基于网络药理学和分子对接技术探讨葛根素治疗早发性卵巢功能不全的作用机制

The Mechanism of Puerarin in the Treatment of Premature Ovarian Insufficiency Based on Network Pharmacology and Molecular Docking

目的:采用网络药理学及分子对接技术探究葛根素治疗早发性卵巢功能不全(premature ovarian insufficiency,POI)的作用机制。方法:通过中药系统药理学数据库与分析平台(traditional Chinese medicine systems pharmacology database and analysis platform,TCMSP)、STITCH数据库、SwissTargetPrediction数据库、Sea数据库和比较毒理基因组学数据库(comparative toxicogenomics database,CTD)检索葛根素的潜在作用靶点;利用在线人类孟德尔遗传数据库(online mendelian inheritance in man,OMIM)、人类基因数据库(the human gene database,GeneCards)和DisGeNet数据库检索获得早发性卵巢功能不全相关基因靶点;利用Cytoscape软件构建葛根素治疗POI的化合物-靶点-疾病网络,通过STRING平台构建蛋白-蛋白互作(protein-protein interactions,PPI)网络,并利用核心靶基因导入基因功能注释数据库(the database for annotation visualization and integrated discovery,DAVID)进行基因本体论(gene ontology,GO)及京都基因与基因组百科全书(kyoto encyclopedia of genesand genomes,KEGG)信号通路富集分析;利用AutoDock Vina以及PyMol对葛根素与核心靶点进行分子对接验证。结果:共获得176个葛根素潜在作用靶点,478个P0I相关基因;葛根素治疗P0I的潜在靶点共54个,核心靶点29个,包括Akt1、IL-6、VEGFA、EGF、MAPK1、IGF1、TNF、ESR1、TGFB1、ALB等靶点;GO和KEGG富集分析显示生物过程及信号通路主要集中于细胞增殖、炎症反应、免疫反应、对雌二醇的反应、生殖细胞发展、PI3K-Akt信号通路、FoxO信号通路、TGFβ信号通路、mTOR信号通路、雌激素信号通路、催乳素信号通路等;分子对接结果显示葛根素与靶蛋白具有较好结合活性及较稳定构象。结论:葛根素治疗POI具有多成分、多靶点、多通路特点。

Objective:To explore the mechanism of puerarin in the treatment of premature ovarian insufficiency(POI)using network pharmacology and molecular docking.Methods:The potential targets of puerarin were searched from TCMSP,STITCH database,SwissTargetPrediction database,Sea database and comparative toxicogenomics database(CTD);the relevant genetic targets of POI were retrieved and obtained using OMIM,GeneCards and DisGeNet database;Cytoscape software was used to construct the network of compound-targetdisease for the treatment of POI with puerarin,STRING platform was applied to construct protein-protein interactions(PPI)network,core target genes was imported into DAVID for GO and KEGG pathway enrichment analysis;AutoDockVina and PyMol were applied to conduct molecular docking validation of puerarin and core targets.Results:A total of 176 potential targets of puerarin and 478 POI-related genes were obtained;there were 54 potential targets and 29 core targets of puerarin in the treatment of POI,including Aktl,IL-6,VEGFA,EGF,MAPK1,IGF1,TNF,ESR1,TGFB1,ALB;GO and KEGG enrichment analysis displayed that the biological process and signaling pathway were mainly concentrated in cellular proliferation,inflammatory reaction,immunological reaction,the response to estradiol,reproductive cell development,PI3K-Akt signaling pathway,FoxO signaling pathway,TGFβ signaling pathway,mTOR signaling pathway,estrogen signaling pathway and prolactin signaling pathway;molecular docking results showed that puerarin has better binding activity and more stable conformation with target proteins.Conclusion:Puerarin has multi-ingredient,multi-target and multipathway properties in the treatment of POI.