基于网络药理学和分子对接技术探讨益气开秘方治疗便秘的作用靶点及效应机制

Target and mechanism of Yiqi Kaimi Prescription in treatment of constipation based on network pharmacology and molecular docking

目的运用网络药理学和分子对接技术研究益气开秘方治疗便秘的作用靶点及效应机制。方法通过中药系统药理学数据库与分析平台收集筛选益气开秘方中组方药材的有效活性成分及对应靶点,利用GeneCards、OMIM、PharmGKB、TTD及DrugBank数据库获取疾病靶点;利用R软件4.1.1筛选成分和疾病的交集靶点,利用STRING数据库(version 11.5)和Cytoscape 3.8.2软件构建蛋白质–蛋白质相互作用(PPI)网络;运用R软件4.1.1进行基因本体(GO)功能及京都基因与基因组百科全书(KEGG)通路富集分析;运用Autodock软件对关键作用靶点和活性成分进行分子对接,验证网络分析结果。结果益气开秘方主要包含68个活性成分,活性成分对应靶点170个;便秘疾病靶点共1586个;益气开秘方与便秘的交集靶点基因共68个。PPI分析出核心靶点为丝裂原活化蛋白激酶1(mitogen-activated protein kinase 1,MAPK1)、MAPK3、蛋白激酶B1(AKT1)等;GO功能注释得到生物学过程条目1825条(如衰老、平滑肌细胞增殖、肽基-丝氨酸磷酸化等),细胞组成条目60条(如膜筏、膜微结构域、丝氨酸/苏氨酸蛋白激酶复合物等)、分子功能条目157条(如磷酸酶结合、内肽酶活力、丝氨酸肽链内切酶活性等);KEGG通路富集分析确定了168条益气开秘方治疗便秘作用靶点的通路(如PI3K/AKT信号通路、细胞衰老、MAPK信号通路、肌动蛋白细胞骨架调节等);分子对接提示MAPK1、MAPK3、AKT1与主要核心成分(槲皮苷、木犀草素以及山柰酚)的结合能均<-20 kJ/mol。结论益气开秘方中的槲皮苷、木犀草素以及山柰酚等活性成分可能通过与MAPK1、MAPK3、AKT1等靶点进而调控MAPK信号通路以及磷酸肌醇-3-激酶(PI3K)/AKT信号通路等信号通路起到治疗便秘的作用。

To study the target and mechanisms of Yiqi Kaimi Prescription in treatment of constipation based on network pharmacology and molecular docking.Methods The effective active components and corresponding targets of Yiqi Kaimi Prescription were collected and screened via TCMSP.The disease targets were obtained from the GeneCards,OMIM,PharmGKB,TTD and DrugBank databases.The intersection targets of components and diseases were screened by R software 4.1.1,and the protein interaction network was constructed by the STRING database(version 11.5)and Cytoscape 3.8.2.R software 4.1.1 was used for GO biological function analysis and KEGG pathway analysis.The key targets and active components were docked by Autodock software to verify the network analysis results.Results Yiqi Kaimi Prescription primarily contained 68 active ingredients,and the active ingredients corresponded to 170 targets.There were 1586 targets for constipation,and 68 target genes for the intersection of Yiqi Kaimi Prescription and constipation.PPI network analysis showed that the core targets were MAPK1,MAPK3 and AKT1.GO functional analysis showed 1825 biological processing items(such as aging,smooth muscle cell proliferation,peptide serine phosphorylation,etc.),60 cell composition items(such as membrane raft,membrane microdomain,serine/threonine protein kinase complex,etc.),and 157 molecular functional items(such as phosphatase binding,endopeptidase activity,serine endopeptidase activity,etc.).KEGG pathway enrichment analysis identified 168 pathways(such as PI3K/AKT signaling pathway,cell senescence,MAPK signaling pathway,actin cytoskeleton regulation,etc.)for the treatment of constipation;Molecular docking showed that the binding energies of MAPK1,MAPK3 and AKT1 with the central core components(quercetin,luteolin,and kaempferol)were<−20 kJ/mol.Conclusion Quercetin,luteolin and kaempferol in Yiqi Kaimi Prescription may regulate cellular aging,MAPK signaling pathway and PI3K/AKT signaling pathway by interacting with MAPK1,MAPK3 and AKT1.