Molecular Mechanism of Liuwei Dihuang Decoction(六味地黄汤)in the Treatment of Renal Fibrosis Based on Bioinformatics and Experimental Verification
Objective:To analyze and validate the molecular mechanism of Liuwei Dihuang Decoction(六味地黄汤)in the treatment of renal fi-brosis based on bioinformatics.Methods:The gene chip data set GSE12682 and annotation text GPl571 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.Tradi-tional 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 ran-domly 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.KGEE 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 VEGFA and VEG-FR1 were down-regulated as compared with those in the blank control group(P＜0.05).Compared with the results in the model group,the collagen volume fraction of Liuwei Dihuang Decoction group decreased and the protein expression levels of VEGFA and VEGFR1 were up-reg-ulated(P＜0.05).Conclusion:Liuwei Dihuang Decoction can treat renal fibrosis through multiple gene targets and multiple signaling path-ways,and the up-regulation of the VEGF signaling pathway is an important molecular mechanism of fibrosis resistance.
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