| Objective: Whether Dapagliflozin(Dapa)could improve renal fibrosis in rats with UUO by regulating mitochondrial function and endoplasmic reticulum stress is the aim of this study.To provide experimental basis for clinical prevention and treatment and delay of renal fibrosis.Methods: Eighteen seven-week-old male Sprague Dawley rats weighing about 240-260 g were randomly divided into three groups,including Sham group,UUO group and treatment group(UUO+Dapa),after 1 week of adaptive feeding,with 6 rats in each group.During the experiment,body weight(BW)of each group was measured daily.After 7 days,water intake(WI),24 h urine volume(UV)and body weight of the rats were detected by metabolic cage device.Then the rats were sacrificed and blood samples were collected.Blood creatinine(Scr),blood urea nitrogen(BUN)and hypersensitive C-reactive protein(hs CRP)were detected by biochemical analyzer.After making the kidney tissue sections,then HE staining and Masson staining were used to analyze renal tubule interstitial fibrosis.The morphology of mitochondria and endoplasmic reticulum were analyzed by electron microscopy.Western blots were used to detect the expression of PINK1,Parkin,P62,SDHA,IRE-1α and CHOP.Results:1.Macroscopic observation: Compared to the Sham group,the kidneys in UUO group were enlarged,pale hydronephrosis,and cortical thinning,and the renal pelvis and calycles were dilated,parenchyma and medulla were atrophied significantly,and the boundary was blurred.The changes of UUO+Dapa group were less than those of UUO group.2.Physiological and biochemical measurements: Compared with Sham group,BW decreased,WI increased and UV increased in UUO and UUO+Dapa groups,and the amplitude was approximately similar.Scr,BUN,hs CRP groups had no significant difference.3.HE staining,electron microscopy and Masson staining of renal tissues: Compared with Sham group,renal tubules in UUO group were atrophied,necrotic and vacuolated with thickening of basement membrane,inflammatory cell infiltration and interstitial fibrosis under HE staining.The swelling,lysis and necrosis of the epithelial cells of the obstructed renal tubules and the shedding of microvilli were observed under electron microscope.Masson staining showed vacuolation of renal tubules,collagen deposition and fibrosis formation in UUO group.Compared to UUO group,the results of the above changes were prominently improved in UUO+Dapa group.After Masson staining,the renal tubule interstitial fibrosis score in UUO+Dapa group was significantly reduced by quantitative analysis system.4.Morphological observation of mitochondria and endoplasmic reticulum under electron microscopy: Compared with Sham group,the structure and function of mitochondria in UUO group were damaged,which were expressed as a decrease in the number and size of mitochondria,ridge expansion,vacuolation,mitochondrial fusion,mitochondrial phagocytosis,and pathological division into multiple subcellular organelles.Quantitative analysis showed that Dapa retained the number and size of mitochondria.Compared with Sham group,ribosome degranulation,apoptotic expansion of the endoplasmic reticulum,and peroxisome vacuolation were observed in rough ER in UUO group.5.Western blotting analysis: Compared with Sham group,the expressions of PINK1,Parkin,P62,IRE-1α and CHOP in UUO group were significantly increased,while the expression of SDHA was reduced.Compared to UUO group,the Protein expressions of PINK1,Parkin,P62,IRE-1α and CHOP in kidney tissues of UUO+Dapa group were significantly decreased,while the expressions of SDHA were significantly increased.Conclusion: Dapagliflozin exerts a certain protective effect on kidney fibrosis,and its mechanism of action connected with the regulation of mitochondrial function and endoplasmic reticulum stress by Dapagliflozin. |
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