Exploring The Renal Protective Effect And Mechanism Of Zhengxintai Granules On Diabetic Nephropathy Rat Model Based On AGEs-RAGE Pathway

Purpose: To investigate the pharmacological effects and potential molecular mechanisms of Zhengxintai granules in the treatment of diabetic nephropathy using bioinformatics and in vivo experiments.Methods: A rat model of diabetic nephropathy was constructed by high sugar and high fat diet + intraperitoneal injection of streptozotocin,and Zhengxintai granules were gavaged,during which fasting blood glucose of the rats were measured,and the rats were executed after 3 weeks,and the renal index,24 h urine protein,biochemical indexes and histopathological analysis were detected.Bioinformatics analysis of Zhengxintai granules for the treatment of diabetic nephropathy.The chemical composition and corresponding targets of six Chinese herbal medicines in Zhengxintai granules,namely Astragalus,Pueraria Mirifica,Salvia,Fructus Crataegi,Rhizoma Chuanxiong and Quercus,were obtained from the Chinese medicine database,and the key data sets of diabetic nephropathy were obtained in the GEO database,and the differential genes and gene coexpression network,i.e.disease targets,were obtained after R language processing,and the intersection targets of the three As the potential targets of Zhengxintai granules for the treatment of diabetic nephropathy,we used String data to construct protein interactions network,Cyto Hubba plug-in in Cytoscape software to obtain core targets and pivotal genes,and constructed Chinese herbal medicine-active ingredient-target-disease network diagram by Cytoscape software,and the substances ranked in the top three according to Degree value were the core substances of Zhengxintai granules.The core targets were enriched and analyzed,and the pivotal genes were molecularly docked with the main components.Based on the bioinformatics prediction results,the AGEs-RAGE signaling pathway and its related pathways were analyzed to the SOD,MDA,PI3 K,p-AKT,NF-κBp65,NF-κBp-p65,IL-1β,CXCL8,CCL2,BAX,BCL2 were detected by ELISA kit and WB expression.Results: Zhengxintai granules improved blood glucose,body weight,urea nitrogen,urine creatinine,24 h urine volume,24 h urine protein,renal index and renal histopathological changes in rats with diabetic nephropathy.This study obtained a total of 18 core targets of Zhengxintai granules.Zhengxintai granules for the treatment of diabetic nephropathy,among which the pivotal genes include: CXCL2,CCL2,Caspase-3,Main components involved: quercetin,kaempferol,dousterol.KEGG indicates that Zhengxintai granules for the treatment of diabetic nephropathy mainly involves AGEs-RAGE signaling pathway,molecular docking results show that Zhengxintai of the molecular docking results showed that the main components of Zhengxintai pellets bound to the pivotal gene with <-5kcal/mol,showing good binding activity.The oxidative indexes MDA and SOD in rat kidney were improved to different degrees by the kit,and the enzyme immunoassay kit showed that Zhengxintai granules could improve the expression of AGEs,RAGE,CXCL2,CCL2,IL-1β,Caspase-3,BAX and BCL2 in the kidney and serum of rats with diabetic nephropathy.WB results showed that Zhengxintai granules could activate the expression of PI3K-AKT signaling pathway and inhibit the expression of NF-κBp65 to exert antiapoptotic and anti-inflammatory effects.Conclusion: Through bioinformatics and animal experiments,Zhengxintai granules exerted anti-apoptotic and anti-inflammatory effects through AGEs-RAGE signaling pathway to reduce kidney damage in diabetic nephropathy.