The Expression And Significance Of Sirt6 In A Novel Alcoholic Renal Injury Mouse Model

Objectives:1.To construct and verify a novel alcoholic renal injury mouse model that simulates common human drinking patterns.2.To observe the expression of sirtuin protein family in alcoholic renal injury model,to focus on the expression of SIRT6 and the possibility of sirt6-p53 axis participating in alcoholic kidney injury.Methods:1.Establishment and verification of the new alcoholic kidney injury model:20 healthy male 57BL/6 mice were selected,and after adaptive feeding for 5 days,they were randomly divided into the control group and the alcohol group.The control group was fed with control liquid diet for 10 days,and then acute gavage of 45%dextrin.The alcohol group was fed with eqlal-calorie 4.2%ethanol diet for 10 days,and then acute gavage of 31.5%alcohol.The renal gross structure and renal index of the two groups were observed;The histopathological changes of kidney were observed by H&E staining and PAS staining;The expression of fibrosis and in:flammatory cytokines in renal tissues was detected by Real Time PCR.To determine whether the new alcoholic renal injury model was successfully constructed.2.To explore the sirtuin family located in the nucleus and mitochondria,especially the expression and role of SIRT6 in alcoholic renal injury:Real Time PCR detected the mRNA expression of SIRTs and p53;Immlnofluo'rescence staining and Western Bolt were used to detect the expression position of SIRT6 in kidney tissues and the changes of SIRT6 protein expression level after alcoholic kidney injuryResults:1.Related results of the construction of the novel alcoholic renal injury model:1)the mice in both groups were in good condition,but the renal quality and renal index in the alcohol group were higher than those in the control group(P<0.01).2)H&E and PAS staining in the control group showed normal renal structure.While in alcohol group,the H/E staining showed Proliferation of glomerular mesangial cells,widening of the space between the swelling of renal tubular epithelial cells,The small blood vessels in the interstitium are characterized by hyperplasia,dilatation and edema,surrounded by inflammatory cell infiltration.PAS staining showed glomerular enlargement,mesangial cell and mesangial matrix hyperplasia in the alcohol group.3)compared with the control group,mRNA expressions of TGF-?1 and inflammatory cytokines in the alcohol group were up-regulated(P<0.05).2.Effects of alcohol on sirtuin family and p53 in kidney:1)compared with the control group,the mRNA expressions of SIRT1,SIRT6 and p53 were up-regulated(P<0.05),and the mRNA expressions of SIRT3 and SIRT5 were down-regulated in alcohol group kidney(P<0.05).2)the kidney SIRT6 protein expression in the alcohol group was increased(P<0.001),and the protein expression of p53 was decreased(P<0.01).3)Immunofluorescence staining showed that SIRT6 expression in the alcohol group was more than that in the control group,and SIRT6 expression in renal tubules and mesenchymal cells was more than that in glomeruli.Conclusions:1.By chronic alcohol feeding combined with acute gavage of alcohol successfully established a novel alcoholic kidney injury mouse model.Compared with previous models,this model simulates the common drinking patterns of human beings,and the modeling time is short and repeatable,the mice have a high acceptance degree to ethanol feed,and the mice are in good condition.2.SIRT6 was up-regulated in alcoholic kidney injury,and sirt6-p53 axis may be involved in the inflammatory and fibrotic responses of alcoholic renal injury.