The Role Of Resveratrol In Renal Fibrosis Caused By Acute Kidney Injury

PART 1.The role of resveratrol in renal ischemia-reperfusion injury ModelObjective:to study the protection effect of resveratrol on the renal ischemia-reperfusion injury in rats model.Methods:By clamping bilateral renal pedicle blood flow of three-weeks-old Sprague-Dawley?SD?rats for 45 minutes to establish model of renal ischemia-reperfusion injury?IRI?in rats.Using resveratrol the natural agonists of Sirt1 continuous intraperitoneal injection model of renal ischemia-reperfusion injury in rats.All male SD rats were randomly divided into 4 groups,control group?Sham?,resveratrol group?Res?,ischemia-reperfusion injury group?IRI?and ischemia-reperfusion injury+resveratrol group?IRI+Res?,each group of 6 8 only.After 14 days,took the tail vein blood for testing the rat serum creatinine?SCr?,urea nitrogen?BUN?and these rats were killed by cervical dislocation method,the anatomy of the rat kidney specimens.Kidney Masson staining to observe each group fibrosis degree of change.Results:Successfully established model of renal ischemia-reperfusion injury in rats.Res group compared with Sham group of SCr and BUN level differences in blood of rats have no statistical significance?P > 0.05?,IRI compared to Sham group SCr and BUN were significantly increased?P < 0.05?,IRI + Res group contrast IRI SCr and BUN level decreased?P < 0.05?.Kidney tissue Masson staining results showed that there was no evident difference between Res group and Sham group,IRI compared to Sham group fibrosis significantly while IRI + Res compared to IRI group reduced renal fibrosis.Conclusion:resveratrol the natural agonists of Sirt1 can effectively relieve renal ischemia-reperfusion injury in rats and renal fibrosis.PART 2.The mechanism of Sirt1 relieve the acute renal injury fibrosisObjective:To explore the protection effect of Sirt1 on acute kidney injury andpossible mechanism of protection.Methods:With different concentrations of H₂O₂ and resveratrol processing rat renal tubular epithelial cell line NRK-52 E after 24 h,MTT method to detect cell activity,and calculate the corresponding cell survival rate.According to the result of MTT experiment,select a suitable concentration of drugs to treat the NRK-52 E cells,cells were observed under inverted microscope morphological after 24 h,flow cytometry instrument to detect the corresponding cell cycle distribution.Western-blot test Sirt1,TGF-?1 and different levels of CTGF protein expression in 24 h after drug treatment.Results:H₂O₂ in 400,800,1200mmol/L concentration decreased NRK-52 E cell survival rate in rats comparing with the control group?P<0.05?,the Res+H₂O₂ group improved the cell survival rate in contrast to H₂O₂ group?P<0.05?.Compared to control group,cells under the stimulus of H₂O₂ becomes slender,sparsely,cells' form and quantity in Res+H₂O₂ group is better than that of H₂O₂ group.Flow cytometry method to detect the cell cycle,the results showed that the ratio of G2/M phase in H₂O₂800mmol/L group reached 31.25%,much higher than 7.62% in the control group?P<0.05?and 10.23% in the H₂O₂800mmol/L+Res10mmol/L group?P<0.05?.The proportion of G2/M phase in H₂O₂800mmol/L+Res10mmol/L group and Res10mmol/L group compared to control group,has no significant difference?P>0.05?.After the treatment of H₂O₂ in 24 h,Sirt1protein expression levels decreased?P<0.05?,TGF-?1,CTGF protein expression quantity increased?P<0.05?.TGF-?1,CTGF protein expression quantity in Res+H₂O₂ group is lower than that in H₂O₂ group?P<0.05?.Sirt1 protein expression levels in Res group increased comparing the control group?P<0.05?.Conclusion:Sirt1 plays a protective role of acute kidney injury fibrosis,its mechanism of action may be through reducing the abnormal increased cells of G2/M phase in damaged renal tubular epithelial.