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   2015| Jul-Sep  | Volume 11 | Issue 43  
    Online since July 10, 2015

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Anti-aging effect of polysaccharide from Bletilla striata on nematode Caenorhabditis elegans
Yusi Zhang, Ting Lv, Min Li, Ting Xue, Hui Liu, Weiming Zhang, Xiaoyu Ding, Ziheng Zhuang
Jul-Sep 2015, 11(43):449-454
DOI:10.4103/0973-1296.160447  PMID:26246718
Background: Polysaccharide isolated from Bletilla striata, a well known traditional Chinese medicine (Bletilla striata polysaccharide [BSP]) has been found to play important roles in endothelial cells proliferation, inducible nitric oxide stimulation, wound healing acceleration and other processes. Recent studies found that B. striata has anti oxidative properties, however, potential anti aging effects of BSP in whole organisms has not been characterized. Objective: To investigate whether BSP has anti aging effects on Caenorhabditis elegans. Materials and Methods: After treatment with BSP, the lifespan, locomotion ability, and stress resistance of C. elegans was determined. To provide insight into the underlying mechanism for the anti aging effect of BSP, we measured its effect on bacterial growth, brood size of C. elegans, and the insulin/insulin like growth factor (IGF) signaling pathway. Results: After BSP treatment, the lifespan of C. elegans was extended, and its locomotion ability and stress resistance were increased. BSP was found to have no effect on bacterial growth or on reproduction of C. elegans, However, mRNA levels of age-1 and hcf-1 were reduced after BSP treatment. Additionally, we observed that BSP did not extend the lifespan of daf 16 mutant animals. Conclusion: BSP produces an anti aging effect on C. elegans through the insulin/IGF signaling pathway and holds promise for future development as a functional food.
  24 5,479 280
Luteolin is a bioflavonoid that attenuates adipocyte-derived inflammatory responses via suppression of nuclear factor-κB/mitogen-activated protein kinases pathway
Sarmila Nepali, Ji-Seon Son, Barun Poudel, Ji-Hyun Lee, Young-Mi Lee, Dae-Ki Kim
Jul-Sep 2015, 11(43):627-635
DOI:10.4103/0973-1296.160470  PMID:26246742
Background: Inflammation of adipocytes has been a therapeutic target for treatment of obesity and metabolic disorders which cause insulin resistance and hence lead to type II diabetes. Luteolin is a bioflavonoid with many beneficial properties such as antioxidant, antiproliferative, and anti cancer. Objectives: To elucidate the potential anti inflammatory response and the underlying mechanism of luteolin in 3T3 L1 adipocytes. Materials and Methods: We stimulated 3T3 L1 adipocytes with the mixture of tumor necrosis factor a, lipopolysaccharide, and interferon g (TLI) in the presence or absence of luteolin. We performed Griess’ method for nitric oxide (NO) production and measure mRNA and protein expressions by real time polymerase chain reaction and western blotting, respectively. Results: Luteolin opposed the stimulation of inducible nitric oxide synthase and NO production by simultaneous treatment of adipocytes with TLI. Furthermore, it reduced the pro inflammatory genes such as cyclooxygenase 2, interleukin 6, resistin, and monocyte chemoattractant protein 1. Furthermore, luteolin improved the insulin sensitivity by enhancing the expression of insulin receptor substrates (IRS1/2) and glucose transporter 4 via phosphatidylinositol 3K signaling pathway. This inhibition was associated with suppression of IkB a degradation and subsequent inhibition of nuclear factor κB (NF κB) p65 translocation to the nucleus. In addition, luteolin blocked the phosphorylation of ERK1/2, c Jun N terminal Kinases and also p38 mitogen activated protein kinases (MAPKs). Conclusions: These results illustrate that luteolin attenuates inflammatory responses in the adipocytes through suppression of NF κB and MAPKs activation, and also improves insulin sensitivity in 3T3 L1 cells, suggesting that luteolin may represent a therapeutic agent to prevent obesity associated inflammation and insulin resistance.
  18 3,791 142
Metabolic fingerprinting to understand therapeutic effects and mechanisms of silybin on acute liver damage in rat
Qun Liang, Cong Wang, Binbing Li, Ai-hua Zhang
Jul-Sep 2015, 11(43):586-593
DOI:10.4103/0973-1296.160469  PMID:26246736
Background: Metabolic fingerprinting is a rapid and noninvasive analysis, representing a powerful approach for the characterization of phenotypes and the distinction of specific metabolic states due to environmental alterations. It has become a valuable analytical approach for the characterization of phenotypes and is the rapidly evolving field of the comprehensive measurement of ideally all endogenous metabolites in bio-samples. Silybin has displayed bright prospects in the prevention and therapy of liver injury, and we had conducted a preliminary exploration on the molecular mechanism of the hepatoprotective effects of silybin. Because the knowledge on the metabolic responses of an acute liver damage rat to the silybin is still scarce, metabolic fi ngerprinting can provide relevant information on the intrinsic metabolic adjustments. Materials and Methods: Here, the physiological and metabolic changes in the acute liver damage rat were investigated by performing a metabolic analysis. The phenotypic response was assessed by liquid chromatography/mass spectrometry (LC/MS) combined with pattern recognition approaches such as principal components analysis and partial least squares projection to supervised latent structures and discriminant analysis. Multivariate analysis of the data showed trends in scores plots that were related to the concentration of the silybin. Results: Results indicate 10 ions (7 upregulated and 3 downregulated) as differentiating metabolites. Key observations include perturbations of metabolic pathways linked to glutathione metabolism, tryptophan metabolism, cysteine and methionine metabolism, etc., Overall, this investigation illustrates the power of the LC/MS combined with the pattern recognition methods that can engender new insights into silybin affecting on metabolism pathways of an acute liver damage rat. Conclusion: The present study demonstrates that the combination of metabolic fi ngerprinting with appropriate chemometric analysis is a valuable approach for studying cellular responses to silybin drug and can provide additional insight into the mechanisms.
  13 3,937 371
Chemical constituents and antioxidant activity of the essential oil from leaves of Annona vepretorum Mart. (Annonaceae)
Camila de Souza Ara˙jo, Ana Paula de Oliveira, Rafaely Nascimento Lima, Péricles Barreto Alves, Tâmara Coimbra Diniz, Jackson Roberto Guedes da Silva Almeida
Jul-Sep 2015, 11(43):615-618
DOI:10.4103/0973-1296.160462  PMID:26246740
Background: Annona vepretorum (AV) is a native tree from Caatinga biome (semiarid region of Brazil) popularly known as “araticum” and “pinha da Caatinga.” Objective: This study was carried out to evaluate the chemical constituents and antioxidant activity (AA) of the essential oil from the leaves from AV (EO Av) collected in Petrolina, Pernambuco, Brazil. Materials and Methods: Fresh leaves of AV were cut into pieces, and subjected to distillation for 2 h in a clevenger type apparatus. Gas chromatograph (GC) analyses were performed using a mass spectrometry/flame ionization detector. The identification of the constituents was assigned on the basis of comparison of their relative retention indices. The antioxidant ability of the EO was investigated through two in vitro models such as radical scavenging activity using 2,2 diphenyl 1 picrylhydrazyl method and β-carotene linoleate model system. The positive controls (ascorbic acid, butylated hydroxyanisole and butylated hydroxytoluene) were those using the standard solutions. Assays were carried out in triplicate. Results: The oil showed a total of 21 components, and 17 were identified, representing 93.9% of the crude EO. Spathulenol (43.7%), limonene (20.5%), caryophyllene oxide (8.1%) and a pinene (5.5%) were found to be the major individual constituents. Spathulenol and caryophyllene oxide could be considered chemotaxonomic markers of these genera. The EO demonstrated weak AA.
  13 1,753 22
A direct protein kinase B-targeted anti inflammatory activity of cordycepin from artificially cultured fruit body of Cordyceps militaris
Ju Young Yoon, Ji Hye Kim, Kwang-Soo Baek, Geum Soog Kim, Seung Eun Lee, Dae Young Lee, Je Hun Choi, Seung Yu Kim, Hyun Bong Park, Gi-Ho Sung, Kang Ro Lee, Jae Youl Cho, Hyung Jun Noh
Jul-Sep 2015, 11(43):477-485
DOI:10.4103/0973-1296.160454  PMID:26246722
Background: Cordyceps militaris is one of well-known medicinal mushrooms with anti-inflammatory, anti-cancer, anti-diabetic, and anti-obesity activities. Objective: The objective of the following study is to isolate chemical components from the ethanol extract (Cm-EE) from Cordyceps militaris and to evaluate their anti-inflammatory activities. Materials and Methods: Column chromatographic separation was performed and anti-inflammatory roles of these compounds were also examined by using NO production and protein kinase B (AKT) activity assays. Results: From Cm-EE, 13 constituents, including trehalose (1), cordycepin (2), 6-hydroxyethyladenosine (3), nicotinic amide (4), butyric acid (5), β-dimorphecolic acid (6), α-dimorphecolic acid (7), palmitic acid (8), linoleic acid (9), cordycepeptide A (10), 4-(2-hydroxy-3-((9E,12E)-octadeca-9,12-dienoyloxy)propoxy)-2-(trimethylammonio)butanoate (11), 4-(2-hydroxy-3-(palmitoyloxy)propoxy)-2-(trimethylammonio)butanoate (12), and linoleic acid methyl ester (13) were isolated. Of these components, compound 2 displayed a significant inhibitory effect on NO production in lipopolysaccharide (LPS)-activated RAW264.7 cells. Furthermore, this compound strongly and directly suppressed the kinase activity of AKT, an essential signalling enzyme in LPS-induced NO production, by interacting with its ATP binding site. Conclusion: C. militaris could have anti-inflammatory activity mediated by cordycepin-induced suppression of AKT.
  13 4,127 195
Profiling of components of rhizoma et radix polygoni cuspidati by high-performance liquid chromatography with ultraviolet diode-array detector and ion trap/time-of-flight mass spectrometric detection
Jinfeng Fu, Min Wang, Huimin Guo, Yuan Tian, Zunjian Zhang, Rui Song
Jul-Sep 2015, 11(43):486-501
DOI:10.4103/0973-1296.160455  PMID:26246723
Introduction: Rhizoma et Radix Polygoni Cuspidati (Huzhang in Chinese, HZ) is a traditional medicinal plant in China. Many of the components of HZ have been proved to be bioactive while it is difficult to conduct a comprehensive chemical profiling of HZ as a consequence of the absence of efficient separation system and sensitive detective means. We developed a simple and effective method for comprehensive characterization of constituents in HZ. Objective: To develop a simple and effective method to characterize the components in HZ and provide useful information for subsequent metabolic studies of HZ. Materials and Methods: The components in HZ aqueous extract were characterized by using high performance liquid chromatography with UV diode-array detector (HPLC-DAD) and ion trap/time-of-flight mass spectrometric detection (HPLC-IT/TOF). Stilbenes, anthraquinones, gallates and tannins, naphthalenes and some other compounds were identified and confirmed by diagnostic fragment ions with accurate mass measurements, characteristic fragmentation pathways and relevant published literatures. Results: Among the 238 constituents detected in HZ, a total number of 74 constituents were identified unambiguously or tentatively, including 29 compounds reported for the first time in HZ. Conclusion: The identification and structure elucidation of these chemicals provided essential data for quality control and further in vivo metabolic studies of HZ.Key words: Polygonum cuspidatum, HPLC-DAD, HPLC-IT/TOF, qualitative analysis.
  12 4,323 223
Phytochemical screening, antioxidant and antibacterial activities of extracts prepared from different tissues of Schinus terebinthifolius Raddi that occurs in the coast of Bahia, Brazil
Cinara Oliveira D'Sousa' Costa, Paulo Roberto Ribeiro, Marta Bruno Loureiro, Rafael Conceição Simões, Renato Delmondez de Castro, Luzimar Gonzaga Fernandez
Jul-Sep 2015, 11(43):607-614
DOI:10.4103/0973-1296.160459  PMID:26246739
Background: Schinus terebinthifolius is widely used in traditional medicine by Brazilian quilombola and indigenous communities for treatment of several diseases. Extracts from different tissues are being used to produce creams to treat cervicitis and cervicovaginitis. However, most studies are limited to the assessment of the essential oils and extracts obtained from the leaves. Objective: The aim was to evaluate antioxidant and antibacterial activities, to assess the phytochemical profile and to quantify total phenolic compounds of various extracts prepared from S. terebinthifolius grown in the coast of Bahia, Brazil. Materials and Methods: Extracts were obtained by hot continuous extraction (soxhlet) and by maceration. Quantification of phenolic compounds was performed using the Folin Ciocalteu method and antioxidant properties were assessed by 2,2 diphenyl 1 picrylhydrazyl radical scavenging assay. Phytochemical screening was performed as described by in the literature and antibacterial activity against Enterococcus faecalis (ATCC 29212) was determined by the microdilution broth assay. Results: Extraction method greatly affected the metabolite profile of the extracts. Antioxidant activity varied between 21.92% and 85.76%, while total phenols ranged between 5.44 and 309.03 mg EAG/g of extract. Leaf extract obtained with soxhlet showed minimum inhibitory concentration (MIC) of 15.62 μg/mL, while stem extract obtained by maceration was able to inhibit the growth of E. faecalis at 62.5 μg/mL. Stem bark extracts showed a MIC of 500 μg/mL for both extraction methods, while no inhibition was observed for fruit extracts. Conclusion: In general, total phenolic content, antioxidant and antibacterial activities were higher in samples obtained by soxhlet. Our results provide important clues in order to identify alternative sources of bioactive compounds that can be used to develop new drugs.
  11 4,890 58
Flavonoids isolated from Lespedeza cuneata G. Don and their inhibitory effects on nitric oxide production in lipopolysaccharide-stimulated BV - 2 microglia cells
Guijae Yoo, Seon Ju Park, Taek Hwan Lee, Heejung Yang, Yoon-su Baek, Nanyoung Kim, Yoon Jae Kim, Seung Hyun Kim
Jul-Sep 2015, 11(43):651-656
DOI:10.4103/0973-1296.160466  PMID:26246745
Background: Lespedeza cuneata (Dum. Cours.) G. Don, a perennial legume native to Eastern Asia, has been used therapeutically in traditional Asian medicine to protect the function of liver, kidneys and lungs. However, its effect on inflammatory nitric oxide (NO) production and the active constituents have not yet been explored. Objective: In this study, we investigated the phytochemical constituents of L. cuneata and evaluated their effect on NO production using lipopolysaccharide (LPS) stimulated BV2 cells. Materials and Methods: The 80% methanol extract of the aerial part of L. cuneata were used for the isolation of flavonoids. The isolated compounds were elucidated by various spectroscopic methods including nuclear magnetic resonance and mass spectrometry spectrometry. To evaluate the effect on inflammatory NO production, LPS stimulated murine microglia BV 2 cells were used as a screening system. Results: Nine flavonoids were isolated from the aerial parts of L. cuneata. Among the isolated flavonoids, compounds 4, 5, 7 and 9 are reported from the genus Lespedeza for the first time. Moreover, compounds 1 and 6 showed significant inhibitory effects on NO production in LPS stimulated BV2 cells without cell toxicity. Conclusion: In this study, nine flavonoids were isolated from L. cuneata. Among the compounds, only 1 and 6, which have free hydroxyl groups at both C3 and C7 showed significant inhibitory activity on NO production in LPS stimulated BV2 cells. These results suggested L. cuneata and its flavonoid constituents as possible candidate for the treatment of various inflammatory diseases.
  10 3,265 117
In situ pressurized biphase acid hydrolysis, a promising approach to produce bioactive diosgenin from the tubers of Dioscorea Zingiberensis
Huan Yang, Hua-wu Yin, Xue-wei Wang, Zi-hao Li, Yu-ping Shen, Xiao-bin Jia
Jul-Sep 2015, 11(43):636-642
DOI:10.4103/0973-1296.160472  PMID:26246743
Background: The tubers of Dioscorea zingiberensis, is the most favorable plant material for the production of diosgenin, an important bioactive steroidal sapogenin and requisite precursor of cortin, contraceptive and sex hormone, which is the only desired product after steroidal saponins from the tubers are hydrolyzed. Objective: A novel technology, in situ pressurized biphase acid hydrolysis was constructed for the first time to simplify extraction process, increase extraction yield and decrease the consumption of mineral acids. Materials and Methods: The method developed in this study has been optimized and verified through orthogonal design for experiments, in which the effect and their significance of four factors including molarity of acid, temperature, extraction duration and sample quantity have been investigated. Then, the comparison was conducted among the newly developed method and other reported methods. The diosgenin was also isolated by column chromatography, followed by mass spectrometry and nuclear magnetic resonance analysis for structural confirmation. Results: It was found that temperature is the factor of the most influence and the highest extraction yield at 2.21% has been achieved while the hydrolysis was performed at 140°C for 1.5 h in 0.20M H2SO4 solution with petroleum ether under an uncontrolled pressurized condition. And, compared to the others, the increment in the extraction yield of new method was 20.8 ~ 74.0%, and the consumption of H2SO4 was reduced by 17 times at most. Conclusion: This method is a much cleaner and more efficient approach for extraction of diosgenin from the tubers, and is promising to be applied in pharmaceutical industry.
  9 3,416 136
Purification and characterization of an anticoagulant oligopeptide from Whitmania pigra Whitman
Xiaobei Zheng, Juan Li, Zhengwang Chen, Yimei Liu, Keli Chen
Jul-Sep 2015, 11(43):444-448
DOI:10.4103/0973-1296.160446  PMID:26246717
Background: Dried Whitmania pigra is used for the treatment of cardiovascular and cerebrovascular diseases in traditional Chinese medicine and hot water and alcohol extracts also have anticogulant activity. However, a lower molecular weight and more stable anticogulant is needed. Objective: The objective of the following study is to purify and characterize of an anticoagulant oligopeptide from Hirudo (Whitmania pigra Whitman). Materials and Methods: Gel filtration on Sephadex G 50, ion exchange on diethylaminoethyl cellulose, and semi prepared high performance liquid chromatography were used to purify Hirudo. Automated coagulation analyzer was used for evaluating anticoagulant activity. Molecular weight was measured by Matrix assisted laser desorption ionization time of flight mass spectrometry. Amino acid sequence of the oligopeptide was measured by amino acid sequence analyzer. Results: A new anticoagulant, named whitide, isolated from Hirudo was purified, with a molecular weight 1997.1 Da. Amino acid sequence of the oligopeptide was identified as Gly-Pro-ALa-Gly-Hyp-Val-Gly-Ala-Hyp-Gly-Gly-Hyp-Gly-Val-Arg-Gly-Leu-Hyp-Gly-Asp-Arg-Gly. The results revealed that its amino acid sequence had strong homology to various types of collagen. Conclusion: Whitide might be an orally anticoagulant for its hot and trypsin stable.
  8 4,601 137
Optimization of ultrasonic-assisted extraction of antioxidant compounds from Guava (Psidium guajava L.) leaves using response surface methodology
Fansheng Kong, Shujuan Yu, Zeng Feng, Xinlan Wu
Jul-Sep 2015, 11(43):463-469
DOI:10.4103/0973-1296.160451  PMID:26246720
Objective: To optimization of extraction of antioxidant compounds from guava (Psidium guajava L.) leaves and showed that the guava leaves are the potential source of antioxidant compounds. Materials and Methods: The bioactive polysaccharide compounds of guava leaves (P. guajava L.) were obtained using ultrasonic assisted extraction. Extraction was carried out according to Box Behnken central composite design, and independent variables were temperature (20–60°C), time (20–40 min) and power (200–350 W). The extraction process was optimized by using response surface methodology for the highest crude extraction yield of bioactive polysaccharide compounds. Results: The optimal conditions were identified as 55°C, 30 min, and 240 W. 1,1-diphenyl-2-picryl-hydrazyl and hydroxyl free radical scavenging were conducted. Conclusion: The results of quantification showed that the guava leaves are the potential source of antioxidant compounds.
  6 3,951 191
Protective effect of calycosin-7-O-β-D-glucopyranoside against oxidative stress of BRL-3A cells induced by thioacetamide
Li Jian, Lin Xin, Ma Yufang, Huang Yifan
Jul-Sep 2015, 11(43):524-532
DOI:10.4103/0973-1296.160461  PMID:26246727
Background: Calycosin 7-O-β-D glucopyranoside (CG) is a natural isoflavone found in traditional Chinese medicines Astragali Radix (AR). Objective: Calycosin 7-O-β-D glucopyranoside, an isoflavone isolated from AR, has been found to have potent antioxidantive effects. This study was designed to investigate whether CG prevents oxidative stress induced by thioacetamide (TAA). Materials and Methods: BRL-3A cells were pretreated with different concentrations of CG (10, 20, 40 mg/mL) for 12 h and then exposed to 0.18 mol/L TAA for 2 h. The cell viability were examined by 3 [4,5 dimethylthiazol 2 yl] 2,5 diphenyl tetrazolium assay, total antioxidant capacity, malondialdehyde (MDA) and the activity of antioxidant enzymes, including catalase, glutathione peroxidase and superoxide dismutase were determined by microplate method. Reactive oxygen species (ROS) generation was quantified by the 2’,7’ dichlorofluorescin diacetate method. Protein and mRNA expression of CYP2E1 were determined by western blotting and real time PCR. Results: The cell oxidative stress was significantly increased after 2 h of TAA exposure. Pretreatment of BRL 3A cells with CG significantly increased the activities of antioxidant enzymes, scavenged ROS and reduced MDA production. CG decreased the expression of CYP2E1, and ultimately decreased TAA induced BRL 3A cells oxidative stress. Conclusions: Calycosin 7-O-β-D glucopyranoside has a protective effect against TAA induced oxidative stress in BRL 3A cells, and that the underlying mechanism involves in scavenging of ROS and the modulating expression of CYP2E1.
  6 3,151 115
Effect of modified Bo-yang-Hwan-o-Tang, a polyherbal medicine on the hippocampal neuronal damage in a rat model of global ischemia
Tae Woo Oh, Hyo Won Jung, Yong-Ki Park
Jul-Sep 2015, 11(43):665-673
DOI:10.4103/0973-1296.160445  PMID:26246747
Background: Chronic cerebral hypoperfusion has been well characterized as a common pathological status contributing to vascular dementia (VD). In this study, the neuroprotective effect of modified Bo yang Hwan O Tang (mBHT), a polyherbal medicine for ischemic stroke, was investigated in a rat model for global ischemia. Materials and Methods: Global ischemia model was prepared in Sprague Dawley rats by the permanent occlusion of bilateral common carotid arteries (two vessel occlusion [2VO]) induced chronic cerebral hypoperfusion. mBHT at doses of 250 and 500 mg/kg was orally administrated for 4 weeks once a day, 24 h after 2VO. Histopathological change of the hippocampal region was observed by hematoxylin and eosin, Nissl, and Fluoro Jade B staining and immunohistochemistry with anti glial fibrillary acidic protein and anti neuronal nuclei antibodies. The expression of Bax, Bcl 2, and caspase 3 was investigated in the hippocampus by Western blot. The nuclear factor kappa B (NF κB) expression was also analyzed in hippocampal CA1 region using immunofluorescence staining. Results: The administration of mBHT at doses of 250 and 500 mg/kg significantly inhibited chronic cerebral hypoperfusion induced neuronal damage and astroglial activation in the hippocampal CA1 region in 2VO rats. mBHT increased the NF κB expression in the CA1 neuronal cells but decreased in activated astrocytes. In addition, mBHT significantly decreased the hippocampal expression of Bax and caspase 3 and increased the Bcl 2 expression in 2VO rats. Conclusions: Our data indicate that mBHT has a neuroprotective property in VD induced by chronic cerebral hypoperfusion through inhibiting the hippocampal neuronal damage and astrogliosis.
  6 3,209 94
On-line quantitative monitoring of liquid-liquid extraction of Lonicera japonica and Artemisia annua using near-infrared spectroscopy and chemometrics
Sha Wu, Ye Jin, Qian Liu, Qi-an Liu, Jianxiong Wu, Yu-an Bi, Zhengzhong Wang, Wei Xiao
Jul-Sep 2015, 11(43):643-650
DOI:10.4103/0973-1296.160465  PMID:26246744
Background: Liquid liquid extraction of Lonicera japonica and Artemisia annua (JQ) plays a significant role in manufacturing Reduning injection. Many process parameters may influence liquid liquid extraction and cause fluctuations in product quality. Objective: To develop a near infrared (NIR) spectroscopy method for on line monitoring of liquid liquid extraction of JQ. Materials and Methods: Eleven batches of JQ extraction solution were obtained, ten for building quantitative models and one for assessing the predictive accuracy of established models. Neochlorogenic acid (NCA), chlorogenic acid (CA), cryptochlorogenic acid (CCA), isochlorogenic acid B (ICAB), isochlorogenic acid A (ICAA), isochlorogenic acid C (ICAC) and soluble solid content (SSC) were selected as quality control indicators, and measured by reference methods. NIR spectra were collected in transmittance mode. After selecting the spectral sub ranges, optimizing the spectral pretreatment and neglecting outliers, partial least squares regression models were built to predict the content of indicators. The model performance was evaluated by the coefficients of determination (R2), the root mean square errors of prediction (RMSEP) and the relative standard error of prediction (RSEP). Results: For NCA, CA, CCA, ICAB, ICAA, ICAC and SSC, R2 was 0.9674, 0.9704, 0.9641, 0.9514, 0.9436, 0.9640, 0.9809, RMSEP was 0.0280, 0.2913, 0.0710, 0.0590, 0.0815, 0.1506, 1.167, and RSEP was 2.32%, 4.14%, 3.86%, 5.65%, 7.29%, 6.95% and 4.18%, respectively. Conclusion: This study demonstrated that NIR spectroscopy could provide good predictive ability in monitoring of the content of quality control indicators in liquid liquid extraction of JQ.
  5 3,527 92
Application of microscopy technique and high performance liquid chromatography for quality assessment of the flower bud of Tussilago farfara L. (Kuandonghua)
Da Li, Li Liang, Jing Zhang, Tingguo Kang
Jul-Sep 2015, 11(43):594-600
DOI:10.4103/0973-1296.160473  PMID:26246737
Background: Quality control is one of the bottleneck problems limiting the application and development of traditional Chinese medicine (TCM). In recent years, microscopy and high performance liquid chromatography (HPLC) techniques have been frequently applied in the quality control of TCM. However, studies combining conventional microscopy and HPLC techniques for the quality control of the flower bud of Tussilago farfara L. (Kuandonghua) have not been reported. Objective: This study was undertaken to evaluate the quality of the flower bud of T. farfara L. and to establish the relationships between the quantity of pollen grains and four main bioactive constituents: tussilagone, chlorogenic acid, rutin and isoquercitrin. Materials and Methods: In this study, microscopic examination was used to quantify microscopic characteristics of the flower bud of T. farfara L., and the chemical components were determined by HPLC. The data were analyzed by Statistical Package for the Social Sciences statistics software. Results: The results of the analysis showed that tussilagone, chlorogenic acid, rutin and isoquercitrin were significantly correlated with the quantity of pollen grains in the flower bud of T. farfara L. There is a positive correlation between them. From these results, it can be deduced that the flower bud of T. farfara L. with a greater quantity of pollen grains should be of better quality. Conclusion: The study showed that the established method can be helpful for evaluating the quality of the flower bud of T. farfara L. based on microscopic characteristic constants and chemical quantitation.
  5 3,167 113
Simultaneous quantification of crocetin esters and picrocrocin changes in Chinese saffron by high-performance liquid chromatography-diode array detector during 15 years of storage
Yingpeng Tong, Yongqiu Yan, Xingyi Zhu, Ruoxi Liu, Feng Gong, Ling Zhang, Ping Wang
Jul-Sep 2015, 11(43):540-545
DOI:10.4103/0973-1296.160467  PMID:26246729
Background: Saffron, which is made up of the dried stigmas of Crocus sativus L., has been successfully cultivated in China since 1970s and Zhejiang province is now the largest producing area in China, but the contents of crocetin esters and picrocrocin in saffron from Zhejiang province has not been determined simultaneously by high performance liquid chromatography (HPLC) and changes of these constituents in Chinese saffron during storage for years has not been studied. Object: To establish a simple method quantification of the five main compounds including picrocrocin and four crocetin esters in saffron from main producing areas of China and study the influence of storage time on the changes of saffron constituents. Materials and Methods: A simple, sensitive, and accurate HPLC method was developed for simultaneous determination of five major active components in saffron and eight samples which collected from the same farm of Zhejiang province in different years were analyzed. Results: The correlation coefficient values (R2 > 0.9997) indicated good correlations between the investigated compounds’ concentrations and their peak areas within the test ranges. The limits of quantification and detection of the five compounds were 0.53–2.76 μg/mL and 0.11–0.77 μg/mL, respectively. The recoveries ranged from 94.67% to 101.31%, and the overall relative standard deviations for intra day and inter day were lower than 3.49%.The method was applied to study the changes of crocetin esters and picrocrocin contents in saffron samples during 15 years of storage. The losses of crocetin esters and picrocrocin in saffron with 1 year storage were 52.2% and 54.3%, respectively. The trend then declined during subsequent storage. Conclusion: The developed method can be applied to the intrinsic quality control of saffron.
  5 4,083 141
Microwave assisted extraction of polysaccharides from Yupingfeng powder and their antioxidant activity
Dan Wang, Bi-Bo Zhang, Xiao-Xia Qu, Feng Gao, Min-Yong Yuan
Jul-Sep 2015, 11(43):546-554
DOI:10.4103/0973-1296.160468  PMID:26246730
Background: Microwave assisted reflux extraction of polysaccharides YPF-P from the famous Chinese traditional drug, Yupingfeng powder, optimization of extracting conditions and evaluation of their antioxidant activity were conducted in this study. Results: Single factor effect trends were achieved through yields and contends of YPF P obtained from different extracting conditions. Then through a three level, four variable Box Behnken design of response surface methodology adopting yield as response, the optimal conditions were determined as follows: Material/solvent ratio 1:23.37, microwave power 560 W, Extraction temperature 64°C, and extraction time 9.62 min. Under the optimal conditions, the YPF P extraction yield was 3.23%, and its content was detected as 38.52%. In antioxidant assays, the YPF P was tested to possess 1,1 diphenyl 2 picrylhydrazyl (DPPH) radical scavenging activities with an IC50 value of 0.262 mg/ml. In addition, YPF P was also proved to have relatively low ferric reducing antioxidant power (FRAP), compared to Vc, through FRAP assay. Conclusion: In the microwave assisted reflux extraction research, good YPF P yield was achieved from materials with relatively low YPF P content. And for the first time, both DPPH and FRAP assays were conducted on YPF P, which proved that the antioxidant activity of YPF P contributed to the functions of this medicine.
  4 3,224 119
The ethanolic extract of Juglans sinensis leaves and twigs attenuates CCl4-induced hepatic oxidative stress in rats
Heejung Yang, Sang Hyun Sung, Young Choong Kim
Jul-Sep 2015, 11(43):533-539
DOI:10.4103/0973-1296.160463  PMID:26246728
Background: The nuts of Juglans sinensis Dode, walnut tree, are rich in unsaturated fatty acids and bioactive compounds with antioxidant activity on liver damages. However, hepatoprotective activity of the leaves and twigs of J. sinensis have not intensively studied yet. Objective: Hepatoprotective activity of the refined ethanolic extract of J. sinensis (JSE3) was evaluated using carbon tetrachloride (CCl4) intoxicated rats. Materials and Methods: Hepatotoxicity was induced in Sprague Dawley rats by intraperitoneal injection of CCl4 for 6 weeks in the presence or absence of JSE3 (100 and 200 mg/kg body weight). The hepatoprotective activity of JSE3 was assessed by biochemical parameters including plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT), and antioxidant enzymes, such as superoxide dismutase (SOD), glutathione reductase, glutathione peroxide, reduced glutathione and oxidized glutathione, along with histopathological studies on hepatic tissue. Results: JSE3 significantly decreased the elevated levels of AST and ALT and restored the reduced levels of antioxidant enzymes. JSE3 also decreased the amounts of collagen content accumulated by CCl4 intoxication. Conclusion: These results suggested that the refined extract of J. sinensis may have a potential to be developed as a therapeutic agent to treat hepatic diseases, such as fatty liver and hepatic fibrosis.
  4 2,784 81
Selective and cost effective protocol to separate bioactive triterpene acids from plant matrices using alkalinized ethanol: Application to leaves of Myrtaceae species
Adélia M. Belem Lima, Antonio Carlos Siani, Marcos Jun Nakamura, Luiz Antonio D’Avila
Jul-Sep 2015, 11(43):470-476
DOI:10.4103/0973-1296.160453  PMID:26246721
Background: Triterpenes as betulinic (BA), oleanolic (OA) and ursolic acids (UA) have increasingly gained therapeutic relevance due to their wide scope of pharmacological activities. To fit large scale demands, exploitable sources of these compounds have to be found and simple, cost effective methods to extract them developed. Leaf material represents the best plant sustainable raw material. To obtain triterpene acid rich extracts from leaves of Eugenia, Psidium and Syzygium species (Myrtaceae) by directly treating the dry plant material with alkalinized hydrated ethanol. This procedure was adapted from earlier methods to effect depolymerization of the leaf cutin. Materials and Methods: Extracts were prepared by shaking the milled dry leaves in freshly prepared 2% NaOH in 95% EtOH solution (1:4 w/v) at room temperature for 6 h. Working up the product in acidic aqueous medium led to clear precipitates in which BA, OA and UA were quantified by gas chromatography. Results: Pigment free and low polyphenol content extracts (1.2–2.8%) containing 6–50% of total triterpene acids were obtained for the six species assayed. UA (7–20%) predominated in most extracts, but BA preponderated in Eugenia florida (39%). Carried out in parallel, n hexane defatted leaves led to up to 9% enhancement of total acids in the extracts. The hydroalcoholate treatment of Myrtaceae species dry leaves proved to be a cost effective and environmentally friendly method to obtain triterpene acids, providing them be resistant to alkaline medium. These combined techniques might be applicable to other plant species and tissues.
  4 3,391 144
Three statistical experimental designs for enhancing yield of active compounds from herbal medicines and anti motion sickness bioactivity
Yan Chen, Cuiping Zhang, Mei Zhang, Xiaobing Fu
Jul-Sep 2015, 11(43):435-443
DOI:10.4103/0973-1296.160444  PMID:26246716
Background: Since antiquity, Zingiber officinale (ginger), pogostemonis herba, and radix aucklandiae have been used as traditional Chinese medicines to remit gastrointestinal discomfort. Recent evidences also show the efficacy of the three herbal medicines against nausea and vomiting. Objective: To optimize the CO2 supercritical fluid extraction (SFE CO2) conditions for ginger and the ethanol reflux extraction conditions for radix aucklandiae, control the quality of pogostemonis herba essential oil, and evaluate anti motion sickness activity of the compound recipes composed of the three herbal medicine extracts. Materials and Methods: Two orthogonal array designs L9 (3)4 were employed to optimize the SFE CO2 conditions for enhancing yield of 6 gingerol from ginger and the ethanol reflux extraction conditions for enhancing yield of costunolide and dehydrocostus lactone from radix aucklandiae; a uniform design U5(53) was applied for evaluation of anti motion sickness activity of the compound recipes. Results: Extraction pressure (P < 0.01), extraction temperature and extraction time (P < 0.05) have significant effects on the yield of 6 gingerol from ginger by SFE CO2; ethanol concentration (P < 0.01) and times of repeating extraction (P < 0.05) have significant effects on the total yield of costunolide and dehydrocostus lactone from radix aucklandiae by ethanol reflux extraction; the anti motion sickness effects of the optimized compound recipe composed of the three herbal medicine extracts were markedly better than those of dimenhydrinate. Conclusion: The compound recipe composed of ginger, pogostemonis herba, and radix aucklandiae could be developed as a promising anti motion sickness medicine.
  4 4,059 143
Total saponins from Discorea nipponica makino ameliorate urate excretion in hyperuricemic rats
Qi Zhou, Dong-Hua Yu, Shu-Min Liu, Ying Liu
Jul-Sep 2015, 11(43):567-573
DOI:10.4103/0973-1296.160442  PMID:26246733
Objective: The objective was to study the mechanism of reducing level of the uric acid by rhizoma dioscoreae nipponese. Materials and Methods: A total of 40 rats were divided into four groups: A normal group, hyperuricemia group, benzbromarone group (9 mg/kg) and total saponins from rhizoma dioscoreae nipponese (TDN) group (40 mg/kg). Adenine (100 mg/kg) and ethambutol (250 mg/kg) were used to induce hyperuricemic rats. Immunohistochemical and Western blotting methods were used to detect the mRNA and proteins expressions of rat organic anion transporter1 (rOAT1), rat organic anion transporter3 (rOAT3) and rat urate transporter1 (rURAT1) in the kidneys of different groups. Results: It was found that the reduced concentration of blood uric acid was due to the enhancement of renal uric acid excretion. It was realized by up regulating proteins expressions of rOAT1 and rOAT3 and down regulating of rURAT1. Conclusion: The findings suggested that there were uricosuric effects of TDN by regulating renal organic ion transporters in hyperuricemic animals. Altogether, TDN may be a good Chinese herb in treating hyperuricemia, even a potential drug for gouty arthritis.
  4 3,135 126
Neuroprotective activity of parawixin 10, a compound isolated from Parawixia bistriata spider venom (Araneidae: Araneae) in rats undergoing intrahippocampal NMDA microinjection
Helene Aparecida Fachim, Marcia Renata Mortari, Leonardo Gobbo-Netto, Wagner Ferreira dos Santos
Jul-Sep 2015, 11(43):579-585
DOI:10.4103/0973-1296.160450  PMID:26246735
Background: Parawixia bistriata is a semi colonial spider found mainly in southeastern of Brazil. Parawixin 10 (Pwx 10) a compound isolated from this spider venom has been demonstrated to act as neuroprotective in models of injury regulating the glutamatergic neurotransmission through glutamate transporters. Objectives: The aim of this work was to evaluate the neuroprotective effect of Pwx 10 in a rat model of excitotoxic brain injury by N methyl D aspartate (NMDA) injection. Material and Methods: Male Wistar rats have been used, submitted to stereotaxic surgery for saline or NMDA microinjection into dorsal hippocampus. Two groups of animals were treated with Pwx 10. These treated groups received a daily injection of the Pwx 10 (2.5 mg/μL) in the right lateral ventricle into rats pretreated with NMDA, always at the same time, each one starting the treatment 1 h or 24 h. Nissl staining was performed for evaluating the extension and efficacy of the NMDA injury and the neuroprotective effect of Pwx 10. Results: The treatment with Pwx 10 showed neuroprotective effect, being most pronounced when the compound was administrated from 1 h after NMDA in all hippocampal subfields analyzed (CA1, CA3 and hilus). Conclusion: These results indicated that Pwx 10 may be a good template to develop therapeutic drugs for treating neurodegenerative diseases, reinforcing the importance of continuing studies on its effects in the central nervous system.
  4 3,109 92
A high performance liquid chromatography with ultraviolet method for Eschweilera nana leaves and their anti-inflammatory and antioxidant activities
Priscila M. Outuki, Nides S. Lazzeri, Lizziane M. B. de Francisco, Ciomar A. Bersani Amado, Izabel C. P. Ferreira, Mara Lane C. Cardoso
Jul-Sep 2015, 11(43):619-626
DOI:10.4103/0973-1296.160464  PMID:26246741
Background: Eschweilera nana Miers is a tree widely distributed in Cerrado, Brazil. Objective: In this study, we aimed to describe its phytochemical properties and antioxidant and topical anti inflammatory effects for the first time, as well validate an high performance liquid chromatography with ultraviolet/visible (HPLC UV Vis) method for the separation and quantification of the main components (hyperoside and rutin) in the hydroalcoholic extract of E. nana leaves. Materials and Methods: Structural identification of compounds in E. nana extract was performed by analysis of spectral data by 1H nuclear magnetic resonance, 13C nuclear magnetic resonance and/or ESI/EM. The HPLC UV Vis method was validated according International Conference on Harmonization (ICH) parameters. The 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) method were used for determination of in vitro antioxidant activities and the croton oil induced inflammation for evaluation of in vivo anti inflammatory effects. Results: Hyperoside, rutin, α-amirin, β-amirin, β-sitosterol, and stigmasterol were identified in the hydroalcoholic extract of E. nana leaves. HPLC UV Vis was validated according to ICH parameters. Furthermore, in vitro and in vivo assays demonstrated that the hydroalcoholic extract and methanol fraction showed significant antioxidant and topical anti inflammatory effects, as they were able to reduce ear edema induced by croton oil application. Conclusions: This research showed the first phytochemical study of E. nana extract and their biological activities may be associated with the presence of flavonoids in the extracts.
  3 3,792 184
Quantification of maltol in Korean ginseng ( Panax ginseng) products by high-performance liquid chromatography-diode array detector
Hyun Cheol Jeong, Hee-Do Hong, Young-Chan Kim, Young Kyoung Rhee, Sang Yoon Choi, Kyung-Tack Kim, Sung Soo Kim, Young-Chul Lee, Chang-Won Cho
Jul-Sep 2015, 11(43):657-664
DOI:10.4103/0973-1296.160452  PMID:26246746
Background: Maltol, as a type of phenolic compounds, is produced by the browning reaction during the high temperature treatment of ginseng. Thus, maltol can be used as a marker for the quality control of various ginseng products manufactured by high temperature treatment including red ginseng. For the quantification of maltol in Korean ginseng products, an effective high performance liquid chromatography diode array detector (HPLC DAD) method was developed. Materials and Methods: The HPLC DAD method for maltol quantification coupled with a liquid liquid extraction (LLE) method was developed and validated in terms of linearity, precision, and accuracy. An HPLC separation was performed on a C18 column. Results: The LLE methods and HPLC running conditions for maltol quantification were optimized. The calibration curve of the maltol exhibited good linearity (R2 = 1.00). The limit of detection value of maltol was 0.26 μg/mL, and the limit of quantification value was 0.79 μg/mL. The relative standard deviations (RSDs) of the data of the intra and inter day experiments were <1.27% and 0.61%, respectively. The results of the recovery test were 101.35–101.75% with an RSD value of 0.21–1.65%. The developed method was applied successfully to quantify the maltol in three ginseng products manufactured by different methods. Conclusion: The results of validation demonstrated that the proposed HPLC DAD method was useful for the quantification of maltol in various ginseng products.
  3 3,172 88
A study of pharmacokinetic interactions among co-existing ingredients in Viscum coloratum after intravenous administration of three different preparations to rats
Yuying Ma, Ronghua Fan, Mengmeng Duan, Zhiguo Yu, Yunli Zhao
Jul-Sep 2015, 11(43):455-462
DOI:10.4103/0973-1296.160448  PMID:26246719
Background: Viscum coloratum (Komar) Nakai, known as Hujisheng in china, has been widely used as a herb medicine to treat a variety of diseases, including cardiovascular diseases, cancer, hypertension, hepatitis and hemorrhage. Objective: The aim was to investigate pharmacokinetic interactions among co existing ingredients in V. coloratum after intravenous administration of three different preparations (four monomer solutions, the mixture of them and Viscum coloratum extracts) to rats. Materials and Methods: After protein precipitation pretreatment with plasma samples, high performance liquid chromatographic methods were developed and applied to quantitatively determinate the four components [syringin (Syri), homoeriodictyol 7 O-β-D glycoside (Hedt III), homoeriodictyol 7 O-β-D apiose (1 → 2)-β-D glycoside (Hedt II) and homoeriodictyol 7 O-β-D apiosiyl (1 → 5)-β-D apiosyl (1 → 2)-β-D glycoside (Hedt I)].The pharmacokinetic parameters (Area under the curve [AUC (0-t)], AUC (0-∞), t1/2) were calculated using DAS 2.1 software (Chinese Pharmacological Society,Shanghai,China) and compared statistically by One way analysis of variance using SPSS software (18.0, Chicago, IL, USA) with P < 0.05 considered statistically significant. Results: Good linearities were achieved in the measured concentration range with R2 it0.9920. Precision, accuracy and extraction recovery were all within the acceptable range. For Syri, there was a significant difference only on t1/2 among three treatment groups. For Hedt I, Hedt II and Hedt III, three flavonoid glycosides, the change of AUC (0-t), AUC (0-∞) and t1/2 were markedly distinctive and even converse. Conclusion: Complex, extensive pharmacokinetic interactions were observed among these components in V. coloratum. They were mutually influenced by the in vivo absorption, distribution, metabolism and elimination. The result suggested traditional Chinese medicine was a complicated system, and we should take a scientific and dialectic view in the research and development processes.
  3 3,266 100
Simultaneous determination of nine marker compounds in the traditional Korean medicine, Dangguisu san by high performance liquid chromatography
Chang-Seob Seo, Hyeun-Kyoo Shin
Jul-Sep 2015, 11(43):555-561
DOI:10.4103/0973-1296.160457  PMID:26246731
Background: Dangguisu-san (DGSS) has been widely used to treat ecchymosis, blood stagnation and pain resulting from physical shock in Korea. Objective: A high performance liquid chromatography–photodiode array detection (HPLC–PDA) method for simultaneous analysis of nine components, albiflorin (1), paeoniflorin (2), liquiritin (3), nodakenin (4), coumarin (5), liquiritigenin (6), cinnamic acid (7), cinnamaldehyde (8), and glycyrrhizin (9) in DGSS extract has been developed for the first time. Materials and Methods: The analytical column for separation of the nine constituents used a Gemini C18 column kept at 40°C by the gradient elution with 1.0% (v/v) acetic acid in water and 1.0% (v/v) acetic acid in acetonitrile as mobile phase. The flow rate was 1.0 mL/min and the injection volume was 10 μL. Results: Calibration curves of all compounds showed good linearity (r2 ≥ 0.9999) within the test ranges. The limits of detection and quantification for all analytes were 0.01–0.27 μg/mL and 0.04–0.89 μg/mL, respectively. All recoveries of the nine marker compounds were 96.62–102.47% with relative standard deviations (RSD) <1.72%. The RSDs of intra day and inter day precision were <1.32% and 1.61%, respectively. The amounts of the nine marker components ranged from 0.10 mg/g to 13.71 mg/g. Conclusion: The developed and validated HPLC–PDA method may help for the quality control of DGSS.
  2 3,060 90
Simultaneous determination of three sesquiterpene lactones in Aucklandia lappa Decne by high performance liquid chromatography
Chang-Seob Seo, Hyeun-Kyoo Shin
Jul-Sep 2015, 11(43):562-566
DOI:10.4103/0973-1296.160471  PMID:26246732
Background: Aucklandia lappa Decne, a well known traditional herbal medicine, is used for the treatment of asthma, rheumatism, coughs, tuberculosis, and many other diseases. Objective: We performed simultaneous analysis of three sesquiterpene lactones, costunolide (1), dehydrocostus lactone (2), and alantolactone (3), obtained from a 70% methanol extract of A. lappa using high performance liquid chromatography–photodiode array (HPLC–PDA) techniques. Materials and Methods: The compounds 1–3 were separated using a reversed phase SunFire C18 analytical column kept at 35°C by the isocratic elution with distilled water and acetonitrile as mobile phase. The flow rate was 1.0 mL/min, and the injection volume was 10 μL. Results: The established analytical method showed high linearity, with a correlation coefficient ≥0.9999. The limit of detection and the limit of quantification of compounds 1–3 were 0.06–0.13 μg/mL and 0.21–0.42 μg/mL, respectively. The recovery of the compounds 1–3 was 97.27–103.00%. The intra and inter day relative standard deviations were 0.09–0.97% and 0.09–1.06%, respectively. The amounts of the compounds 1–3 were 17.32, 28.26, and 0.01 mg/g, respectively. Conclusion: The established and validated HPLC–PDA method may be help for the quality control of herbal medicine, A. lappa.
  2 3,085 128
Optimization of induction, subculture conditions, and growth kinetics of Angelica sinensis (Oliv.) Diels callus
Bing Huang, Lijuan Han, Shaomei Li, Chunyan Yan
Jul-Sep 2015, 11(43):574-578
DOI:10.4103/0973-1296.160443  PMID:26246734
Background: Angelica sinensis (Oliv.) Diels is an important traditional Chinese medicine, and the medicinal position is its root. This perennial herb grows vigorously only in specific areas and the environment. Tissue culture induction of callus and plant regeneration is an important and effective way to obtain large scale cultures of A. sinensis. Objective: The objective was to optimize the inductive, subculture conditions, and growth kinetics of A. sinensis. Materials and Methods: Tissue culture conditions for A. sinensis were optimized using leaves and petioles (types I and II) as explants source. Murashige and Skoog (MS) and H media supplemented with 30 g/L sucrose, 7.5 g/L agar, and varying concentrations of plant growth regulators were used for callus induction. In addition, four different basal media supplemented with 1.0 mg/L 2,4 dichlorophenoxy acetic acid (2,4 D), 0.2 mg/L 6 benzyladenine (BA) and 30 g/L sucrose were optimized for callus subculture. Finally, growth kinetics of A. sinensis cultured on different subculture media was investigated based on callus properties, including fresh weight, dry weight, medium pH, callus relative fresh weight growth, callus relative growth rate (CRGR), and sucrose content. Results: MS medium supplemented with 5 mg/L α-naphthaleneacetic acid, 0.5 mg/L BA, 0.7 mg/L 2,4 D, 30 g/L sucrose and 7.5 g/L agar resulted in optimal callus induction in A. sinensis while petiole I was found as the best plant organ for callus induction. The B5 medium supplemented with 1.0 mg/L 2,4 D, 0.2 mg/L BA and 30 g/L sucrose displayed the best results in A. sinensis callus subculture assays. Conclusion: The optimized conditions could be one of the most potent methods for large scale tissue culture of A. sinensis.
  2 3,867 410
Region-selective biosynthesis of artemisinic acid glycosides by crown galls of Panax quinquefolium and their in vitro antitumor activities
Jianhua Zhu, Lijia Chen, Xianjing Hu, Liyan Song, Mingxuan Wang, Rongmin Yu
Jul-Sep 2015, 11(43):518-523
DOI:10.4103/0973-1296.160460  PMID:26246726
Background: The biosynthesis of artemisinin derivatives is one of the interesting subjects. Artemisinic acid (AA) has been widely studied as a supposed intermediate in the biosynthetic pathway leading to artemisinin in Artemisia annua. Objective: To investigate the bioconversion of AA by transgenic crown galls of Panax quinquefolium. Materials and Methods: AA was administered into crown galls of P. quinquefolium and co cultured for 2 days. The methanol extract was separated by column chromatography, and the structures of two biosynthesis products were elucidated by physicochemical and spectroscopic methods. Co culture time curves on conversion were also established. In addition, the effects of AA on the growth and ginsenosides production of crown galls of P. quinquefolium were investigated. Furthermore, the in vitro antitumor activities of AA and two glycosides against HepG2 cell line were evaluated by 3 (4,5 dimethylthiazol 2 yl) 2,5 diphenyl tetrazolium bromide (MTT) assay. Results: Glycosylation of AA by crown galls of P. quinquefolium was observed, and two region selectively glycosylated products were obtained (AA-1, AA-2), involving one new compound (AA-2). Their structures were elucidated to be AA β-D-glucopyranosyl ester (AA-1) and AA β-D-glucopyranosyl (2 → 1) β-D-glucopyranosyl ester (AA 2). The maximum yield of AA 1 was 19.3% on the 1st co culture day while that of AA 2 was 59.1% on the 2nd day. MTT assay showed that the activity of monosaccharide glycoside (AA 1) was better than that of disaccharide glycoside (AA 2). Conclusion: Two AA glycosides involved one new compound with potential antitumor activity were obtained by region selective biosynthesis with crown galls of P. quinquefolium.
  1 2,620 84
Biological activities of Croton palanostigma Klotzsch
Eduardo Ferreira Mota, Diele Magno Rosario, Andreza Socorro Silva Veiga, Davi Do Socorro Barros Brasil, Fernando Tobias Silveira, Maria Fâni Dolabela
Jul-Sep 2015, 11(43):601-606
DOI:10.4103/0973-1296.160449  PMID:26246738
Background: Different species of Croton are used in traditional Amazonian medicine. Among the popular uses are treatment of bacterial diseases, poorly healing wounds and fevers. Objective: This study evaluated the antileishmanial, antiplasmodial and antimicrobial activities of the extracts and diterpenes of Croton palanostigma Klotzsch (Euphorbiaceae). Materials and Methods: Leaves and bark were extracted with dichloromethane and methanol. The bark dichloromethane extract (BDE) was chromatographed on a column, obtaining cordatin and aparisthman. The extracts and diterpenes were assayed thought agar disk diffusion method and their bactericidal or fungicidal effects were evaluated by minimum bactericidal or fungicidal concentration. The antiplasmodial activity was evaluated after 24 and 72 h of exposition. The antileishmanial activity was performed on promastigotes forms of Leishmania amazonensis. Results: The bark methanol extract (BME) and cordatin were not active against any microbial strains tested; BDE and leaves methanol extract (LME) were positive for Pseudomonas aeruginosa and aparisthman was positive for Candida albicans. In the determination of the minimum bactericidal concentration, neither of them were active in the highest concentration tested. The extracts and diterpenes were inactive in Plasmodium falciparum, except the LME in 72 h. Any extract was shown to be active in promastigote forms of L. amazonensis. Conclusion: These results indicate that the BDE and LME did not inhibit the bacterial growth, then they probably had bacteriostatic effect. LME presented activity in P. falciparum.
  1 2,742 87
Traditional medicine yanggyuksanhwa-tang inhibits adipogenesis and suppresses proliferator-activated receptor gamma expression in 3T3-L1 cells
Soo-Jin Jeong, Sae-Rom Yoo, Chang-Seob Seo, Hyeun-Kyoo Shin
Jul-Sep 2015, 11(43):502-508
DOI:10.4103/0973-1296.160456  PMID:26246724
Background: Yanggyuksanhwa-tang (YGSHT) is a specific traditional Korean herbal formula for Soyangin according to Sasang constitutional philosophy. Although its biological activities against inflammation and cerebral infarction have been reporting, there is no information about the adipogenic activity of YGSHT. In the present study, we investigated the anti adipogenic activity of YGSHT to evaluate effects of YGSHT on adipogenesis in vitro. Materials and Methods: Using 3T3 L1 preadipocytes, we induced the cellular differentiation into adipocytes by adding insulin. Anti adipogenic activity of YGSHT was measured by oil red O staining, triglyceride assay, glycerol 3 phosphate dehydrogenase (GPDH) activity test, and leptin assay. Results: YGSHT extract had no significant cytotoxicity in preadipocytes or differentiated adipocytes. YGSHT reduced the number of lipid droplets and content of triglyceride in adipose cells. YGSHT also significantly inhibited GPDH activity and decreased leptin production compared with control adipocytes. Down regulation of peroxisome proliferator activated receptor gamma (PPAR g) expression at the messenger RNA level was observed in YGSHT treated adipocytes. Conclusion: Taken together, our data suggest that YGSHT has potential as an anti-obesity drug candidate.
  - 2,933 84
Simultaneous determination of four active components in rat plasma by ultra-high performance liquid chromatography tandem-mass spectrometry/mass spectrometry and its application to a pharmacokinetic study after oral administration of Callicarpa nudiflora extract
Jun Shao, Shuangcheng Ma, Dongkun Zheng, Weikang Chen, Yuehua Luo
Jul-Sep 2015, 11(43):509-517
DOI:10.4103/0973-1296.160458  PMID:26246725
Background: Callicarpa nudiflora has been commonly used as a Chinese folk medicine for resolving toxin, dispersing edema and hemostasis; however, its pharmacokinetic (PK) behavior remains unknown. In our present study, a simple and sensitive ultra high performance liquid chromatography tandem mass spectrometry method was firstly developed on simultaneous determination and PK study of four active components (luteoloside, dracocephaloside, juncein and nudifloside) following the oral administration of C. nudiflora extract to investigate their PK profiles. Materials and Methods: Chromatographic separation was achieved on a Phenomenex® Kinetex C18 column (50 mm × 2.1 mm, 1.7 μm) with gradient elution using a mobile phase consisted of acetonitrile (A) and 0.05‰ formic acid in water (B). The quantitation was carried out by multiple reaction monitoring using electrospray ionization in the negative ion mode. Results: Calibration curves offered satisfactory linearity, with correlation coefficients >0.99 for all compounds within the concentration range. The low limits of quantification were 1.03 ng/mL for luteoloside, 1.16 ng/mL for dracocephaloside, 0.82 ng/mL for juncein and 0.88 ng/mL for nudifloside, respectively. The intra and inter day precisions (relative standard deviation) were within 7.4% and the accuracies (relative error) ranged from −7.4% to 7.9%. Conclusion: This method was successfully applied to the PK studies of luteoloside, dracocephaloside, juncein and nudifloside in rat plasma after oral administration of C. nudiflora extract, four analytes exhibited quick absorption with peak concentrations occurring at around 25 min and eliminated rapidly.
  - 2,940 86