity, and it exerted dramatic reducing power. Induction of iNOS and NO by LPS in RAW 264.7 cells was significantly inhibited by the extract, suggesting that the ARA extract inhibits NO production by suppressing iNOS expression. Strikingly, the ARA extracts substantially inhibited the receptor activator of NF-κB ligand-induced osteclastic differentiation of LPS-activated RAW 264.7 cells. The ARA extract contains a significant amount of antioxidant components, including phenolics, flavonoids and anthocyanins. Conclusion: These results suggest that the methanolic extract of ARA exerts significant antioxidant activities potentially via inhibiting free radicals and iNOS induction, thereby leading to the inhibition of osteoclastogenesis.
  3 3,055 204
Novel characterization of Radix Angelicae Dahuricae before and after the sulfur-fumigation process by combining high performance liquid chromatographic fingerprint and multi-ingredients determination
Xiao Liu, Jingjing Liu, Hao Cai, Songlin Li, Xiaoqing Ma, Yajing Lou, Kunming Qin, Hongyue Guan, Baochang Cai
July-September 2014, 10(39):338-345
DOI:10.4103/0973-1296.137376  PMID:25210323
Background: Harmful sulfur-fumigation processing method is abused during Radix Angelicae Dahuricae preparation. However, the analytical technique characterizing Radix Angelicae Dahuricae before and after the sulfur-fumigation process is absent. Materials and Methods: The high performance liquid chromatography (HPLC) technique was adopted to develop methods combining finger-print analysis and multi-ingredients simultaneous determination for quality evaluation of Radix Angelicae Dahuricae before and after the sulfur-fumigation process. The chromatographic fingerprint method was established for qualitative analysis coupled with statistical cluster analysis basing on Euclidean distance. Additionally, a determination method was developed for quantitative analysis, which was able to assay the concentrations of the major coumarins including imperatorin, isoimperatorin, xanthotoxin, xanthotoxol, isoimpinellin, oxypeucedanin, and bergapten in Radix Angelicae Dahuricae simultaneously. The separations of the two methods were both achieved on a Hypersil octadecylsilyl C 18 column (250 mm × 4.6 mm, 5 μm) at 35°C under different strategic gradient elution programs. The detection wavelength was set at 254 nm all the time. Method validation data indicated that the methods were both reliable and applicable. They were then used to assay different Radix Angelicae Dahuricae samples collected from good agricultural practice (GAP) bases and local herbal markets. Results: The successful application demonstrated that the combination of HPLC fingerprint and simultaneous quantification of multi-ingredients offers an efficient approach for quality evaluation of Radix Angelicae Dahuricae before and after the sulfur-fumigation process. Conclusion: In order to discriminate Radix Angelicae Dahuricae before and after the sulfur-fumigation process, oxypeucedanin, and xanthotoxol were the most sensitive biomarkers and should be determined.
  3 2,821 83
A rapid identification of four medicinal chrysanthemum varieties with near infrared spectroscopy
Bangxing Han, Hui Yan, Cunwu Chen, Houjun Yao, Jun Dai, Naifu Chen
July-September 2014, 10(39):353-358
DOI:10.4103/0973-1296.137378  PMID:25210325
Background: For genuine medicinal material in Chinese herbs; the efficient, rapid, and precise identification is the focus and difficulty in the filed studying Chinese herbal medicines. Chrysanthemum morifolium as herbs has a long planting history in China, culturing high quality ones and different varieties. Different chrysanthemum varieties differ in quality, chemical composition, functions, and application. Therefore, chrysanthemum varieties in the market demands precise identification to provide reference for reasonable and correct application as genuine medicinal material. Materials and Methods: A total of 244 batches of chrysanthemum samples were randomly divided into calibration set (160 batches) and prediction set (84 batches). The near infrared diffuses reflectance spectra of chrysanthemum varieties were preprocessed by first order derivative (D1) and autoscaling and was built model with partial least squares (PLS). Results: In this study of four chrysanthemum varieties identification, the accuracy rates in calibration sets of Boju, Chuju, Hangju, and Gongju are respectively 100, 100, 98.65, and 96.67%; while the accuracy rates in prediction sets are 100% except for 99.1% of Hangju. Conclusion: The research results demonstrate that the qualitative analysis can be conducted by machine learning combined with near infrared spectroscopy (NIR), which provides a new method for rapid and noninvasive identification of chrysanthemum varieties.
  3 2,777 105
Antitumor effects of dammarane-type saponins from steamed Notoginseng
Fan He, Yan Ding, Chun Liang, Seok Bean Song, De-Qiang Dou, Gyu Yong Song, Young Ho Kim
July-September 2014, 10(39):314-317
DOI:10.4103/0973-1296.137372  PMID:25210319
Six dammarane-type saponins were extracted from steamed Panax notoginseng. Their chemical structures were identified spectroscopically as ginsenosides Rh 1 (1), Rg 1 (2), 20 (S)-Rg 3 (3), 20 (R)-Rg 3 (4), Rb 3 (5), and Rb 1 (6). Compounds (0.1-10 μM) were tested for inhibition of tumor necrosis factor-α (TNF)-induced nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) luciferase reporter activity using a human kidney 293T cell-based assay. Ginsenoside Rb 3 (5) showed the most significant activity with an IC 50 of 8.2 μM. This compound also inhibited the induction of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) messenger Ribonucleic acid (mRNA) in a dose-dependent manner after HepG2 cells had been treated with TNF-α (10 ng/mL).
  3 5,179 86
Induction of apoptosis by Cordyceps militaris fraction in human chronic myeloid leukemia K562 cells involved with mitochondrial dysfunction
Tian Tian, Liyan Song, Qin Zheng, Xianjing Hu, Rongmin Yu
July-September 2014, 10(39):325-331
DOI:10.4103/0973-1296.137374  PMID:25210321
Background: Cordyceps militaris is widely used for various ethno medical conditions including cancer and inflammation complications in traditional Chinese medicine. Objective: To investigate the in vitro antitumor activity of Cordyceps militaris fraction (CMF) and the molecular mechanism underlying the apoptosis it induces in human chronic myeloid leukemia K562 cells. Materials and Methods: CMF was prepared according to our previous report. Cell viability was assessed by MTT assay. The rate of apoptosis, distribution of cell cycle and loss of mitochondrial membrane potential were measured by flow cytometry. Caspase activities were analyzed by Western blot and oxygen consumption rate was recorded using the Oxytherm system. Results: The results demonstrated that CMF triggered growth inhibition in K562 cells with only minor toxicity on a normal human cell line and inhibited the proliferation of K562 cells in a dose- and time-dependent manner with IC 50 value of 34.1 ± 2.0 μg/ml after 48 h incubation. This most likely resulted from cell cycle arrest at the S phase and the induction of apoptosis. In addition, CMF induced activation of caspase-3 and subsequent cleavage of poly ADP-ribose polymerase (PARP). The caspase signals may originate from mitochondrial dysfunction, which was supported by the finding of decreased mitochondria transmembrance potential and the lower oxygen consumption rate. Conclusion: CMF possessed the in vitro antitumor effect on K562 cells and CMF-induced apoptosis might be involved by the mitochondrial dysfunction and valuable to research and develop as a potential antitumor agency.
  2 3,663 71
Enhanced dissolution and stability of Tanshinone IIA base by solid dispersion system with nano-hydroxyapatite
Yan-rong Jiang, Zhen-hai Zhang, Sai-yan Huang, Yan Lu, Tian-tian Ma, Xiao-bin Jia
July-September 2014, 10(39):332-337
DOI:10.4103/0973-1296.137375  PMID:25210322
Background: Tanshinone IIA (TSIIA) exhibits a variety of cardiovascular effects; however, it has low solubility in water. The preparation of poorly soluble drugs for oral delivery is one of the greatest challenges in the field of formulation research. Among the approaches available, solid dispersion (SD) technique has proven to be one of the most commonly used these methods for improving dissolution and bioavailability of drugs, because of its relative simplicity and economy in terms of both preparation and evaluation. Objective: This study was aimed at investigating the dissolution behavior and physical stability of SDs of TSIIA by employing nano-hydroxyapatite (n-HAp). Materials and Methods: The TSIIA SDs was prepared to use a spray-drying method. First, an in vitro dissolution test was performed to assess dissolution characteristics. Next, a set of complementary techniques (differential scanning calorimetry, scanning electron microscopy, X-ray powder diffraction, and Fourier transform infrared spectroscopy) was used to monitor the physicochemical properties of the SDs. The SDs was stored at 40°C/75% relative humidity for 6 months, after which their stability was assessed. Results: TSIIA dissolution remarkably improved because of the formulation of the SDs with n-HAp particles. Comparisons with the corresponding physical mixtures revealed changes in the SDs and explained the formation of the amorphous phase. In the stability test, virtually no time-dependent decrease was observed in either in vitro drug dissolution or drug content. Conclusion: SD formulation with n-HAp may be a promising approach for enhancing the dissolution and stability of TSIIA.
  2 2,849 67
Analysis of the influence of sulfur-fumigation on the volatile components of Angelicae sinensis Radix by comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry
Gang Cao, Hao Cai, Yajing Lou, Sicong Tu, Xiao Liu, Kunming Qin, Baochang Cai
July-September 2014, 10(39):304-313
DOI:10.4103/0973-1296.137371  PMID:25210318
Background: Sulfur-fumigation of Angelicae sinensis Radix causes changes in the structure and composition of volatile components. These changes alter the curative effect and the quality of A. sinensis Radix. Materials and Methods: In this study, comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC × GC-TOFMS) was employed to investigate the influence of sulfur-fumigation on the volatile components, and to characterize and quantify the chemical composition of the volatile oil of A. sinensis Radix. Results: The present study has shown that sulfur-fumigated A. sinensis Radix samples had significant loss of the main active compounds and a more destructive fingerprint profile compared to non-fumigated samples. Conclusion: From this study, it can be concluded that the combination of GC × GC and TOFMS has potential as a quality monitoring tool in herbal medicine and food processing industries.
  2 2,770 80
Pharmacokinetics and tissue distribution study of tanshinone IIA after oral administration of Bushen Huoxue Qubi granules to rats with blood stasis syndrome
Yuanqing Wang, Jianye Yan, Shunxiang Li, Xiong Cai, Wei Wang, Kun Luo, Dan Huang, Jiesheng Gao
July-September 2014, 10(39):285-291
DOI:10.4103/0973-1296.137369  PMID:25210316
Background: Bushen Huoxue Qubi (BHQ) granules, a traditional Chinese medicine preparation, has been clinically used for the treatment of the blood stasis syndrome. Objective: The main objective is to investigate whether the diseased condition would alter the pharmacokinetics and tissue distribution of tanshinone IIA in BHQ, which was given orally to the acute blood stasis rats. Materials and Methods: The main bioactive constituent in BHQ, tanshinone IIA, was measured in the plasma and tissues of animals by the high performance liquid chromatography with ultraviolet detection. The analysis was successfully performed on an Agilent TC-C 18 column (250 × 4.6 mm I.D., 5 μm) protected with a  Octadecylsilane (ODS) guard column (10 × 4.6 mm I.D., 5 μm). The mobile phase was aqueous solution (A) (containing 0.40% aqueous acetic acid) and acetonitrile (B). The conditions of the solvent gradient elution were 35-40% (B) in 0-15 min, 40-42% (B) in 15-18 min and 42-70% (B) in 18-30 min at a flow rate of 1.0 mL/min. Detection was conducted with wavelength of 270 nm at 30°C. Results: Good linearity relationships were found (r2 > 0.9955) over the investigated concentration range for bio-samples. Blood stasis was associated with significantly higher area under the concentration-time curve (AUC), the maximum plasma concentration (C max ) and biological half-life (t 1/2 ), lower total body clearance (CL) and apparent volume of distribution (Vd) of tanshinone IIA in plasma and higher AUC 0-t of tanshinone IIA in the analyzed tissues of rats treated with BHQ. Conclusion: Blood stasis could alter pharmacokinetics and tissue distribution of tanshinone IIA in BHQ.
  2 6,244 76
Strophanthus hispidus attenuates the Ischemia-Reperfusion induced myocardial Infarction and reduces mean arterial pressure in renal artery occlusion
Rohit Gundamaraju, Ravi Chandra Vemuri, Rajeev K Singla, Rishya Manikam, A Ranga Rao, Shamala Devi Sekaran
August 2014, 10(39):557-562
DOI:10.4103/0973-1296.139782  PMID:25298674
Background: The myocardium is generally injured in the case of reperfusion injury and arterial damage is caused by hypertension. In reference to these statements, the present study was focused. Cardiac glycosides were said to have protective effects against myocardial infarction and hypertension. Strophanthus hispidus was thus incorporated in the study. Objective: The prime objective of the study was to investigate the protective effects of Strophanthus hispidus against ischemia-reperfusion myocardial Infarction and renal artery occluded hypertension in rats. Materials and Methods: The animal model adopted was surgically-induced myocardial ischemia, performed by means of left anterior descending coronary artery occlusion (LAD) for 30 min followed by reperfusion for another 4 h. Infarct size was assessed by using the staining agent TTC (2,3,5-triphenyl tetrazolium chloride). Hypertension was induced by clamping the renal artery with renal bulldog clamp for 4 h. Results: The study was fruitful by the effect of Strophanthus hispidus on infarction size, which got reduced to 27.2 ± 0.5and 20.0 ± 0.2 by 500 mg/Kg and 1000 mg/Kg ethanolic extracts which was remarkably significant when compared with that of the control group 52.8 ± 4.6. The plant extract did reduce heart rate at various time intervals. There was also a protective effect in the case of mean arterial blood pressure were the 500 mg/Kg and 1000 mg/Kg of the plant extract did reduce the hypertension after 60 minutes was 60.0 ± 4.80 and 50.50 ± 6.80. Conclusion: The results suggest that 500 mg/Kg and 100 mg/Kg ethanolic extract of Strophanthus hispidus was found to possess significant cardiac protective and anti-hypertensive activity.
  2 2,259 151
Euphorbia hirta accelerates fibroblast proliferation and Smad-mediated collagen production in rat excision wound
Aadesh Upadhyay, Pronobesh Chattopadhyay, Danswrang Goyary, Papiya M Mazumder, Vijay Veer
August 2014, 10(39):534-542
DOI:10.4103/0973-1296.139801  PMID:25298671
Background: Euphorbia hirta L. (Euphorbiaceae) is a traditional herbal medicine used for treatment of various diseases. Objective: E. hirta was investigated for in vitro/in vivo wound healing activity using human dermal fibroblast cell line and Wistar rats. Materials and Methods: Petroleum ether, chloroform, methanol and water successive extracts of E. hirta leaves were evaluated for antioxidant, antimicrobial and fibroblast proliferation activities. Among different extracts, the promising methanol extract was screened for wound healing activity in Wistar rats, using gentamicin sulfate (0.01% w/w) as a reference. Wound contraction, hydroxyproline content and the protein expression of COL3A1, bFGF, Smad-2,-3,-4 and -7 were measured. Results: The E. hirta methanol extract showed a potent antimicrobial (MIC 0.250 mg/ml against Escherichia coli and Klebsiella pneumoniae, both), antioxidant activities (IC 50 = 10.57 μg/ml , 2,2-diphenyl-1-picrylhydrazyl; 850.23 μg/ml , superoxide-anion radical scavenging activity and 23.63 mg gallic acid equivalent per gram extract) with significant fibroblast proliferating activity (112% at 12.5 μg/ml) as compared to other extracts. In vivo study also supported the wound healing potential of methanol extract, as evidenced by faster wound contraction, higher hydroxyproline (4.240 mg/100 mg tissue) and improved histopathology of granulation tissue as compared to control groups and gentamicin sulfate-treated ones. Western blot also revealed a significantly altered expression of Smad-mediated proteins resulting in collagen production. Conclusion: The study suggested that E. hirta accelerates the wound healing by augmenting the fibroblast proliferation and Smad-mediated collagen production in wound tissue.
  2 3,531 114
So-Cheong-Ryong-Tang, a herbal medicine, modulates inflammatory cell infiltration and prevents airway remodeling via regulation of interleukin-17 and GM-CSF in allergic asthma in mice
Hyung-Woo Kim, Chi-Yeon Lim, Bu-Yeo Kim, Su-In Cho
August 2014, 10(39):506-511
DOI:10.4103/0973-1296.139784  PMID:25298667
Background: So-Cheong-Ryong-Tang (SCRT), herbal medicine, has been used for the control of respiratory disease in East Asian countries. However, its therapeutic mechanisms, especially an inhibitory effect on inflammatory cell infiltration and airway remodeling in allergic asthma are unclear. Objective: The present study investigated the mechanism of antiasthmatic effects of SCRT in allergic asthma in mice. Materials and Methods: We investigated the influence of SCRT on levels of interleukin-17 (IL-17), granulocyte/macrophage colony-stimulating factor (GM-CSF), IL-4, and interferon gamma (IFN-γ) in bronchoalveolar lavage fluid (BALF), ovalbumin (OVA)-specific IgE in serum, and histopathological changes in allergen-induced asthma. Results: So-Cheong-Ryong-Tang decreased levels of IL-17 and GM-CSF in BALF. IL-4, a Th2-driven cytokine, was also decreased by SCRT, but IFN-γ, a Th1-driven cytokine, was not changed. Levels of OVA-specific IgE in serum were also decreased by SCRT. With SCRT treatment, histopathological findings showed reduced tendency of inflammatory cell infiltration, and prevention from airway remodeling such as epithelial hyperplasia. Conclusion: In this study, we firstly demonstrated that regulation of IL-17 and GM-CSF production may be one of the mechanism contributed to a reduction of inflammatory cell infiltration and prevention from airway remodeling.
  2 2,416 93
Effect of whole plant of Rostellularia diffusa Willd. on experimental stress in mice
Mandipati Nagasirisha, TS Mohamed Saleem
August 2014, 10(39):614-621
DOI:10.4103/0973-1296.139799  PMID:25298682
Background: Rostellularia diffusa is an unexplored medicinal plant used as brain tonic in traditional medicine system. Objective: This study was designed to investigate the antioxidant and anti-stress potential of R. diffusa by experimental animal models. Materials and Methods: The extracts of R. diffusa were subjected to preliminary phytochemical screening and high performance thin layer chromatography (HPTLC) finger printing analysis. The antioxidant potential of the extracts was found by different in vitro models. The anti-stress activity was investigated by using acetic acid induced writhing test, swimming endurance test, and restraint stress in experimental mice. Serum parameters such as glucose, triglyceride and cholesterol, oxidative stress parameter thiobarbituric acid reactive substance, antioxidant parameters such as reduced glutathione, superoxide dismutase and catalase and organ weights were evaluated after restraint stress in mice. Diazepam was used as reference standard to compare the anti-stress activity of plant extract. Results: High performance thin layer chromatography finger printing analysis revealed the presence of flavone compounds in both extracts. The extracts also showed good antioxidant property in different in vitro antioxidant models. Administration of extracts of R. diffusa decreased the number of wriths and immobility time when compared with control group in acetic acid-induced writhing test and swimming endurance test respectively in experimental mice. They also suppressed the restraint stress-induced alterations in serum parameters, oxidative stress, and antioxidant parameters in brain and also restored the organ weights in normal level. Conclusion: From these results, it has been concluded that the potential anti-stress activity of R. diffusa is through its adaptogenic and antioxidant properties.
  2 2,399 70
Bioassay-guided isolation, identification and molecular ligand-target insight of lipoxygenase inhibitors from leaves of Anisomeles malabarica R.Br.
A Sudha, P Srinivasan
August 2014, 10(39):596-605
DOI:10.4103/0973-1296.139795  PMID:25298680
Background: Anisomeles malabarica R. Br. (Lamiaceae) is extensively used in traditional medicine in major parts of India for several medicinal purposes, including their use in rheumatism. Materials and Methods: The air-dried leaves of A. malabarica were extracted with ethanol, defatted with n-hexane and then successively partitioned into chloroform and n-butanol fractions. Bioassay-guided fractionation and purification of chloroform fraction from A. malabarica lead to the isolation of lipoxygenase (LOX) inhibitors. The structures of isolated compounds were elucidated by ultraviolet, infrared, 1 H nuclear magnetic resonance (NMR), 13 C NMR and mass spectrometry spectroscopic techniques and assessed further by in vitro soybean lipoxygenase (sLOX) assay. In addition, the enzyme type inhibition was evaluated through molecular docking technique as a part of computational study. Results: The bioactive compounds 3, 4 dihydroxy benzoic acid (1) and 4', 5, 7-trihydroxyflavone (2) were isolated from chloroform fraction of A. malabarica, whose bioactivity was observed to be dose-dependent compared to n-butanol fraction. Among the compounds, 3, 4 dihydroxy benzoic acid showed significant sLOX inhibitory activity with 74.04% ±2.6% followed by 4', 5, 7-trihydroxyflavone (34.68% ±1.9%). The computational analysis of compounds showed their molecular interaction with important amino acid residues and nonheme iron atom in the catalytic site of LOX by enlightening their potential binding mode at molecular level. Conclusions: The LOX inhibitory constituents were identified from A. malabarica by means of bioassay-guided fractionation process. The results derived from in vitro and computational experiments confirm the potential of the isolated compounds and provide additional evidence for its traditional use in inflammatory disorders.
  2 3,713 151
Anti-cancer effects of Kochia scoparia fruit in human breast cancer cells
Hye-Yeon Han, Hyungwoo Kim, Yong Hae Son, Guemsan Lee, Sung-Hee Jeong, Mi Heon Ryu
August 2014, 10(39):661-667
DOI:10.4103/0973-1296.139812  PMID:25298688
Background: The fruit of Kochia scoparia Scharder is widely used as a medicinal ingredient for the treatment of dysuria and skin diseases in China, Japan and Korea. Especially, K. scoparia had been used for breast masses and chest and flank pain. Objective: To investigate the anti-cancer effect of K. scoparia on breast cancer. Materials and Methods: We investigated the anti-cancer effects of K. scoparia, methanol extract (MEKS) in vitro. We examined the effects of MEKS on the proliferation rate, cell cycle arrest, reactive oxygen species (ROS) generation and activation of apoptosis-associated proteins in MDA-MB-231, human breast cancer cells. Results: MTT assay results demonstrated that MEKS decreased the proliferation rates of MDA-MB-231 cells in a dose-dependent manner with an IC 50 value of 36.2 μg/ml. MEKS at 25 μg/ml significantly increased the sub-G1 DNA contents of MDA-MB-231 cells to 44.7%, versus untreated cells. In addition, MEKS induced apoptosis by increasing the levels of apoptosis-associated proteins such as cleaved caspase 3, cleaved caspase 8, cleaved caspase 9 and cleaved Poly (ADP-ribose) polymerase (PARP). Conclusion: These results suggest that MEKS inhibits cell proliferation and induces apoptosis in breast cancer cells and that MEKS may have potential chemotherapeutic value for the treatment of human breast cancer.
  2 2,563 178
Antinociceptive effect of Encholirium spectabile: A Bromeliaceae from the Brazilian caatinga biome
Sarah Raquel Gomes de Lima-Saraiva, Juliane Cabral Silva, Carla Rodrigues Cardoso Branco, Alexsandro Branco, Elba Lúcia Cavalcanti Amorim, Jackson Roberto Guedes da Silva Almeida
August 2014, 10(39):655-660
DOI:10.4103/0973-1296.139817  PMID:25298687
Background: Encholirium spectabile is a species found in outcrops rocky throughout the Brazilian Caatinga. Objective: This study was carried out to evaluate the antinociceptive effects of ethanolic extract of the leaves from E. spectabile (Es-EtOH) in mice using chemical and thermal models of nociception. Material and Methods: HPLC was used to determine the fingerprint chromatogram. The Es-EtOH was examined for its antinociceptive activity at the doses of 100, 200 and 400 mg/kg intraperitoneal (i.p.). The evaluation of antinociceptive activity was carried out by the acetic acid-induced writhing, formalin and hot plate tests in mice. Rota-rod test was used for the evaluation of motor coordination. Results: In the acetic acid-induced writhing test, the Es-EtOH (100, 200 and 400 mg/kg, i.p.) reduced the number of writhings by 68.59, 79.33 and 65.28%, respectively. Additionally, Es-EtOH (100, 200 and 400 mg/kg, i.p.) decreased by 34.14, 52.61 and 60.97% the paw licking time in the first phase, as well as 89.56, 79.90 and 96.71% in the second phase of the formalin test, respectively. Es-EtOH also showed effect in the hot plate test, since increased the latency time at dose of 100 mg/kg after 60 minutes. In addition, Es-EtOH did not impair motor coordination. The presence of phenolic compounds in the extract was confirmed using HPLC. These results indicate that Es-EtOH has antinociceptive activity, probably of peripheral origin. The mechanism involved is not completely understood but, at least in part there is the participation of opioid receptors.
  1 2,018 67
The Pavonia xanthogloea (Ekman, Malvaceae): Phenolic compounds quantification, anti-oxidant and cytotoxic effect on human lymphocytes cells
Clarice Pinheiro Mostardeiro, Marco Aurélio Mostardeiro, Ademir Farias Morel, Raul Moreira Oliveira, Alencar Kolinski Machado, Pauline Ledur, Francine Carla Cadoná, Ubiratan Flores da Silva, Ivana Beatrice Mânica da Cruz
August 2014, 10(39):630-638
DOI:10.4103/0973-1296.139804  PMID:25298684
Introduction: Pavonia xanthogloea is traditionally used as an antimicrobial and anti-tumour medicine in Southern Brazilian region. However, investigations about this species are still incipient. Hypothesis Tested: The study postulated that P. xanthologea specie present some phenolic compound and present some biological properties as anti-oxidant and cytoprotective effect against oxidative stress. Materials and Methods: The content of eight phenolic molecules in the crude ethanolic extract of the aerial part of P. xanthogloea and its five fractions (hexane, dichloromethane, ethyl-acetate, n-butanol, and water) was determined by heterotrophic plate count method. The anti-oxidant capacity of the extract and the fractions was determined by 1,1-diphenyl-2-picryl-hydrazyl assay. The potential anti-oxidant and cytoprotective effect was also analyzed in human lymphocyte culture treated with extract/fractions at different concentrations with and without oxidative stress generated by hydrogen peroxide (H 2 O 2 ) and sodium nitroprusside (SNP) exposition. Results: Tiliroside was the molecule detected in all extract. Water and ethyl-acetate fractions showed the highest radical-scavenging activity. The crude extract, hexane, water, and n-butanol reversed the higher reactive oxygen specie levels generated by H 2 O 2 and SNP to levels similar to those observed in the control group. In addition, crude extract, hexane, ethyl-acetate and n-butanol did not caused cytotoxicity, whereas water fraction was cytotoxic at higher concentration tested here (300 μg/mL). The cytotoxicity reversion caused by SNP exposition was concentration-dependent of the extract and fractions. However, dichloromethane fraction increased cell mortality in all concentrations investigated and was not able to decrease cell death in the lymphocytes exposed to SNP. Conclusion: The results suggest potential medicine use of this species.
  1 2,143 72
Efficacy evaluation of Bauhinia variegata L. stem bark powder as adjunct therapy in chronic Staphylococcus aureus mastitis in goat
Jeevan Ranjan Dash, Tapas Kumar Sar, Indranil Samanta, Subodh Pal, Madhuchhanda Khan, Nimai Chand Patra, Uttam Sarkar, Asit Kumar Maji, Tapan Kumar Mandal
August 2014, 10(39):512-518
DOI:10.4103/0973-1296.139786  PMID:25298668
Objective: The objective was to study the effect of Bauhinia variegata L. stem bark powder as adjunct therapy in chronic Staphylococcus aureus mastitis in goat. Materials and Methods: Mastitis was induced by intracisternal inoculation of coagulase positive S. aureus (J638) at the concentration of 2000 colony forming units. Group I animals were treated with repeated dose of ceftriaxone at 20 mg/kg intravenously, and Group II animals were treated with once daily oral administration of B. variegata L. stem bark powder at 6 g/kg for 7 days followed by maintenance dose at 3 g/kg for next 7 days along with repeated dose of the antibiotic at 20 mg/kg intravenously at 4 days interval. Results: No significant improvement in the clinical condition of the udder was noticed in the group treated with repeated dose of ceftriaxone alone. However, in the group treated with B. variegata L. stem bark powder along with repeated dose of ceftriaxone, no S. aureus colony was seen at 96 h and onwards in milk samples with a marked decrease in somatic cell count and milk alkaline phosphatase activity and increased lactoperoxidase activity. Further, plasma and milk concentration of ceftriaxone/ceftizoxime was increased, which indicated antibacterial, bioenhancing and antiinflammatory properties of the bark powder. The Group II animals also exhibited marked reduction in polymorphonuclear cells and fibrous tissue indicating antifibrotic property of B. variegata L. Conclusion: B. variegata L. stem bark powder can be considered as an effective adjunct therapy to intravenous ceftriaxone in S. aureus chronic mastitis in goat.
  1 4,142 81
The effects of extraction method on recovery rutin from Calendula officinalis L. (Asteraceae)
Frederico Severino Martins, Edemilson Cardoso da Conceição, Elane Sousa Bandeira, José Carréra Silva, Roseane Maria Ribeiro Costa
August 2014, 10(39):569-573
DOI:10.4103/0973-1296.139785  PMID:25298676
Background: Calendula officinalis L. (Asteraceae) is a Mediterranean specie, but in Europe and America it is cultivated for ornamental or medicinal purposes. This species is widely used for presenting activities, antiinflammatory antibacterial and antioxidant. However the therapeutic action is linked to the amount of assets of the extracted raw material. The extraction method of bioactive compounds is an important step in the manufacturing of herbal medicines, because secondary metabolites with therapeutic potential are usually found in small quantities in plant materials. Objective: Due the medical and commercial importance of C. officinalis, this study aimed to evaluate the impact of the extraction method on the quality of herbal extract and optimize the extraction of rutin from C. officinalis. Materials and Methods : The extraction of rutin was performed by ultrasound and shaker and the optimized conditions were determined by response surface methodology. Results: The results of ultrasound extraction assisted (UEA) and maceration dynamic (MD) showed that rutin yield ranged from 0.218 to 2.28% (w/w) when extract by ultrasound and 0.1-1.44% by MD. The optimal extraction condition for rutin (2.48% to UEA or 1.46% to MD) from C. officinalis by UEA or MD were a 19-22 min extraction, ethanol: water ratio of 35-40% and 0.05-0.056 mg/mL to raw material: solvent ratio. Conclusion: The UEA is more efficient to extraction rutin.
  1 2,537 140
Chemical composition and hepatotoxic effect of Geranium schiedeanum in a thioacetamide-induced liver injury model
Juan Gayosso-De-Lucio, Mirandeli Bautista, C Velazquez-González, M De la O Arciniega, JA Morales-González, Juana Benedí
August 2014, 10(39):574-580
DOI:10.4103/0973-1296.139788  PMID:25298677
One of the major components of some geraniums is geraniin, described by its discoverer as crystallizable tannin, well known as an excellent antioxidant, and also found in fruits such as pomegranate. Recently, natural antioxidants have attracted great attention from consumers over the world due to their lower toxicity than synthetics. But geraniin is not a stable compound, and also is difficult to obtain, that is why in the present study we obtained acetonylgeraniin from Geranium schideanum (Gs), a stable acetone condensate of geraniin. In the present study the effect of Gs acetone-water extract was studied in reference to postnecrotic liver regeneration induced by thioacetamide (TA) in rats. Two months male rats were pretreated with daily dose of Gs extract for 4 days (300 mg/kg) and the last day also were intraperitoneally injected with TA (6.6 mmol/kg). Samples of blood were obtained from rats at 0, 24, 48, 72 and 96 h following TA intoxication. The pre-treatment with the crude extract in the model of thioacetamide-induced hepatotoxicity in rats decreased and delayed liver injury by 66% at 24 h. This result suggests that Gs extract may be used as an alternative for reduction of liver damage. On the other hand, acute toxicity study revealed that the LD 50 value of the Gs extract is more than the dose 5000 mg/kg in rats, according to the Lorke method.
  1 2,661 103
Direct proof by 13C-nuclear magnetic resonance of semi-purified extract and isolation of ent -Catechin from leaves of Eucalyptus cinerea
Sayonara Mendes Silva, Simone Yae Abe, Fernanda Giacomini Bueno, Norberto Peporine Lopes, João Carlos Palazzo de Mello, Tomoe Nakashima
July-September 2014, 10(39):191-194
DOI:10.4103/0973-1296.137355  PMID:25210302
Background: Eucalyptus cinerea F. Muell. ex Benth. is native to Australia and acclimatized to Southern Brazil. Its aromatic leaves are used for ornamental purposes and have great potential for essential oil production, although reports of its use in folk medicine are few. Objective: This study evaluated the composition of E. cinerea leaves using the solid state 13 C-nuclear magnetic resonance (NMR) and isolation of the compound from the semipurified extract (SE). Materials and Methods: The SE of E. cinerea leaves was evaluated in the solid state by 13 C-NMR spectrum, and the SE was chromatographed on a Sephadex LH-20 column, followed by high-speed counter-current chromatography to isolate the compound. The SE was analyzed by 13 C-NMR and matrix-assisted laser desorption/ionization-time-of-flight spectra. Results: Flavan-3-ol units were present, suggesting the presence of proanthocyanidins as well as a gallic acid unit. The uncommon ent-catechin was isolated. Conclusion: The presence of ent-catechin is reported for the first time in this genus and species.
  1 4,360 79
Preparation and pharmacokinetics in beagle dogs of ganershu sustained-release pellets
Jin-huo Pan, Jian-chun Wang, Zhi-tao Jiang, Ting Zhang, Shao-bo Ge, Ye-xia Zhang, Xin Jin, Guo-jun Yan
July-September 2014, 10(39):217-226
DOI:10.4103/0973-1296.137360  PMID:25210307
Background: The active ingredients of Ganershu compound recipe, which are effective for hepatitis treatment in liver protection and transaminase reduction. However, the active ingredients of Ganershu compound recipe are poor absorption, which conduct it has a low oral bioavailability. Objective: We prepared Ganershu sustained-release pellets (GSPs) by fluidized-bed on central composite design-response surface methodology and increase its bioavailability in beagle dogs. Materials and Methods: In this study, GSPs were successfully prepared. The Drug-loaded pellets and sustained-release coated were carried out in fluidized-bed machine. GSP was optimized for fitting release, roundness, and the overall desirability by central composite design-response surface methodology. Results: To optimize cumulative release profile, the outermost ethyl cellulose coating layer and the hydroxypropyl methyl cellulose (HPMC) swelling layer were employed, which were respectively given coating levels in terms of weight gain of 22% and 6%, the concentration of HPMC is 4.5% (g/ml). The pharmacokinetics of Ganershu normal pellets (GNPs) and GSP was studied in beagle dogs after oral administration. The naringenin as an index, the area under the curve 0-∞ of naringenin in GSP was 1.38 times greater than that of GNP. Meanwhile, Tmax of GSP was prolonged for about 74%. Conclusion: This study can clearly indicate that we enhanced the oral bioavailability of Ganershu by preparing the GSP, which had the sustained dissolution and improved the potential of it for clinical application.
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New steroidal saponin from Antigonon leptopus Hook. and Arn.
Maria Karmella L Apaya, Christine L Chichioco-Hernandez
August 2014, 10(39):501-505
DOI:10.4103/0973-1296.139781  PMID:25298666
Background: Antigonon leptopus Hook. and Arn., Polygonaceae (cadena de amor), is a herbal remedy for pain and gout-like symptoms in the Philippines. The methanol extract of A. leptopus have shown strong inhibitory action against xanthine oxidase. Objective: To isolate and identify the compound responsible for the xanthine oxidase inhibitory action. Materials and Methods: A bioassay-guided isolation scheme using an in vitro assay for the inhibition of xanthine oxidase was employed. The structure was established using spectroscopic analysis and chemical methods. Results: The isolated compound was determined to be a noncompetitive inhibitor of xanthine with an IC50 of 1.79 μg/mL. Conclusion: The isolated compound may represent a new class of xanthine oxidase inhibitors.
  1 2,617 95
Ultra high performance liquid chromatography- ultraviolet-electrospray ionization-micrOTOF-Q II analysis of flavonoid fractions from Jatropha tanjorensis
Arun Kallur Purushothaman, Brindha Pemiah
August 2014, 10(39):472-479
DOI:10.4103/0973-1296.139776  PMID:25298662
Background: Jatropha tanjorensis (Euphorbiaceae) an exotic traditional plant unique to Thanjavur district of Southern India also commonly called as Catholic vegetable. It has been used traditionally in decoctions for treating various ailments and as a health tonic. Objective: The objective of the present work is to study a comprehensive characterization of methanolic extract fractions using ultra high performance liquid chromatography (UHPLC)+-electrospray ionization (ESI)-micrOTOF-Q II and correlate their bioactivities. Materials and Methods: Phytoconstituents from J. tanjorensis leaves were extracted with methanol (MeOH) followed by successive chromatography using linear gradient polar solvents system. All fractions obtained were evaluated for their chemical potential using micrOTOF-Q II techniques and identified key molecules were determined for their anticancer and anti-oxidant potential using in vitro methods. Results: Successive column chromatography of the MeOH residue yielded six fractions. Compounds such as such as C-glycosylflavones (mono-C-, di-C-), O, C-diglycosylflavones and aglycones were identified for the first time in this plant using UHPLC-ultraviolet-micrOTOF-Q II ESI and a correlation with their anticancer using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay on Ehrlich ascites cells (EAC) and antioxidant activities using 2,2-diphenyl-1-picrylhydrazyl and lipid peroxidation were studied; fraction D extract exhibited the strongest activity against cancer cell. Conclusions: LC-mass spectrometry has been successfully applied for a quick separation and identification of the major phytoconstituents. All fractions have shown potent antioxidative activity as compared to standard antioxidant 3,5-di-tert-butyl-4-hydroxytoluene. EAC cell-based cytotoxicity assay also revealed encouraging results. The antioxidant and anticancer activity determined in the present work can be attributed to the presence of flavonoids and flavone glycosides. Present work provides the first scientific report on phytoconstituents of J. tanjorensis and its ethnopharmacological significance.
  1 2,415 78
Simultaneous determination of four neuroprotective compounds of Tilia amurensis by high performance liquid chromatography coupled with diode array detector
Bohyung Lee, Jin Bae Weon, Bo-Ra Yun, Jiwoo Lee, Min Rye Eom, Choong Je Ma
July-September 2014, 10(39):195-199
DOI:10.4103/0973-1296.137353  PMID:25210303
Background: Tilia amurensis consists of various compounds, such as flavonoids and terpenoids. Objective: A simple and reliable high performance liquid chromatography (HPLC) coupled with the diode array detector (DAD) method has been established for simultaneous determination of epicatechin, nudiposide, lyoniside, and scopoletin isolated from Tilia amurensis. Materials and Methods: Optimum separations were obtained with a SHISEIDO C 18 column by gradient elution, with 0.1% Trifluoroacetic acid (TFA) water-methanol as the mobile phase. The gradient elution system was completed within 40 minutes. The flow rate and detection wavelength were 1 mL/minute, 205 nm, 250 nm, and 280 nm, respectively. Results: Validation of the analytical method was evaluated by linearity, precision, and the accuracy test. The calibration curve was linear over the established range with R 2 > 0.997. The limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.01-15.20 μg/mL and 0.03-46.06 μg/mL. The method exhibited an intraday and interday precision range of 96.25-105.66% and 93.52-109.92%, respectively (RSD <2.80%). The recoveries and relative standard deviation (RSD) of the four compounds in Tilia amurensis were in the range of 90.42-104.84% and 0.2-2.58%. Conclusion: This developed method was accurate and reliable for the quality evaluation of the four compounds isolated from Tilia amurensis.
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