关键词:

"> 香芹酚, 糖尿病肾病, 网络药理学, AKT1, PI3K-AKT 信号通路, 细胞凋亡

Abstract:

The mechanism of carvacrol in the treatment of diabetic nephropathy ( DN) by inhibiting endogenous apoptosis pathway was investigated by network pharmacology and in vivo/ in vitro validation experiments. DN mice and renal tubular epithelial cells (HK-2) model were used to investigate the anti-DN effect of carvacrol. The SwissTargetPrediction database was used to predict the potential targets of carvacrol,and the intersection analysis was performed with the DN disease targets screened from the DisGeNET,GeneCards and OMIM databases to obtain the common targets of carvacrol and disease. The STRING platform and Cytoscape software were used to analyze the targets of interaction,so as to screen out the core targets of carvacrol and verify them. KEGG pathway enrichment analysis and GO function annotation were performed on the above intersection targets using the DAVID database. Finally,the prediction results were verified by animal models. Pharmacodynamic studies showed that carvacrol could significantly reduce the levels of fasting blood glucose ( FBG), blood urea nitrogen (BUN),serum creatinine (SCr) and 24-hour urinary albumin (24 h-U-Alb) in DN mice (P < 0. 01,P < 0. 05). Cell experiments showed that carvacrol could increase the viability of HK-2 cells induced by high glucose and inhibit its apoptosis. Network pharmacology studies have shown that the biological processes of carvacrol mainly involve signal transduction,positive regulation of transcription by RNA polymerase II,negative regulation of cytokine production in inflammatory response,Janus kinase-signal transducer and activator of transcription (JAK-STAT) cell surface receptor signaling pathway and apoptosis signaling pathway. It may act on serine/ threonine kinase 1 (AKT1),albumin (ALB),estrogen receptor 1 (ESR1),Janus kinase 2 (JAK2) and progesterone receptor (PGR). Molecular docking and DARTS-Western blot experiments showed that carvacrol had a good binding with AKT1 and could improve the enzymatic stability of AKT1 protein. Western blot experiments showed that carvacrol could inhibit phosphatidylinositol 3-kinase/ protein kinase B (PI3K-AKT) pathway-related proteins. It downregulated the expression of Cleaved Caspase-3,Cleaved Caspase-9 and B-cell lymphoma 2 (BCL-2) -associated X protein (BAX),and up-regulated the expression of BCL-2. In summary,carvacrol has a potential preventive and therapeutic effect on DN,and its molecular mechanism may be related to targeting AKT1 and regulating PI3K-AKT signaling pathway;inhibition of Caspase cascade,mediated endogenous apoptosis pathway is closely related.

Key words:

"> carvacrol, diabetic nephropathy, network pharmacology, AKT1, PI3K-AKT signaling pathway, apoptosis