基于网络药理学探讨丹参抗乙肝病毒的活性成分与作用机制

Investigation on anti-hepatitis B virus active components and mechanism of Salvia miltiorrhiza based on network pharmacology

应用网络药理学方法预测了丹参抗乙肝病毒(hepatitis B virus,HBV)的活性成分和作用机制。首先通过TCMSP、PubChem数据库及文献调研获取丹参的化学成分,通过SwissTargetPrediction和GeneCards数据库分别预测活性成分和HBV感染所对应的潜在作用靶点。使用String数据库构建蛋白互作关系网络、Cytoscape软件将成分-作用靶点网络可视化并进行拓扑学分析、DAVID平台进行GO功能及KEGG通路富集分析、AutoDock Vina软件将丹参关键活性成分与核心靶蛋白进行分子对接。共筛选出38种活性成分,获得疾病-化合物交集靶点178个,富集分析得到405个GO相关条目、68条相关信号通路(涉及T细胞/B细胞受体信号通路、PI3K/AKT信号通路及mTOR信号通路等)。分子对接结果显示,丹参大多数关键活性成分(丹参醌酚Ⅱ、丹参新酮、原儿茶酸、紫草酸和原儿茶醛)与核心靶点(PIK3CA、APP、STAT3、AKT1和mTOR)具有较好的亲和力。更进一步,在HBV复制小鼠模型上考察了丹参代表性活性成分紫草酸的抗HBV活性及对PI3K/AKT和mTOR信号通路的调控作用。动物福利和实验操作均遵循湖北大学动物伦理与福利委员会的规定。结果显示,紫草酸能明显抑制模型小鼠的HBV DNA复制,降低血清HBsAg和HBeAg水平,抑制肝脏AKT和mTOR的磷酸化,表明紫草酸可能通过调控PI3K/AKT及mTOR信号通路发挥抗HBV作用。

We predicted the anti-hepatitis B virus(HBV)active components and mechanism of Salvia miltiorrhiza based on network pharmacology.The active components of S.miltiorrhiza were obtained through TCMSP,PubChem database and literature research.The potential targets of the active components and HBV infection were predicted by SwissTargetPrediction and GeneCards databases,respectively.The protein-protein interaction(PPI)network was constructed by String database.Cytoscape software was adopted to construct a visual network of active component-disease target and perform topological analysis.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses were performed using DAVID platform.The molecular docking of key components and core targets was carried out by AutoDock Vina software.We screened out a total of 38 active components and 178 disease-component overlapping targets.Enrichment analyses obtained 405 related GO items and 68 signaling pathways,such as T/B cell receptor signaling pathways,PI3K/AKT signaling pathway,and mTOR signaling pathway.According to the results of molecular docking,most characteristic components of S.miltiorrhiza(miltiononeⅡ,miltirone,protocatechuic acid,lithospermic acid,protocatechualdehyde)showed good affinity with the key targets(PIK3CA,APP,STAT3,AKT1 and mTOR).Furthermore,the anti-HBV activity of lithospermic acid,the representative active component of S.miltiorrhiza,and its regulation on PI3K/AKT and mTOR signaling pathways were investigated in an HBV replicating mouse model.Animal welfare and experimental procedures follow the regulations of the Animal Ethics and Welfare Committee of Hubei University.The results showed that lithospermic acid significantly inhibited HBV DNA replication,reduced serum HBsAg and HBeAg levels,and decreased the phosphorylation protein expression levels of AKT and mTOR in liver,indicating that lithospermic acid might exert the anti-HBV activity by regulating PI3K/AKT and mTOR signaling pathways.