摘要
目的:观察黄芪(AM)对糖尿病肾病(DN)大鼠肾间质中Wnt/β-catenin信号通路及转化生长因子-β1(TGF-β1)表达情况的影响,探讨其抗糖尿病肾病肾间质纤维化的作用机制。方法:采用单次空腹腹腔注射60mg/kg链脲佐菌素(STZ)制备糖尿病大鼠模型。4周后测定24h尿蛋白排泄量,以24h尿蛋白定量≥30mg确定DN大鼠模型成功(n=48)。48只DN大鼠随机分为4组:DN模型组(DN组)、黄芪治疗组(DA组)、DN模型组+氯沙坦治疗组(DL组)、DN模型组+黄芪联合氯沙坦治疗组(AL组),每组12只。选取12只健康雄性wistar大鼠作为正常对照组(N组)。连续给药12周后,记录大鼠体重,并检测血清尿素氮(BUN)、肌酐(Scr)及24h尿蛋白定量;处死大鼠,留取肾组织行HE及Masson染色观察肾间质病理改变,采用免疫组织化学技术及实时荧光定量PCR(FQ-PCR)检测肾间质中Wnt4、β-catenin及TGF-β1蛋白及mRNA的表达。结果:与N组比较,12周末DN组大鼠24h尿蛋白定量、血清BUN、Scr水平明显增高,差异有统计学意义(P>0.05)。与N组比较,12周末DN组、DA组、DL组和AL组肾间质Wnt4、β-catenin及TGF-β1蛋白表达量明显增加(P<0.05)。DA组、DL组和AL组作两两比较,AL组24h尿蛋白定量、血清BUN、Scr、Wnt4、β-catenin及TGF-β1下降更为明显,差异有统计学意义(P<0.05)。DA组与DL组相比各指标差异无统计学意义(P>0.05)。FQ-PCR结果显示wnt4、β-catenin和TGF-β1mRNA在各组大鼠肾组织中的表达与Wnt4、β-catenin及TGF-β1蛋白表达趋势一致。结论:黄芪可以降低24h尿蛋白定量,减轻肾间质病理损伤,下调wnt4、β-catenin及TGF-β1在肾间质中的表达,提示黄芪可能通过下调Wnt4、β-catenin及TGF-β1在肾间质中的表达,延缓DN大鼠肾间质纤维化的进程,而发挥肾脏保护作用。
Objective：To investigate the possible mechanism for atragalus mongholicus to protect renal function,the expression of Wnt/β-catenin signaling pathway and TGF-β1 were analysed in STZ-induced of diabetic model.Methods：The diabetes group rats were induced by injection intra-peritoneally with 60 mg/kg body weight STZ in 0.1 mol/L sodium citrate solution（pH 4.3）.The rats in N group were injected with 0.1 mol/L sodium citrate solution.Diabetic model was considered to be successful when 24-hour urinary protein was higher than 30 mg after 4 weeks of the injection（n=48）.After model establishment,these rats were randomly divided into 4 groups：DN group（n=12）,DA group（n=12）,DL group（n=12） and AL group（n=12）.Meanwhile,12 rats were selected to normal group.12 weeks later,body weight,blood glucose,24-hours urine protein quantity and the level of serum creatinine,urea nitrogen were measured,and then all the rats were sacrificed respectively.The methods of immunohistochemistry and real-time fluorescence quantitative PCR were used to detect the expression of Wnt4、β-catenin and TGF-β1 in renal interstitial.Results：24-hour urinary protein,serum creatine and blood urea nitrogen were significantly higher than those of the normal control groups（P〈0.05）.At 12th weeks,in DN group,the expression of Wnt4、β-catenin and TGF-β1 was relatively increased（P〈0.05）.In three treatment group,the expression of Wnt4、β-catenin and TGF-β1 were reduced（P〈0.05）.AL group was lower than the other two,while DA group and DL group have no significantly difference.The tendency of Wnt4、β-catenin and TGF-β1 mRNA expression is tha same with protein expression.Conclusion：AM can decrease 24-hour urinary protein,serum creatine and blood urea nitrogen,alleviating the degree of renal interstitial fibrosis,reducing the expression of Wnt4、β-catenin and TGF-β1,played a role in renal protection.
出处
《中国中西医结合肾病杂志》
2012年第7期571-574,I0001,I0002,共6页
Chinese Journal of Integrated Traditional and Western Nephrology
