摘要
目的 探讨黄芪抑制胶质瘤的作用机制和基因靶点.方法 基于网络药理学分析获得黄芪可能治疗胶质瘤的基因靶点,采用基因本体论(GO)富集分析和京都基因和基因组数据库(KEGG)通路分析,构建蛋白质-蛋白质相互作用(PPI)网络并筛选出Hub基因,最后构建疾病-药物-成分-靶点-通路的可视化网络.同时取胶质瘤细胞株U251进行细胞实验论证,采用细胞计数试剂盒、活死细胞双染色法检测不同浓度黄芪培养液对U251细胞存活率和凋亡的影响.结果 共筛选出145个黄芪可能治疗胶质瘤的基因靶点,GO富集分析显示最主要的生物学功能包括不同蛋白结合、相同蛋白结合、RNA聚合酶Ⅱ启动子转录的正向调控、酶结合等,KEGG通路分析显示最主要的信号通路包括癌症通路、脂质与动脉粥样硬化、流体剪切应力和动脉粥样硬化、化学致癌-受体激活等.Hub基因关联程度从高到低依次为血管内皮生长因子A、蛋白激酶B(AKT)1、JUN、基质金属蛋白酶9、PTGS2、IL-6、IL-1B、CASP3、TP53、HIF1A.在所有生物活性成分中,MOL000098(槲皮素)的度值最高,为118;在所有信号通路中,hsa05200(癌症通路)的度值最高,为61;在所有基因靶点中,AKT1、RELA、PTGS2、MAPL1、TP53的度值较高,分别为21、20、20、19、18.细胞实验证实,不同浓度(1、5、10mg/mL)黄芪培养液对U251细胞增殖均有抑制和促进凋亡作用,其作用强度呈浓度依赖性(均P＜0.05).结论 黄芪对U251细胞增殖具有抑制和促进凋亡作用,主要基因靶点包括AKT1、RELA、PTGS2、MAPL1、TP53.
Abstract
Objective To investigate the mechanisms and genetic targets through which Astragalus membranaceus inhibits gliomas.Methods Based on network pharmacology analysis,the potential gene targets of Astragalus membranaceus for glioma were acquired.The gene ontology(GO)enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)pathway analysis were applied to construct a protein-protein interaction(PPI)network and identify Hub genes,ultimately creating a visual network encompassing disease-drug-components-targets-pathways.Concurrently,U251 glioma cell lines were subjected to cellular experiments for validation.The impact of Astragalus membranaceus culture fluid at varying concentrations on U251 cell viability and apoptosis was assessed using a cell counting kit and live/dead cells dual staining method.Results A total of 145 potential gene targets of Astragalus membranaceus for the treatment of gliomas were identified.GO enrichment analysis revealed the predominant biological functions,including binding of different proteins,binding of identical proteins,positive regulation of transcription from RNA polymerase Ⅱ promoter,enzyme binding,etc.Meanwhile,KEGG pathway analysis revealed the principal signaling pathways,encompassing pathways in cancer,lipid and atherosclerosis,fluid shear stress and atherosclerosis,chemical carcinogenesis-receptor activation,etc.The Hub genes were ranked in descending order of association intensity as VEGFA,protein kinase B(AKT)1,JUN,MMP9,PTGS2,IL-6,IL-1B,CASP3,TP53,and HIF1A.Among all bioactive constituents,MOL000098(quercetin)exhibited the highest centrality,with a degree value of 118.Within the spectrum of signaling pathways,hsa05200(cancer pathway)attained the highest degree value,reaching 61.Amongst all gene targets,AKT1,RELA,PTGS2,MAPL1 and TP53 demonstrated elevated centrality,with degree values of 21,20,20,19 and 18,respectively.Cellular experiments validated that Astragalus membranaceus culture fluid at different concentrations(1,5,10 mg/mL)exerted inhibitory and apoptotic effects on U251 cell proliferation,with the intensity of its effects exhibiting concentration dependency(all P＜0.05).Conclusion Astragalus membranaceus demonstrates inhibitory and apoptotic effects on U251 cell proliferation,with primary gene targets including AKT1,RELA,PTGS2,MAPL1,and TP53.
维普
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