基于AMPK-SIRT3-BNIP3通路探讨益气养阴活血方对糖尿病肾病大鼠作用机制OA

Objective:To explore the protective mechanism of Yiqi Yangyin Huoxue Recipe on diabetic kidney disease(DKD)rats based on AMPK-SIRT3-BNIP3 signaling pathway.Methods:A total of 42 SPF-grade male SD rats(with body weight ranging from 200~220 g)were used.After 1 week of environmental adaptation,the rats were divided into a blank group(n=6)and a modeling group(n=36).The two groups were fed with normal diet and high-fat/high-sugar diet respectively.A sin-gle intraperitoneal injection of streptozotocin(35 mg/kg)was used to establish the DKD model in the modeling group.The rats that were successfully induced were randomly divided into a valsartan(Diovan)group,a Yiqi Yangyin Huoxue Recipe equal-dose group,a Yiqi Yangyin Huoxue Recipe high-dose group,and a model group.The rats in each treatment group were given the corre-sponding drug doses by gavage,while the blank group and the model group were given the same volume of distilled water.The treatment lasted for 7 weeks.The 24-hour urinary protein(24 h-UTP)and serum biochemical indexes of the rats were detected by biochemical tests,including renal function indexes(SCr and BUN),liver function indexes(ALB and ALT),and glycolipid me-tabolism indexes(GLU,TC,and TG);H&E staining was used to observe the renal pathology;Western blot was used to detect the protein expression levels of AMPK,SIRT3,and BNIP3 in renal tissue;RT-PCR was used to detect the mRNA expression levels of AMPK,SIRT3,and BNIP3 in renal tissue.Results:Compared to the blank group,the levels of ALT,Scr,BUN,TC,TG,GLU,24 h-UTP,and renal tissue BNIP3 protein and mRNA expression levels in the model group rats were signifi-cantly increased(P<0.05),while the levels of ALB and renal tissue SIRT3 and AMPK proteins and mRNA expression levels were significantly decreased(P<0.05),and the renal pathological damage was severe.Compared to the model group,the levels of ALT,Scr,BUN,TC,TG,GLU,24 h-UTP,and renal tissue BNIP3 protein and mRNA expression levels in the treatment groups significantly decreased(P<0.05),while ALB level and renal tissue AMPK and SIRT3 protein and mRNA expression lev-els significantly increased(P<0.05),and the renal pathological damage was significantly alleviated.There was no statistically sig-nificant difference among the treatment groups(P>0.05).Conclusion:The Yiqi Yangyin Huoxue Recipe can effectively reduce 24 h-UTP in DKD rats,protect renal function,and improve renal pathology,thereby delaying the occurrence and development of DKD.The mechanism may be related to the regulation of the AMPK-SIRT3-BNIP3 pathway in renal tissue.