基于网络药理学与分子对接技术研究淫羊藿治疗少弱精症的作用机制  

Mechanism of Epimedii Folium in Treatment of Oligoasthenotspermia Based on Network Pharmacology and Molecular Docking

作　　者：方春燕 叶玉龙 叶乃菁 申毅锋 俞旭君[1] 常德贵[1] FANG Chunyan;YE Yulong;YE Naijing;SHEN Yifeng;YU Xujun;CHANG Degui(Traditional Chinese Medicine(TCM)Regulating Metabolic Diseases Key Laboratory of Sichuan Province,Hospital of Chengdu University of TCM,Chengdu 610075,China;Tea Research Institute,Sichuan Academy of Agricultural Sciences,Chengdu 610066,China)

机构地区：[1]成都中医药大学附属医院,代谢性疾病中医药调控四川省重点实验室,成都610075 [2]四川省农业科学院茶叶研究所,成都610066

出　　处：《中国实验方剂学杂志》2022年第19期194-203,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年基金项目(82104830);国家自然科学基金面上项目(81973647)。

摘　　要：目的:利用网络药理学、分子对接技术探究淫羊藿治疗少弱精症的有效成分及作用机制。方法:利用中药系统药理学数据库与分析平台(TCMSP)筛选出淫羊藿主要活性成分及对应靶点基因;通过GeneCards、在线人类孟德尔遗传数据库(OMIM)获得少弱精症靶点基因;利用UniProt对所有基因进行校正;利用Cytoscape 3.9.0软件构建药物-活性成分-关键靶点调控网络,根据度值筛选关键活性成分;将活性成分与疾病共同靶点上传至STRING 11.5数据库构建淫羊藿靶蛋白-少弱精症靶蛋白质-蛋白质相互作用(PPI)网络,根据度值筛选关键蛋白靶点;利用DAVID数据库对关键靶点进行基因本体(GO)功能富集分析及京都基因与基因组百科(KEGG)通路富集分析;利用Protein Data Bank(PDB)数据库及TCMSP数据库获得靶蛋白和活性成分的分子结构,通过AutoDock Vina 1.1.2软件对活性成分及核心蛋白靶点进行分子对接;最后通过淫羊藿活性成分淫羊藿苷干预少弱精症模型大鼠,验证淫羊藿苷对蛋白靶点表达的影响。结果:从淫羊藿中筛选出23个活性成分,淫羊藿与少弱精症的共同关键靶点50个,核心靶点6个,包括肿瘤蛋白p53(TP53)、表皮生长因子受体(EGFR)、前列腺素G/H合酶2(PTGS2)、胱天蛋白酶(Caspase)-3、受体酪氨酸蛋白激酶erbB-2(ERBB2)、Caspase-9。GO功能富集与KEGG信号通路富集分析淫羊藿活性成分主要参与P53信号通路、癌症途径、细胞增殖和凋亡等。分子对接结果提示淫羊藿苷、槲皮素、8-异戊烯醇三个活性成分与靶蛋白结合能力较强。淫羊藿苷干预实验结果表明,与空白组比较,少弱精模型大鼠睾丸中靶蛋白表达显著下调;与少弱精症模型大鼠组比,淫羊藿苷可显著上调靶蛋白在淫羊藿苷组睾丸中靶蛋白的表达(P<0.05)。结论:淫羊藿主要通过淫羊藿苷、8(-3-甲基丁-2-烯基)-2-苯基色酮、8-异戊烯醇,调节P53信号通路、癌症途径、细Objective:To explore the effective components and mechanism of Epimedii Folium in the treatment of oligoasthenotspermia by using network pharmacology and molecular docking technique.Method:The main active components and corresponding target genes of Epimedii Folium were screened out from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP).Target genes of oligospermia were obtained by GeneCards and Online Mendelian Inheritance in Man(OMIM)database.Uniprot was used to correct all genes.The drug-active component-key target regulatory network was constructed by Cytoscape3.9.0,and the key active components were screened out according to the degree value.The active components and common targets of the disease were uploaded to STRING 11.5 database to construct the Epimedii Folium and oligoasthenotspermia target protein-protein interaction(PPI)network,and the key protein targets were screened out according to the degree value.The key targets of gene ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed using DAVID database.Protein Data Bank(PDB)and TCMSP were used to obtain the molecular structure of target proteins and active components.AutoDock Vina 1.1.2 was used to perform molecular docking of the active components and the core protein targets.Finally,icariin,the active component of Epimedii Folium,was used to intervene in the rat model of oligoasthenotspermia to verify the effect of icariin on the expression level of protein targets.Result:Twenty-three active components from Epimedii Folium were screened out,and 50 common targets and 6 core targets of oligoasthenotspermia and Epimedii Folium were obtained,including tumor protein p53(TP53),epidermal growth factor receptor(EGFR),prostaglandinendoperoxide synthase 2(PTGS2),cysteine aspartate-specific protease(Caspase)-3,erb-b2 receptor tyrosine kinase 2(ERBB2),and caspase-9.Through GO enrichment and KEGG pathway enrichment analysis,the active components

关 键 词：淫羊藿 少弱精症 网络药理学 分子对接 实验验证 

分 类 号：R285[医药卫生—中药学] R289[医药卫生—中医学] R22R2-031R33