The protect effect of equol and its potential targets in the context of diabetic nephropathy
Objective To study the potential role of equol in(diabetic nephropathy,DN)through network pharmacology methods,and validate the target for the protective effect of equol on DN.Methods The potential targets of Eq were identified through screening in databases such as PubChem,SwissTargetPrediction,and PharmMapper,intersecting with DN targets identified in DisGeNET,GeneCards,and OMIM.Subsequently,protein-protein interaction(PPI)analysis was conducted using the STRING platform.Enrichment analyses of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways were performed for key targets,and the"component target disease"network diagram was constructed by Cytoscape 3.8.0 software.Molecular docking was used to validate possible targets.Cultivate MPC5 cells in vitro and treat them with varying concentrations of Eq for 48 hours.The cell viability of each group was measured using the CCK-8 assay to determine the optimal concentration gradient for equol intervention.The final experi-mental group settings were as follows:Control group(GLU,5.5 mmol/L),Model group(GLU,30 mmol/L),LEq group(model+1×10-8mol/L Eq),MEq group(model+1×10-7mol/L Eq),and HEq group(model+1×10-6mol/L Eq).Apoptosis in each group was analyzed using flow cytometry.Western bloting was used to detect the protein expres-sion levels of EGFR,P-EGFR,Bcl-2,Bax and Cleaved Caspase-3 in each group.Results A total of 128 intersection targets of Eq and DN were obtained.The core targets were PIK3CB、PIK3CA、AKT2、MAPK1、HRAS、RAF1、MAP2K1 and EGFR.The findings from the flow cytometry experiments indicated that,in comparison to the model group,the cell survival rate and apoptosis rate were notably elevated following Eq intervention.Western bloting experiment indicated that the expression level of anti-apoptotic protein Bcl-2 was significantly higher than that of the model group after equol intervention(P<0.05).The protein expression levels of P-EGFR/EGFR,Bax and Cleaved Caspase-3 were significantly reduced(P<0.05).Conclusion Equol exhibits multi-target and pathway regulatory effects against DN.Intervention with equol can alleviate the phenomenon of apoptosis in renal podocytes,and its mechanism may be related to the EGFR signaling pathway.
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