基于网络药理学和分子对接技术探讨贞芪扶正颗粒辅助治疗肺结核的作用机制

Mechanism of Zhenqi Fuzheng Granules for the adjuvant treatment of pulmonary tuberculosis:an exploration based on network pharmacology and molecular docking technique

目的基于网络药理学和分子对接技术探讨贞芪扶正颗粒辅助治疗肺结核的作用机制。方法利用中药系统药理学数据库与分析平台检索贞芪扶正颗粒的活性化学成分及其作用靶点,利用GeneCards数据库检索肺结核相关靶点,取两者交集靶点。采用Cytoscape 3.7.2软件构建“中药-成分-靶点”网络以获取主要活性化学成分。采用STRING数据库及Cytoscape 3.7.2软件构建交集靶点的蛋白-蛋白相互作用网络,筛选核心靶点。对核心靶点进行基因本体论(GO)功能富集分析及京都基因与基因组百科全书(KEGG)通路富集分析。使用AutoDock软件对主要活性化学成分和核心靶点进行分子对接。结果共得到贞芪扶正颗粒的活性化学成分31个及其作用靶点198个,肺结核相关靶点2415个,交集靶点101个。主要活性化学成分包括槲皮素、木犀草素、山柰酚、β-谷甾醇、芒柄花素;核心靶点包括白细胞介素(IL)-6、丝氨酸/苏氨酸蛋白激酶1、IL-1β、肿瘤蛋白p53、基质金属蛋白酶9(MMP9)、Caspase-3、原癌基因c-Jun、C-X-C基序趋化因子配体8(CXCL8)、前列腺素内过氧化物合酶2(PTGS2)、过氧化物酶体增生激活受体γ(PPARG)。GO功能富集分析结果显示,核心靶点涉及的生物学过程主要有机械刺激反应、生物调节、代谢过程等,涉及的细胞组分主要有细胞膜、细胞核、膜封闭管腔等,涉及的分子功能主要包括蛋白质结合、核苷酸结合、离子结合等。KEGG通路富集分析结果显示,核心靶点涉及的信号通路主要包括IL-17信号通路、肿瘤坏死因子信号通路等。分子对接结果显示,MMP9、IL-6、PTGS2、PPARG、CXCL8与贞芪扶正颗粒的主要活性化学成分均有稳定的对接效果(结合能<-5.0 kcal/mol)。结论贞芪扶正颗粒辅助治疗肺结核的作用机制具有多成分、多靶点和多通路的特点,其活性化学成分槲皮素、木犀草素等可能通过作用于MMP9、IL-6、PTGS2、PPARG、CXCL8等靶点,来抑制IL-7信号通路、肿瘤坏死因子信号通路,从而发挥辅助治疗肺结核的作用。

Objective To explore the mechanism of Zhenqi Fuzheng Granules for the adjuvant treatment of pulmonary tuberculosis based on network pharmacology and molecular docking technique.Methods The active chemical components and their effect targets of Zhenqi Fuzheng Granules were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,and the targets related to pulmonary tuberculosis were retrieved from the GeneCards database;furthermore,the intersection targets of the two were obtained.The"Traditional Chinese Medicine-component-target"network was constructed by using the Cytoscape 3.7.2 software to obtain main active chemical components.The protein-protein interaction network of the intersection targets was constructed to screen core targets by employing the STRING database and Cytoscape 3.7.2 software.The Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed on the core targets.The AutoDock software was used for molecular docking of the main active chemical components and core targets.Results A total of 31 active chemical components and 198 effect targets of Zhenqi Fuzheng Granules were obtained,and there were 2415 targets related to pulmonary tuberculosis,as well as 101 intersection targets.The main active chemical components contained quercetin,luteolin,kaempferol,β-sitosterol,and formononetin;in addition,the core targets included interleukin(IL)-6,serine/threonine protein kinase 1,IL-1β,tumor protein p53,matrix metalloproteinase 9(MMP9),Caspase-3,proto-oncogene c-Jun,C-X-C motif chemokine ligand 8(CXCL8),prostaglandin-endoperoxide synthase 2(PTGS2),and peroxisome proliferative activated receptor gamma(PPARG).The results of GO functional enrichment analysis revealed that the biological processes involved in the core targets were mainly mechanical stimulation response,biological regulation,metabolic process,etc.,the cell compositions involved in were mainly cell membrane,cell nucleus,membrane closed lumen,etc.,and the molecular function involved in were mainly protein binding,nucleotide binding,ion binding,etc.The results of KEGG pathway enrichment analysis indicated that the signaling pathway involved in the core targets mainly contained IL-17 signaling pathway and tumor necrosis factor signaling pathway,etc.The molecular docking results depicted that MMP9,IL-6,PTGS2,PPARG,CXCL8 had stable docking effects with main active chemical components of Zhenqi Fuzheng Granules(with binding energy<-5.0 kcal/mol).Conclusion The mechanism of Zhenqi Fuzheng Granules for the adjuvant treatment of pulmonary tuberculosis has the characteristics of multi-components,multi-targets and multi-pathways,its active chemical components with respect to quercetin,luteolin,and other active chemical components may inhibit IL-7 signaling pathway and tumor necrosis factor signaling pathway,so as to exert its adjuvant therapeutic effect on pulmonary tuberculosis through acting on MMP9,IL-6,PTGS2,PPARG,CXCL8,and other targets.