Effect Of Tanshinone ?A Combined With DAPT On The Expression Of TRB3,Klotho Protein In Kidney Tissue Of UUO Model Rats

Objective:To observe the effect of tanshinone ?A combined DAPT on the expression of TRB3(tribbles homologous protein 3),Klotho and other proteins in renal tissues of UUO rats,and to deepen the study on the therapeutic mechanism of tanshinone ?combined with DAPT in the prevention and treatment of renal fibrosis,and to provide experimental basis for its clinical application.Methods:Forty male SD rats of SPF grade were randomly divided into 5 groups.The model of renal fibrosis was prepared by left UUO operation,namely,sham operation group(n=8),model group(n=8),tanshinone ?A group(n=8),?-secretase inhibitor(DAPT)group(n=8),tanshinone ?A+DAPT group(compatibility group,n=8).After modeling,the corresponding drugs were given for 14 days.The pathological changes of renal tissue in each group were studied by HE staining.Immunohistochemistry was used to detect the changes of Smad3 and Col III.The protein expression of Smad7,TRB3,Klotho was detected by Western blot(WB).Results:1.HE staining :There was no pathological change in the renal tissue of rats in the sham operation group,and the glomerular and renal tubule structure was normal,and there was no inflammatory cell infiltration.Model group: glomerular structure disorder,tubular atrophy or compensatory expansion,tubulointerstitial widening,interstitial fibrosis obvious,interstitial inflammatory cell infiltration.The renal pathological changes in each administration group were alleviated to varying degrees,and the renal tissue HE scores in each administration group were significantly lower than those in the model group(P<0.05),and tanshinone II A+DAPT group were significantly lower than those in tanshinone II A group and DAPT group(P<0.05).2.Detection of renal tissue by Immunohistochemical: Col ? is expressed in rat renal tissue renal interstitial and tubular epithelial cells;Smad3 in rat renal tissue glomerular and tubular epithelial cells.The average light density of Col ?,Smad3 of renal tissue in model group was significantly higher than in sham group(P<0.05).The average light density of Col ?,Smad3 of renal tissue in tanshinone IIA and DAPT group was lower than in tanshinone II group and DAPT group(P<0.05).3.Detection of renal tissue by WB:Compared with the model group,the expression of TRB3 protein in renal tissue of each administration group decreased significantly(P<0.05),Smad7,Klotho protein expression increased significantly(P<0.05).The expression of TRB3 protein in tanshinone IIA and DAPT group was lower than that in single drug group(P<0.05),Smad7,Klotho protein expression was higher than that of single drug group(P<0.05).Conclusion:Tanshinone IIA and DAPT and its combination can improve fibrosis of renal tissue in UUO rats,and the effect of is better than that of single drug.The mechanism may be related to its decrease UUO protein expression Smad3,TRB3 renal tissue,increase of Smad7,Klotho protein and decrease of Col ? synthesis in rats.