基于网络药理学和分子对接探索桂枝茯苓丸治疗卵巢癌的作用机制

Study on the mechanism of Guizhi Fuling pill in the treatment of ovarian cancer based on network pharmacology and molecular docking

目的:基于网络药理学和分子对接技术探索桂枝茯苓丸治疗卵巢癌的主要活性成分和作用机制。方法:使用TCMSP数据库检索并筛选桂枝茯苓丸中每味中药的活性成分,在Uniprot数据库找出药物活性成分的作用靶点,通过Cytoscape软件的Network Analyzer功能筛选出药物的主要活性成分及靶点。使用Genecards、OMIM、Digsee、TTD、Drugbank数据库检索卵巢癌的相关靶点。用R软件对药物主要活性成分的作用靶点和卵巢癌相关靶点取交集。将交集靶点导入String数据库中进行PPI分析,再把结果导入到Cytoscape中使用Cytohubba插件得到枢纽基因。在Metascape数据库中对枢纽基因进行KEGG通路分析和GO富集分析。最后使用Auto Dock Tools将得到的药物主要活性成分和筛选出的靶蛋白进行分子对接验证,Py Mol和Lig Plus软件将结果可视化。结果:得到了8个桂枝茯苓丸主要活性成分,分别为槲皮素、β-谷甾醇、山奈酚、常春藤皂苷元、儿茶素、鞣花酸、豆甾醇、花旗松素。药物对卵巢癌作用的枢纽基因20个,分别为VEGFA、AKT1、MAPK8、JUN、MMP9、IL-6、TNF、CXCL8、PTGS2、TP53、CASP3、MAPK1、EGF、ESR1、EGFR、MYC、FOS、CCL2、CXCL8和IL-1B。KEGG通路分析得到相关性最高的10条通路,分别为癌症信号通路、MAPK信号通路、流体剪切应力与动脉粥样硬化、乙型肝炎、美国锥虫病、糖尿病并发症中的年龄-年龄信号通路、肿瘤坏死因子信号通路、百日咳、IL-17信号通路、膀胱癌。GO富集分析在分子生物学功能、细胞组分、生物过程方面各筛选出了重要性较高的10个条目。IL-6、MAPK1、VEGFA、AKT1、TP53、TNF和CCL2编码的蛋白作为靶点分别与药物主要活性成分进行对接,对接结果良好,最好的是TNF-α和鞣花酸。结论:桂枝茯苓丸的主要活性成分通过调节IL-6、VEGFA、CCL2、TNF、MMP9等多种细胞因子调控多条信号通路,抑制癌细胞的增殖和转移。

Objective:To explore the main active components and mechanism of Guizhi Fuling pill in the treatment of ovarian cancer based on network pharmacology and molecular docking technology.Methods:TCMSP database was used to retrieve and screen the active components of each Chinese medicine in Guizhi Fuling pill.The main active components and targets of the medicine were screened through the Network Analyzer function of Cytoscape software,and the target of the main active components of the medicine was found in Uni Prot database.Genecards,OMIM,Digsee,TTD,and Drugbank database were used to retrieve the related targets of ovarian cancer.The interaction between the target of the main active components of the drug and the related target of ovarian cancer was obtained by R software.The intersection targets were imported into String database for PPI analysis,and then the results were imported into Cytoscape to get the hub gene using the Cyclohubba plug-in.KEGG pathway analysis and GO enrichment analysis were carried out in Metascape database.At last,Auto Dock Tools was used to verify the main active ingredients and the selected target proteins.Py MOL and Lig Plus software visualized the results.Results:The main active components of Guizhi Fuling pills were quercetin,β-sitosterol,kaempferol,Ivy saponin,catechin,ellagic acid,stigmasterol,and Douglas fir.There were 20 pivotal genes of drug effect on ovarian cancer,including VEGFA,AKT1,MAPK8,JUN,MMP9,IL-6,TNF,CXCL8,PTGS2,TP53,CASP3,MAPK1,EGF,ESR1,EGFR,MYC,FOS,CCL2,CXCL8 and IL-1 B.According to KEGG pathway analysis,10 pathways with the highest correlation were found,including cancer signaling pathway,MAPK signaling pathway,fluid shear stress and atherosclerosis,hepatitis B,trypanosomiasis,age-rage signaling pathway in diabetic complications,tumor necrosis factor signaling pathway,pertussis,IL-17 signaling pathway,and bladder cancer.GO enrichment analysis screened out 10 important items in molecular biological function,cell components and biological process.The proteins encoded by IL-6,MAPK1,VEGFA,AKT1,TP53,TNF and CCL2 were used as targets to dock with the main active components of the drug respectively,and the best results were TNF-α and ellagic acid.Conclusion:The main active components of Guizhi Fuling pill regulate multiple signal pathways by regulating IL-6,VEGFA,CCL2,TNF,MMP9 and other cytokines,and inhibit the proliferation and metastasis of cancer cells.