Identification and evaluation of anti hepatitis C virus phytochemicals from Eclipta alba.
Identification and evaluation of anti hepatitis C virus phytochemicals from Eclipta alba.
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DOI：
10.1016/j.jep.2012.09.036
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发表时间：
2012-12-18
影响因子：
5.4
通讯作者：
Pandey, Virendra N.
中科院分区：
文献类型：
--
作者：
Manvar, Dinesh;Mishra, Mahesh;Kumar, Suriender;Pandey, Virendra N.
关键词：
在印度，牛乳草，传统上被称为牛乳草，已经在阿育吠陀医学中使用了1000多年。在印度，它被用于治疗感染性肝炎、肝硬化、肝肿大和其他肝脏和胆囊疾病。为了评估黄花提取物中存在的抗丙型肝炎病毒活性，进行基于生物测定的分离，并从活性部分中鉴定抗丙型肝炎病毒的植物化学物质。采用生物活性引导分馏法对活性成分进行鉴定。采用硅胶色谱和反相高效液相色谱相结合的方法分离活性分离物。研究了黄花提取物及其分离物在体外抑制HCV复制酶（HCV NS5B）活性和在携带复制HCV亚基因组RNA复制子的细胞培养系统中抑制HCV复制的能力。用荧光猝灭法检测纯化的分离株与丙型肝炎病毒复制酶的结合亲和力，用MTT法检测其细胞毒性。黄花提取物对体外HCV复制酶RdRp活性有较强的抑制作用。在细胞培养系统中，有效抑制HCV复制，降低HCV RNA滴度和病毒蛋白翻译水平。基于生物测定的提取物和活性部分中抗hcv植物化学物质的纯化鉴定出三种化合物：维地内酯、木犀草素和芹菜素。这些化合物在体外对HCV复制酶的抑制作用表现出剂量依赖性，并且在细胞培养体系中具有抗HCV复制活性，在体外和细胞培养体系中，从黄花提取物和活性部分分离的植物化学物质均表现出抗HCV复制活性。标准的白月牙提取物或其分离物可作为抗HCV的有效替代和补充治疗。
Eclipta alba, traditionally known as bhringraj, has been used in Ayurvedic medicine for more than 1000 years in India. It is used for the treatment of infective hepatitis, liver cirrhosis, liver enlargement and other ailments of liver and gall bladder in India. To evaluate anti-hepatitis C virus activity present in the Eclipta alba extract, perform bioassay based fractionation and identify anti-HCV phytochemicals from the active fractions. Identification of active compounds was performed by bio-activity guided fractionation approach. Active isolates were separated by the combination of silica gel chromatography and preparative scale reverse phase HPLC. Eclipta alba extract and its isolates were examined for their ability to inhibit HCV replicase (HCV NS5B) activity in vitro and HCV replication in a cell culture system carrying replicating HCV subgenomic RNA replicon. The purified isolates were also examined for their binding affinity to HCV replicase by fluorescence quenching and their cytotoxicity by MTT assay. Eclipta alba extract strongly inhibited RNA dependent RNA polymerase (RdRp) activity of HCV replicase in vitro. In cell culture system, it effectively inhibited HCV replication which resulted in reduced HCV RNA titer and translation level of viral proteins. Bioassay-based fractionations of the extracts and purification of anti-HCV phytochemicals present in the active fractions have identified three compounds, wedelolactone, luteolin, and apigenin. These compounds exhibited dose dependent inhibition of HCV replicase in vitro, and anti-HCV replication activity in the cell culture system Eclipta alba extract and phytochemicals isolated from active fractions display anti-HCV activity in vitro and in cell culture system. The standardized Eclipta alba extract or its isolates can be used as an effective alternative and complementary treatment against HCV.
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