| ObjectiveTo investigate the protective effect of Shenshuaining Tablet on unilateral ureteral obstruction(UUO)rats renal interstitial fibrosis(RIF)and the influence of Wnt/β-catenin signaling pathway,so as to provide the basis for the future clinical trial of Shenshuaining Tablet in the treatment of unilateral ureteral obstruction in RIF.MethodsForty five male Wistar rats aged 6-8 weeks,weighing 180-220 g,were selected for modeling experiment.Forty five healthy rats were randomly divided into sham operation group(S group,n = 15),model group(U group,n = 15)and Shenshuaining tablet treatment group(T group,n = 15),with 15 rats in each group.After successful modeling,group U and group S were given the same volume of normal saline by gavage every day,and group T was given 0.19 g /(kg · d)Shenshuaining tablets by gavage every day from the first day after operation.Five rats in the three groups were killed on the 7th,14 th and 21 st day after operation.Serum creatinine((CREA),blood urea nitrogen(BUN)and microalbuminuria(m Alb)were detected and compared.The severity of RIF in obstructive nephropathy was observed by HE staining.Masson staining was used to observe the changes of renal interstitial collagen volume fraction(CVF)in patients with obstructive nephropathy.The expressions of Wnt and β-catenin in renal interstitium were detected by immunohistochemistry(IHC),and the average optical density(AOD)was calculated for comparison.Results1.The levels of serum crea and BUN in group U were significantly higher than those in group S at 7,14 and 21 days after operation(P < 0.05),followed by group T,and group S was the lowest(P < 0.05),There was no significant change in 14 days,but with the prolongation of obstruction time,there was significant change in urinary protein level in each group at 21 days.The lowest level of proteinuria was found in group S,followed by group T,and the highest in group U.there was significant difference among the three groups(P < 0.05).2.The severity of RIF in U group and T group was higher than that in S group(P < 0.01);Compared with U group,the severity of RIF in T group was decreased at all time points,but still higher than that in S group(P<0.05).3.In group S,the renal interstitium was normal,and there was no change on 7d,14 d and 21d;in group U and T,the blue staining fibrosis mainly appeared in the renal interstitium,and the CVF of renal interstitium in group U and T was higher than that in group S(P < 0.01),and the score increased with time(P <0.05).The score of T group was higher than that of S group(P < 0.05)and lower than that of U group(P < 0.05).4.The positive expression of Wnt1 and β-catenin was brown,and the strong positive expression was brown.The expression of Wnt1 and β-catenin was negative in group S.The positive expression of Wnt1 and β-catenin in group U and group T mainly appeared in cytoplasm and cell membrane,and the expression of Wnt1 and β-catenin was strong positive in epithelial cells and interstitial cells of renal tubules(P < 0.05),The semi quantitative AOD of Wnt1 and β-catenin were calculated by Image J software for comparative analysis.The value of group S was the lowest,and had no significant change at 7 d,14 d and 21 d(P > 0.05).With the extension of time,the AOD gradually increased(P< 0.05).The value of group T was higher than that of group S(P < 0.05),and lower than that of group U(P < 0.05).The difference between the three groups was statistically significant(P < 0.05).Conclusions1.Shenshuaining tablets can improve the degree of RIF in UUO rats and play a protective role in kidney.2.Shenshuaining tablets may play a protective role in renal function by inhibiting Wnt / β-catenin signaling pathway,which may have a certain guiding value for the new treatment direction of RIF in the future. |
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