基于网络药理学、分子对接和体外实验探讨熊果酸治疗脑胶质母细胞瘤的作用机制  

Exploring the Mechanism of Ursolic Acid in the Treatment of Glioblastoma Based on Network Pharmacology,Molecular Docking,and in Vitro Experiments

作　　者：高仁杰 崔瑞亭 颜凯 Gao Renjie;Cui Ruiting;Yan Kai(Taian City Central Hospital,Shandong First Medical University&Shandong Academy of Medical Sciences,Taian 271099,China;Intermal Medicine,People's Hospital of Sishui County,Sishui 273200,China)

机构地区：[1]山东第一医科大学(山东省医学科学院)泰安市中心医院,山东泰安271099 [2]山东省泗水县人民医院内科,山东泗水273200

出　　处：《南开大学学报（自然科学版）》2023年第4期16-24,共9页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：Supported by General Project of Taian Nursery Cultivation(2022MPM19)。

摘　　要：基于网络药理学、分子对接和体外实验探讨熊果酸(ursolic acid,UA)治疗脑胶质母细胞瘤(glioblastoma,GBM)的作用机制.通过TCMSP、SwissTargetPrediction和SuperPred数据库获取熊果酸活性作用靶点,同时采用GeneCards、DisGeNET、TTD和PharmGkb数据库结合两个GEO基因芯片库获取GBM疾病靶点,应用Cytoscape软件,筛选UA治疗GBM的核心靶点.通过GEPIA数据库分析核心靶点在GBM患者组织与正常组织中mRNA表达水平是否有显著差异.并采用David数据对潜在靶点进行基因本体(gene ontology,GO)功能及京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析.使用Autodock Vina和Pymol实现UA与重要的核心靶点分子对接.最后通过体外实验进一步验证了UA的抗肿瘤活性.结果表明,筛选出92个UA对应的靶点,获得7433个GBM靶点,合并后获得72个潜在靶点.PPI网络和拓扑分析结果表明,CCND1、EGFR、FOS、CASP3、HSP90AA1、IL6、CTNNB1、INS、RELA、VEGFA等10个核心靶标为核心靶点.GEPIA数据库分析显示,GBM组织中CASP3、CCND1、EGFR、FOS、IL-6和VEGFR mRNA的表达显著高于正常组织.GO富集分析发现,UA治疗GBM与细胞对化学应激的反应、膜筏、半胱氨酸型内肽酶活性等密切相关.KEGG通路分析表明,它涉及细胞凋亡、TNF信号通路、PI3K-Akt信号通路等.体外实验结果表明,UA可抑制U251细胞的增殖,IC50值为26.66μmol/L.RT-qPCR和WB的结果表明,UA可以调节U251细胞中CASP3和CCND1 mRNA及蛋白的表达,UA可促进U251细胞CASP3 mRNA和蛋白的过表达,并抑制CCND1 mRNA和蛋白表达.总之,UA可以通过多种靶点和途径治疗胶质母细胞瘤.Based on network pharmacology and molecular docking,combined with in vitro experiments,to explore the mechanism of ursolic acid(UA)in treating glioblastoma(GBM).UA targets were obtained from TCMSP,SwissTargetPrediction,SuperPred databases,and disease targets of GBM were obtained from four disease databases including GeneCards,DisGeNET,TTD,and PharmGkb combined with two GEO gene chips.Cytoscape software was used to screen the core target of UA in the treatment of GBM by protein interaction.GEPIA(gene expression profiling interactive analysis)database was used to analyze whether there is a significant difference in mRNA expression levels of core targets between GBM patient tissues and normal tissues.David database was used to carry out GO(gene ontology)function and KEGG(Kyoto encyclopedia of genes and genomes)pathway enrichment analysis for potential targets.Autodock Vina and Pymol were used to realize the molecular docking of UA and the significant core proteins.Finally,the anti-tumor activity of UA was further verified through in vitro experiments.The results indicated that 92 protein targets corresponding to UA were screened,and 7433 targets of GBM were obtained,and 72 potential targets were obtained after combination.The PPI network and topology analysis results show that 10 core targets,including CCND1,EGFR,FOS,CASP3,HSP90AA1,IL6,CTNNB1,INS,RELA,and VEGFA are the core targets.The expression of CASP3,CCND1,EGFR,FOS,IL-6,and VEGFR mRNA in GBM tissue was significantly higher than that in normal tissue by GEPIA database analysis.GO enrichment analysis found that UA treatment of GBM was closely related to cellular response to chemical stress,membrane raft,cysteine-type endopeptidase activity,etc.KEGG pathway analysis showed that it involved apoptosis,TNF signaling pathway,PI3K-Akt signaling pathway,etc.The results of in vitro experiments showed that UA could inhibit the proliferation of U251 cells,The IC50 value is 26.66μmol/L.The results of RT-qPCR and WB(western blot)showed that UA could regulate the expression

关 键 词：熊果酸 脑胶质母细胞瘤 网络药理学 分子对接 体外实验 机制 

分 类 号：R966[医药卫生—药理学]