益肾胶囊对糖尿病肾病大鼠肾小管间质sFRP-1表达的影响

Effect of Yishen Capsule on sFRP-1 in Renal Tubule-interstitium of Rats with Diabetic Nephropathy

目的:观察益肾胶囊对糖尿病肾病大鼠肾小管间质Wnt通路抑制因子-分泌型卷曲相关蛋白-1(sFRP-1)表达的影响。方法:采用一次性腹腔注射链脲佐菌素(STZ)法诱导建立糖尿病肾病(DN)模型,实验分组为:正常对照组(对照组)、DN模型对照组(模型组)、益肾胶囊组、氯沙坦组,每组8只。益肾胶囊组每只大鼠灌胃益肾胶囊625 mg.kg-1.d-1,氯沙坦组每只灌胃氯沙坦20 mg.kg-1.d-1,正常组和模型组每日给予等量生理盐水灌胃,于2、4、8、12周末测定24 h尿蛋白定量,于治疗12周后测定各组血糖、血尿素氮(BUN)、肌酐(Scr),同时观察肾小管病理损伤,并采用免疫组化和实时荧光定量PCR方法观察大鼠肾小管间质中sFRP-1、Wnt通路关键调节因子β-catenin表达情况及肾组织中sFRP-1、β-catenin mRNA的表达。结果:12周末,与对照组相比,模型组大鼠血糖、24 h尿蛋白定量、肾小管损伤指数均显著上升,免疫组化示:肾小管间质中sFRP-1表达增加,β-catenin出现胞浆或(和)核表达,实时荧光定量PCR结果显示二者mRNA表达上调(P<0.05);益肾胶囊治疗后,与模型组相比,肾小管间质中sFRP-1表达进一步增加,肾组织中sFRP-1 mRNA进一步上调,β-catenin胞浆或(和)核表达减少,β-catenin mRNA表达下调(P<0.05)。结论:益肾胶囊可能通过上调sFRP-1在DN大鼠肾小管间质的表达,发挥对DN大鼠肾脏的保护作用。

Objective：To investigate the effect of Yishen Capsule on secreted frizzled related protein-1（sFRP-1）,a Wnt pathway inhibitor,in renal tubule-interstitium of rats with diabetic nephropathy. Methods：The rats models of diabetic nephropathy were established by injecting with streptozotocin intraperitoneally.Wistar femal rats were randomly divided into 4 groups： the control group,the diabetic nephropathy model group,the Yishen Capsule treated group and losartan treated group,each group have 8 rats.The rats in Yishen Capsule treated group were given Yishen Capsule 625 mg/kg through intragastric administration once daily for 12 weeks.The rats in each group were scarificed at the end of 12th week after model established.Blood glucose,blood serum creatinine,blood ureanitrogen,24-h urinary protein were measured,kidneys were processed for Masson-stain as well as immunohistochemistry to observe morphological changes and the expression of sFRP-1,β-catenin.Real-time fluorescent quantitative PCR analyses the expressions of sFRP-1 mRNA,β-catenin mRNA. Results：At the end of 12th week,the expressions of sFRP-1,β-catenin in cytoplasm or nucleus,sFRP-1 mRNA,β-catenin mRNA were significantly increased in the diabetic nephropathy model groups,which was accompanied by higher levels of blood glucose,24-h urinary protein,Scr,Bun and pathology changes compared with the control group（P0.05）,after treatment of Yishen capsule,the expressions of sFRP-1,sFRP-1 mRNA up-regulated further,the expressions of β-catenin in cytoplasm or nucleus,β-catenin mRNA and the 24-h urinary protein,Scr,Bun and pathology changes significantly attenuated compared with the diabetic nephropathy model groups. Conclusion：The mechanism that Yishen capsule prevented fibrosis of renal interstitium may be due to up-regulation the expression of sFRP-1 in renal tubule-interstitium.