芪卫颗粒调节KK-Ay小鼠肾组织USF2表达对肾纤维化的干预研究

摘要

Objective To investigate the effects of Qiweikeli on the expression of USF2 and renal fibrosis in KK-Ay mice with spontaneous type 2 diabetic nephropathy. Methods 60 SPF grade 12-week old male KK-Ay mice were randomly di-vided into three groups, model group, Qiweikeli group, valsartan group,normal control was 20 C57BL/6J mice. With Real time PCR and Western blot detection of renal tissue USF2, by its regulation induced fibrosis factor gene and protein expression of TGF-β1 , and regulation downstream fibrosis protein gene and protein expression. 24 weeks of age, blood glucose, glycosylated hemoglobin, renal function ( BUN, SCr ), urinary protein biochemical parameters were detected; renal structural changes with light microscopy and electron microscopy, renal fibrosis with Masson staining were used for analysis. Results (1)Compared with model group, USF2 and TGF-β1 mRNA and protein were significantly higher in renal tissue of the model group,the ex-pression of TGF-β1 ,Col-IV,FN increased than control group,the degree of fibrosis was significantly severe, renal function de-creased obviously( P < 0. 05 ). (2)Compared with the model group, renal function and urinary protein were improved in Qiweikeli group and valsartan group,renal fibrosis was released, USF2 and TGF-β1 gene and fibrosis protein levels were significantly lowei{ P < 0.05 ). (3)Correlation analysis indicate that USF2 was positively correlated to TGF-β1 ( r = 0. 82, P < 0.05 ). Conclu-sion It provided that the Qiweikeli could reduce the expression USF2 gene and protein in the spontaneous type 2 diabetic KK-Ay mice kidney tissue, this may be one of the mechanisms for its treatment of DN by regulating signal transduction.%目的 探讨芪卫颗粒对自发性2型糖尿病KK-Ay小鼠肾组织USF2表达的影响及对DN肾纤维化的干预机制.方法 将SPF级12周龄雄性KK-Ay小鼠60只造模后随机分为模型组、芪卫颗粒组、缬沙坦组,20只C57BL/6J小鼠为正常对照组.24周龄时,检测各组血糖、糖化血红蛋白、肾功能(BUN、SCr)、尿蛋白;用光镜和电镜观察肾组织的结构变化,Masson染色观察肾脏纤维化程度;RT-PCR检测肾组织中USF2、TGF-β1的mRNA表达,Western blot检测USF2蛋白表达,免疫组化染色观察TGF-β1、Col-IV、FN在肾脏中的表达及分布情况.并用图像定量分析系统进行统计分析.结果 (1)与正常对照组比较,模型组肾组织中USF2和TGF-β1的mRNA表达明显升高,免疫组化显示模型组TGF-β1、ColIV、FN在肾小球和肾小管表达均明显增加,纤维化程度明显加重,肾功能减退(P<0.05).(2)与模型组比较,芪卫颗粒组和缬沙坦组肾功能和尿蛋白得到改善,肾脏纤维化减轻,USF2和TGF-β1基因及纤维化蛋白水平表达均显著降低(P<0.05).(3)相关分析表明,USF2与TGF-β1呈正相关(r=0.82,P<0.05).结论 (1)USF2正向调节TGF-β1表达,刺激TGF-β1所调节的下游纤维化蛋白ColIV、FN表达增加,导致细胞外基质积聚.(2)芪卫颗粒下调KK-Ay小鼠肾组织USF2基因和蛋白表达,负向调节TGF-β1表达,可能是其治疗DN肾纤维化的机制之一.