基于网络药理学的“柴胡-黄芩”药对干预足细胞病变作用机制探索

Exploration of the Mechanism about Couplet Medicine of Bupleurum falcatum-Scutellaria baicalensis Intervening in Podocyte Lesion Based on Network Pharmacology

目的:预测辛通畅络法载体——复方肾苏Ⅱ之君药“柴胡-黄芩”药对干预足细胞病变的可能作用靶点与机制,为辛通畅络法防治足细胞病变序贯临床和基础研究的开展提供参考。方法:基于中药系统药理学技术平台(TCMSP)数据库,检索柴胡、黄芩化学成分及其对应靶点蛋白,借助Cytoscape 3.2.1软件绘制“中药-成分-靶点”网络图。在OMIM数据库、DrugBank数据库及DigSee在线文本中检索足细胞病变相关靶点,利用Venny 2.1.0在线作图工具获取其与“柴胡-黄芩”作用靶点的交集基因。分别应用STRING数据库及Cytoscape 3.2.1软件的“CytoNCA”插件构建交集基因蛋白质-蛋白质相互作用关系(PPI)网络图并进行拓扑学分析,获取核心预测靶点。借助DAVID数据库对交集基因进行基因本体(GO)功能注释及京都基因与基因组百科全书(KEGG)通路富集,并通过OmicShare Tools在线作图平台实现富集结果的可视化。结果:基于TCMSP数据库检索结果,获得有对应靶点的活性成分44种,其中柴胡13种、黄芩32种,豆甾醇为两者共有成分;潜在作用靶点较多的化合物为槲皮素、山柰酚、汉黄芩素等;节点度值较高靶点蛋白为前列腺素内过氧化物合酶2(PTGS2)、PTGS1、核受体共激活因子2、热休克蛋白90α等,分别与37、30、25、25个活性成分相关联。获取“柴胡-黄芩”作用靶点与足细胞病变相关靶点交集基因20个,包括PTGS2、VEGFA、MMP9、TNF、IL6等。上述交集基因的PPI网络图共包含节点20个、连线110条,MMP9、VEGFA、IL6等基因处于核心位置。GO分析结果显示,共获得生物信息条目154个(P<0.05),包括生物过程条目139个、细胞组成条目8个、分子功能条目7个。其中,生物过程主要涉及一氧化氮生物合成过程的正调控、炎症反应、免疫反应等,细胞组成主要涉及细胞外间隙、胞外区、质膜外侧等,分子功能主要涉及蛋白质结合、细胞因子活性、生长因子活性等。同时,获得KEGG通路富集条目47个(P<0.05),主要涉及细胞因子-细胞因子受体相互作用、类风湿关节炎、疟疾、癌症信号通路等相关信号转导途径。结论:复方肾苏Ⅱ之“柴胡-黄芩”药对活性成分可能通过细胞因子-细胞因子受体相互作用、类风湿关节炎、疟疾、癌症信号通路等相关信号转导途径作用于MMP9、VEGFA、IL6、TNF等靶点,从而发挥对足细胞病变的干预作用。

OBJECTIVE:To predict the potential target and mechanism of Xintong Changluo Method carrier-Compund ShensuⅡcouplet medicine of Bupleurum falcatum-Scutellaria baicalensis intervening in podocyte lesion,and to provide reference for the development of sequential clinical and basic research of Xintong Changluo Method in the prevention and treatment of podocyte lesion.METHODS:Based on TCMSP database,chemical components and target protein of B.falcatum and S.baicalensis were retrieved,and Cytoscape 3.2.1 software was used to draw a“TCM-component-target”network.The targets related to podocyte lesion were searched from OMIM database,DrugBank database and Digsee online text,and the intersection genes of above targets and“B.falcatum-S.baicalensis”target were obtained by Venny 2.1.0 online mapping tool.The protein-protein interaction(PPI)network was constructed by STRING database,and the core targets were obtained by topology analysis of the network by using CytoNCA plug-in Cytospace 3.2.1 software.With the help of DAVID database,the function of Gene Ontology(GO)was annotated and KEGG pathway was enriched;and the enrichment results were visualized through OmicShare Tools online mapping platform.RESULTS:Based on retrieval results of TCMSP database,44 active components were obtained,involving 13 of B.falcatum and 32 of S.baicalensis;stigmasterol is common component of B.falcatum and S.baicalensis.Quercetin,kaempferol and wogonin were the compounds with main potential targets.The target proteins with high node degree were prostaglandin endoperoxide synthase 2(PTGS2),PTGS1,nuclear receptor coactivator 2 and heat shock protein 90α,which were associated with 37,30,25 and 25 active components respectively.Twenty genes were obtained from the interaction between“B.falcatum-S.baicalensis”and podocyte lesion related targets,including PTGS2,VEGFA,MMP9,TNF and IL6.PPI network diagram of the above intersection genes contained 20 nodes and 110 lines,with MMP9,VEGFA,IL6 and other genes at the core.The results of GO analysis showed that a total of 154 biological information items were obtained(P<0.05),including 139 biological process items,8 cell composition items and 7 molecular function items.Among them,biological processes mainly involved in the positive regulation of NO biosynthesis process,inflammatory response,immune response.Cell composition mainly involved in extracellular space,extracellular region,external side of plasma membrane,etc.,and molecular function mainly involved in protein binding,cytokine activity,growth factor activity,etc.At the same time,47 KEGG pathways were obtained(P<0.05),mainly including cytokine-cytokine receptor interaction,rheumatoid arthritis,malaria,cancer signaling pathways.CONCLUSIONS:The active components of Compund ShensuⅡcouplet medicine of“B.falcatum-S.baicalensis”may act on MMP9,VEGFA,IL6,TNF and other targets through cytokine-cytokine receptor interaction,rheumatoid arthritis,malaria,cancer signal pathway,so as to play its intervention effect on podocyte lesion.