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

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-β_1的mRNA表达,Westernblot检测USF2蛋白表达,免疫组化染色观察TGF-β_1、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-β_1的mRNA表达明显升高,免疫组化显示模型组TGF-β_1、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-β_1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-β_1呈正相关(r=0.82,P<0.05)。结论 (1)USF2正向调节TGF-β_1表达,刺激TGF-β_1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-β_2表达,可能是其治疗DN肾纤维化的机制之一。

The influences of Qiweikeli on the expression of USF2 and renal fibrosis in KK-Ay mice with diabetic nephropathy

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 divided 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-β_1,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-β_1 mRNA and protein were significantly higher in renal tissue of the model group,the expression of TGF-β_1,Col-Ⅳ,FN increased than control group,the degree of fibrosis was significantly severe,renal function decreased 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-β_1 gene and fibrosis protein levels were significantly lower（P 0.05）.（3）Correlation analysis indicate that USF2 was positively correlated to TGF-β_1（r=0.82,P 0.05）.Conclusion 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.