Effects Of Tongxinluo On P38MAPK Pathway In Kidney Of Spontaneous T2DM KK-Ay Mice

Objective Investigating the mechanism of Tongxinluo(TXL)to treat fibrosis in diabetic nephropathy by studying the effects of TXL on the expressions of p38 MAPK pathway,,FN and Col-Ⅳ,both in renal tissue of spontaneous T2 DM KK-Ay mice and in glomerular mesangial cells cultured in high glucose.Materials and Methods In vivo,45 spontaneous T2 DM KK-Ay mice were randomly divided into three groups : model group,TXL group and valsartan group,with 15 mice in each group.15 C57BL/6J mice were chosen as the normal group.Blood glucose,renal function(Scr,Urea)as well as urinary micro-albumin biochemical parameters were detected;TGF-β1,p38,P-p38,FN,Col-were detected with Western blot.Ⅳ In vitro,glomerular mesangial cells of SV40 MES 13 mice were used to establish diabetic status by high glucose stimulation.FN and Col-IV were detected by enzyme-linked immunosorbent assay(ELISA)kit in the supernatant of cultured cells.The expression of TGF-beta 1 and p38 MAPK pathway related proteins p38 and P-p38 were detected by WB.Results: 1.Compared with the normal control(NC)group,the fasting blood glucose,body weight,kidney weight and kidney weight index were increased in diabetic mice(P<0.05).Compared with the model group,the body weight,kidney weight,kidney weight index decreased in Tongxinluo group and valsartan(V) group(P<0.05),there was no significant difference between the two groups(P(29)0.05);There was no significant change in fasting blood glucose(P(29)0.05).2.Compared with the NC group,24 h m ALB,SCr,Urea increased significantly in diabetic mice(P<0.05).Compared with the model group,each index level significantly decreased in TXL group and V group(P<0.05),there was no significant difference between the two treatment groups(P(29)0.05).3.Compared with the NC group,the level of FN,Col-Ⅳ,TGF-beta 1 and P-p38 significantly increased in diabetic mice(P<0.05).Compared with the model group,Tongxinluo and valsartan decreased FN,Col-Ⅳ,TGF-beta 1 and P-p38 protein expression(P<0.05)for there was no significant difference between the two treated groups(P(29)0.05).4.Compared with the NC group,FN and Col-protein expression increased in the Ⅳcell supernatant of high glucose groups(P<0.05);compared with the model group,FN,Col-Ⅳexpression decreased in both treatment groups(P<0.05),for there was no significant difference(P(29)0.05).5.Compared with the NC group,the expression of TGF-beta 1 and P-p38 protein was significantly increased in high glucose groups(P<0.05);compared with the model group,P-p38 expression decreased in each treatment group(P<0.05)for no significant difference between the treatment groups(P(29)0.05).Conclusion: 1.Tongxinluo can reduce proteinuria in T2 DM KK-Ay mice,reduce serum creatinine,urea and improve renal function.2.In vivo and vitro,the results showed that the expression of FN and Col-in renal Ⅳtissue of T2 DM KK-Ay mice and mesangial cells could be significantly inhibited by Tongxinluo.3.In vitro and vivo,Tongxinluo can effectively regulate the p38 MAPK pathway,decrease the phosphorylation of p38,reduce the expression of TGF-beta 1 and the accumulation of extracellular matrix components as well as inhibit renal fibrosis.