基于网络药理学和实验验证探讨苦参治疗乙型病毒性肝炎的作用机制

Mechanism study of Kushen(Sophorae Flavescentis Radix)in treating viral hepatitis type B based on network pharmacology and experimental test

目的基于网络药理学和分子对接技术,探讨苦参治疗乙型病毒性肝炎的作用机制,为临床防治提供指导。方法通过GeneCards、OMIM和TTD数据库获取乙型病毒性肝炎靶点,使用TCMSP、BATMAN-TCM、ETCM、HERB数据库及查阅相关文献获取苦参活性成分及靶点,并利用VENNY作图取两者的交集。用Cytoscape、STRING构建"活性成分-潜在靶点"网络图和蛋白互作网络图;应用DAVID数据库进行GO和KEGG通路的富集分析;最后通过分子对接和qPCR对上述结果进行验证。结果乙型病毒性肝炎靶标1827个,苦参活性成分39个,主要成分为刺芒柄花素、8-isopentenyl-kaempferol、(2R)-7-hydroxy-2-(4-hydroxyphenyl)chroman-4-one、高丽槐素、氧化苦参碱;苦参干预乙型病毒性肝炎共同靶点64个,其核心靶点为IL-6、AKT1、VEGFA、PTGS2、JUN、CASP3、TNF和MAPK1;KEGG富集得到通路111条,包括癌症的通路、乙型肝炎和TNF信号通路;分子对接技术显示氧化苦参碱与TNF、IL-6结合,结合能为-6.1~-9.8 kcal·mol;体外实验证实氧化苦参碱可下调HepG2.2.15细胞中TNF-α、IL-6 mRNA的表达,且随着剂量的增加,下调趋势更显著。结论苦参具有多成分、多靶点特性,其主要成分氧化苦参碱可通过激活TNF信号通路,下调TNF-α、IL-6 mRNA的表达,控制肝脏炎症,抑制HBV复制。

Objective To explore the potential mechanisms of action of Kushen(Sophorae Flavescentis Radix)against viral hepatitis type B,and to provide guidance for improvement in clinical prevention and treatment based on network pharmacology and molecular docking technology.Methods GeneCards,OMIM,and TTD databases were searched to obtain viral hepatitis type B targets.The active ingredients and targets of Kushen(Sophorae Flavescentis Radix)were searched through TCMSP,BATMAN-TCM,ETCM,HERB databases and related literature.The VENNY was used to map the intersection of the two search results.Cytoscape and STRING were used to construct the"active ingredient-potential target"network diagram and protein-protein interaction network diagram;then the DAVID database was used for the enrichment analysis of GO and KEGG pathways.Finally,molecular docking and qPCR were used to verify the above results.Results There were 1827 targets of viral hepatitis type B and 39 active components of Kushen(Sophorae Flavescentis Radix),including formononetin,8-isopentenyl-kaempferol,(2 R)-7-hydroxy-2-(4-hydroxyphenyl)chroman-4-one,sophorin,oxymatrine.There were 64 common targets for viral hepatitis type B and Kushen(Sophorae Flavescentis Radix),including core targets IL-6,AKT1,VEGFA,PTGS2,JUN,CASP3,TNF and MAPK1.KEGG analysis enriched 111 pathways,including cancer pathway,hepatitis type B and TNF signaling pathway.The molecular docking revealed that oxymatrine could bind with TNF and IL-6,with binding energy between-6.1~-9.8 kcal·mol;.Vitro experiments confirmed that oxymatrine can down-regulate the expression of TNF-αand IL-6 m RNA in HepG2.2.15 cells,and with the increase of the dose,the down-regulation trend was more significant.Conclusion Kushen(Sophorae Flavescentis Radix)has multicomponent and multi-target characteristics.Its main component oxymatrine can activate TNF signaling pathways,down-regulate the expression of TNF-αand IL-6 mRNA,control liver inflammation,and inhibit viral hepatitis type B replication.