芪卫颗粒对KK-Ay小鼠肾组织TGF-β_1/Smads信号转导通路的影响

Effect of Qiwei Granules on TGF-β_1/Smads Signaling Pathway in Renal Tissue in KK-Ay Mice

目的探讨芪卫颗粒延缓糖尿病肾病(DN)肾间质纤维化的分子作用机制。方法自发性2型糖尿病KK-Ay小鼠45只,采用高脂饲料喂养制备DN模型。将造模成功小鼠随机分为模型组、中药组[芪卫颗粒20g/(kg·d)灌胃]、西药组[缬沙坦胶囊10mg/(kg·d)灌胃],每组15只,另设C57BL/6J小鼠15只作为正常组,连续给药12周。采用免疫组织化学法检测各组小鼠肾组织中转化生长因子β1(TGF-β1)表达,免疫印迹法检测磷酸化Smad2(P-Smad2)、磷酸化Smad3(P-Smad3)、Smad7及整合素连接激酶(ILK)的表达,实时荧光定量PCR法检测ILKmRNA表达。结果模型组小鼠肾组织中TGF-β1、P-Smad2、P-Smad3、ILK及ILKmRNA表达较正常组明显增加(P<0.01),Smad7表达明显下降(P<0.01)。中药组及西药组TGF-β1、P-Smad2、P-Smad3、ILK及ILKmRNA表达较模型组显著减少(P<0.05或P<0.01),Smad7表达明显增加(P<0.01)。中药组P-Smad3表达较西药组增加(P<0.05),其余指标两组比较差异无统计学意义(P>0.05)。结论芪卫颗粒通过下调TGF-β1、P-Smad2、P-Smad3、ILK的表达,上调Smad7的表达,影响TGF-β1/Smads信号通路,从而延缓了DN肾脏纤维化的发生发展。

Objective To research the molecular mechanism of Qiwei Granules [granules of Huangqi （RadixAstragali） and Weimao （Euonyrni Ramulus ）] to treat interstitial fibrosis in diabetic nephropathy （DN）. Methods KK-Ay mice with spontaneous type 2 diabetes were fed with high-fat fodder to establish DN models. Forty-five successful model mice were randomized into model group, Chinese medicine group and western medicine group, with 15 in each. The Chinese medicine group was gavaged with 20 g/（kg 〈 d） crude drug of Qiwei Granules and the western medicine group was gavaged with 10 mg/ （kg 〈 d） valsartan capsules. Fifteen male C57BL/6J mice were chosen as the normal group. The administration lasted for 12 weeks. The expressions of transforming growth factor-β1 （TGF-β1）, phosphorylated-Smad2 （P-Smad2）, P-Smad3, Smad7, integrin-linked kinase （ILK） and ILK mRNA were examined with immunohistochemistry, Western blot and quantitative real-time polymerase chain reaction respectively. Results Comparing with the normal group, the expressions of TGF-β1, P-Smad2, P-Smad3, ILK and ILK mRNA in the model group were significantly increased （P〈0.01）, but the expression of Smad7 was significantly decreased （P〈0.01）. Comparing with the model group, the expressions of TGF-β1, P-Smad2, P-Smad3, ILK and ILK mRNA in both administration groups were significantly decreased （P〈0. 05 or P〈 0.01）, but the expression of Smad7 was significantly increased （P〈0.01）. Comparing with the western medicine group, the expression of P-Smad3 was significantly increased in the Chinese medicine group （P 〈 0.05）. Conclusion Qiwei Granules can block TGF-β1/Smads signaling pathway and delay the progression of renal interstitial fibrosis by down-regulating the expressions of TGF-β1, P-Smad2, P-Smad3 and ILK and up-regulating the expression, of Smad7.