基于网络药理学和分子对接探讨珠子参地上部分防治脂肪肝的作用机制

Mechanism of aerial part of Rhizoma Panacis Majoris for fatty liver based on network pharmacology and molecular docking

目的探讨珠子参地上部分防治脂肪肝的活性成分、作用靶点、分子通路,揭示珠子参地上部分防治脂肪肝的作用机制,并初步验证可能的作用机制。方法通过TCMSP和Swiss TargetPrediction数据库以及文献挖掘获取珠子参地上部分活性成分与预测靶点;然后检索Genecards数据库得到脂肪肝相关的疾病靶点;将预测靶点与疾病靶点取交集,筛选出共同的脂肪肝靶点即关键靶点,构建"药物-成分-疾病-靶点"网络;采用String数据库构建活性成分和关键靶点的蛋白相互作用网络(PPI),通过R project软件对关键靶点进行GO富集分析和KEGG通路富集分析;利用Autodock_vina软件进行活性成分和核心靶点的分子对接验证;测定三酰甘油(TG)、总胆固醇(TC)的水平,并通过免疫组化法验证珠子参地上部分对环氧合酶2(PTGS2)、丝裂原激活的蛋白激酶3(MAPK3)、过氧化物酶体增殖物激活受体γ(PPARG)的影响。结果最终筛选出珠子参地上部分的活性成分5个,靶点175个;脂肪肝相关的疾病靶点1481个;构建关键作用靶点的PPI网络,其中包含节点84个,边513条;分子对接结果显示PTGS2、MAPK3、PPARG 3个核心靶点能够与活性成分结合并借助氢键等分子间作用力形成较为稳定的构象;GO富集分析主要涉及的生物学功能与过程有类固醇代谢过程、脂肪酸代谢过程、不饱和脂肪酸代谢过程、脂肪酸衍生物代谢过程等,KEGG通路富集分析显示珠子参地上部分通过PPAR信号通路、非酒精性脂肪肝通路、细胞色素P450对外源生物的代谢通路、脂肪消化吸收通路等来发挥防治脂肪肝的作用;免疫组化实验证明珠子参地上部分能够抑制小鼠肝脏组织中PTGS2蛋白的表达,增强MAPK3和PPARG蛋白的表达。结论本研究通过网络药理学,结合分子对接和免疫组化验证,发现珠子参地上部分可能通过多成分、多靶点、多途径协同发挥防治脂肪肝的作用,可为珠子参地上部分的应用提供新的理论基础。

Objective To determine the active aerial part of Rhizoma Panacis Majoris(APRPM)mechanism in the prevention and treatment of fatty liver,and verify its mechanism.Methods Active components and prediction targets of APRPM were obtained by TCMSP and Swiss TargetPrediction.Then the disease targets related to fatty liver were obtained by searching Genecards database.The common fatty liver targets were selected by intersecting the prediction targets and disease targets,and the"drug-component-disease-target"network was constructed and its action mechanism was preliminarily analyzed.The protein-protein interaction network of the key targets was constructed with String database to analyze the GO enrichment and KEGG pathway enrichment of the key targets by R project software.The molecular docking verification of active components and core targets was conductecd with Autodock_vina software.Finally,The levels of triglyceride(TG),and total cholesterol(TC)were determined and the effects of APRPM on cyclooxygenase 2(PTGS2),mitogen-activated protein kinase 3(MAPK3)and peroxisome proliferator-activated receptorγ(PPARG)verified by immunohistochemistry.Results Five active components and 175 targets of APRPM were screened out,1481 disease targets related to fatty liver were selected and the PPI network of key targets was constructed,including 84 nodes and 513 edges.Molecular docking showed that the 3 core targets of PTGS2,MAPK3 and PPARG combined with the active components and formed a relatively stable conformation with the help of intermolecular forces such as hydrogen bonds.The main biological functions and processes involved in the GO enrichment analysis included steroid metabolism,fatty acid metabolism,unsaturated fatty acid metabolism,fatty acid derivative metabolism and so on.KEGG pathway enrichment analysis showed that PPAR signal pathway,non-alcoholic fatty liver disease,metabolism of xenobiotics by cytochrome P450,and fat digestion and absorption all played a role in the prevention and treatment of fatty liver.Immunohistochemical experiment showed that APRPM inhibited the expression of PTGS2 protein and enhanced the expression of MAPK3 and PPARG protein in the mouse liver.Conclusion Network pharmacology,molecular docking and immunohistochemistry show that APRPM prevent and treat fatty liver through the synergistic action of multi-components,multi-targets and multi-pathways.This paper provides a new theoretical basis for the application of APRPM.