基于生物信息学和实验验证分析六味地黄汤治疗肾纤维化的分子机制  

Molecular Mechanism of Liuwei Dihuang Decoction(六味地黄汤)in the Treatment of Renal Fibrosis Based on Bioinformatics and Experimental Verification

作　　者：杨思禹 张海英 郭金晶 孟斌[1] 刘春燕[1] 唐群[1] YANG Siyu;ZHANG Haiying;GUO Jinjing;MENG Bin;LIU Chunyan;TANG Qun(Hunan University of Chinese Medicine,Changsha 410208)

机构地区：[1]湖南中医药大学,长沙410208

出　　处：《中药药理与临床》2024年第3期29-35,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：湖南省教育厅项目(编号:21A0242、20C1398);湖南省自然科学基金项目(编号:2021JJ30506);长沙市自然科学基金项目(编号:kq2014089);湖南中医药大学校级科研基金(编号:2020-27)。

摘　　要：目的:基于生物信息学分析六味地黄汤治疗肾纤维化的分子机制并验证。方法:从美国国家生物技术信息中心(NCBI)旗下的基因表达数据库(GEO)获得包含29个正常肾组织和23个肾纤维化组织的基因芯片数据集GSE12682和注释文本GPL571,使用Strawberry Perl和R软件筛选肾纤维化差异表达基因(DEGs)。通过中药系统药理学数据库与分析平台(TCMSP)获得六味地黄汤中六味药的成分表和作用基因靶点。使用Strawberry Perl整合出两者的交叉靶点,并用Cytoscape软件和R软件制作蛋白质-蛋白质相互作用(Protein-protein interaction,PPI)网络,利用STRING网站进行基因本体(GO)生物功能分析、京都基因与基因组百科全书(KEGG)通路富集分析。12只SD雄性大鼠随机分为正常对照组、模型对照组、依那普利10 g/kg组和六味地黄汤6.25 g/kg组,HE染色、Masson染色观察肾组织病理改变,免疫组化法检测肾组织血管内皮生长因子信号A(VEGFA)、血管内皮生长因子信号受体1(VEGFR1)两种蛋白的表达差异。结果:通过疾病、药物取交集靶点得到六味地黄汤针对肾纤维化的26个基因靶点;GO提示这些基因影响2种细胞组分、210条生物过程;KGEE表明关键通路为血管内皮生长因子信号(VEGF)信号通路、ERBB信号通路、FcεRI信号通路、白介素-17(IL-17)信号通路等。验证试验表明,与正常对照组相比,模型对照组肾组织胶原容积分数增加,VEGFA和VEGFR1蛋白表达明显下调(P<0.05);与模型对照组相比,六味地黄汤组肾组织胶原容积分数降低,VEGFA和VEGFR1蛋白表达明显上调(P<0.05)。结论:六味地黄汤可通过多基因靶点、多信号通路治疗肾纤维化,其中上调VEGF信号通路是其抗纤维化的重要分子机制。Objective:To analyze and validate the molecular mechanism of Liuwei Dihuang Decoction(六味地黄汤)in the treatment of renal fibrosis based on bioinformatics.Methods:The gene chip data set GSE12682 and annotation text GP1571 were obtained from Gene Expression Omnibus(GEO)database under the National Center for Biotechnology Information(NCBI)in the US,including 29 normal renal tissues and 23 renal fibrotic tissues.Strawberry Perl and R software were used to screen differentially expressed genes(DEGs)of renal fibrosis.Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)was used to obtain the components and gene targets of six drugs in Liuwei Dihuang Decoction.Strawberry Perl was used to integrate the common targets,and Cytoscape software and R software were used to plot the protein-protein interaction(PPI)network.STRING was used for Gene Ontology(GO)biological function analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.In experimental verification,12 male SD rats were randomly divided into a blank control group,a model group,an enalapril group,and a Liuwei Dihuang Decoction group.HE staining and Masson staining were used to observe the pathological changes of renal tissues in each group,and the immunohistochemical method was used to detect the protein expression of vascular endothelial growth factor A(VEGFA)and vascular endothelial growth factor receptor 1(VEGFR1)in renal tissues in each group.Result:Twenty-six gene targets of Liuwei Dihuang Decoction on renal fibrosis were obtained by intersecting targets of the disease and drugs.GO suggested that these genes affected two cellular components and 210 biological processes.KCEE showed that the key pathways were the VEGF signaling pathway,ERBB signaling pathway,FCeRI signaling pathway,IL-17 signaling pathway,etc.As revealed by experimental verification,the collagen volume fraction of the model group increased and the protein expression levels of VECFA and VECFR1 were down-regulated as compared wit

关 键 词：六味地黄汤 生物信息学 肾纤维化 分子机制 血管内皮生长因子信号通路 

分 类 号：R285[医药卫生—中药学]