The Mechanism Research of Tangshenbao Compound in Treating Diabetic Nephropathy Based on Network Pharmacology and Experimental Verification
The research investigated the mechanism of action of Tangshenbao compound(TSB)in treating diabetic nephropathy(DN)by combining network pharmacology methods with molecular biology experiments.The active ingredients and targets relat-ed to TSB were obtained using TCMSP,Pubchem,and Swiss Target Prediction databases,along with relevant literature.Candi-date targets for TSB in treating DN were obtained through GeneCards,DisGeNET,and OMIM databases.A Venn diagram of the"TSB-DN"candidate targets was drawn using online plotting software,followed by GO and KEGG enrichment analysis using Cytoscape 3.7.2 software.Subsequently,molecular docking of active ingredient ligands with target protein crystal structures was completed using PyMOL,AutoDock Tools,and AutoDock Vina software.Cell experiments were then conducted using Mcs me-sangial cells cultured in vitro to validate the findings.Real-time fluorescence quantitative PCR was used to detect the mRNA ex-pression levels of STAT3,PIK3CD,PIK3R1,MAPK8,and their associated targets in the cells of each group,while cell viabili-ty after 24 hours was assessed using the CCK-8 method.Results showed that a total of 835 TSB target genes were obtained after processing the Swiss Target Prediction database.When combined with GeneCards,DisGeNET,OMIM,and other disease data-bases,there were 443 intersecting target genes for"TSB-DN".Protein-protein interaction(PPI)network analysis identified the top 10 degree targets as SRC,PIK3R1,STAT3 and PIK3CA.GO and KEGG enrichment analysis results indicated that TSB treatment for DN may be related to functions such as protein phosphorylation,cell signal transduction,and positive regulation of gene expression.Finally,cell experiments in vitro demonstrated that TSB could effectively reduce the mRNA expression levels of PIK3CD,PIK3R1,and ETS-1 in SV40-MES-13 cells,resulting in an"inhibitory,sedative"effect following drug action on the cells.The results indicated that the TSB compound can inhibit the mRNA expression of target genes such as PIK3R1,PIK3CD,MAPK8,STAT3,and ETS-1,thereby regulating the PI3K-Akt and AGE-RAGE signaling pathways and exerting a therapeutic effect on DN.
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维普
