芪卫颗粒调节USF2及其下游信号分子表达对KK-Ay小鼠糖尿病肾病的干预作用(英文)

Influences of Qi-wei Granules on USF2 and its downstream signal transduction in KK-Ay mice with diabetic nephropathy

目的:探讨自发性2型糖尿病KK-Ay小鼠肾组织TGF-β1、USF2、MMP-9、TIMP-1的表达情况和相互关系,以及芪卫颗粒对糖尿病肾病(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只按血糖值随机分为模型组、芪卫颗粒组、缬沙坦组,20只C57BL/6J小鼠为对照组。24周龄时,行RT-PCR检测肾组织中USF2、TGF-β1的mRNA表达,Western Blot检测TGF-β1、USF2、MMP-9、TIMP-1蛋白表达,免疫组化染色观察TGF-β1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与对照组比较,模型组肾组织TGF-β1、USF2、TIMP-1表达明显升高,MMP-9表达明显降低,免疫组化显示模型组TGF-β1在肾小球和肾小管表达均明显增加(P<0.05)。②与模型组比较,芪卫颗粒组和缬沙坦组TGF-β1、USF2、TIMP-1表达降低,MMP-9表达升高(P<0.05)。结论:①USF2正向调节TGF-β1表达,TGF-β1过表达进而促使MMP-9表达降低、TIMP-1表达升高,MMP-9/TIMP-1比例失衡,细胞外基质(ECM)降解减少而沉积。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,纠正MMP-9/TIMP-1比例失衡,可能是其治疗DN肾纤维化的机制之一。

Objective： To investigate the expression of upstream stimulating factor 2 （USF2）, transforming growth factor- beta 1 （TGF-β1）, MMP-9/TIMP-1 and their relationship with spontaneous type 2 diabetic KK-Ay mice, as well as the mechanism of Qi-wei Granules intervene renal fibrosis. Methods： The KK-Ay mice were SPF grade and 12-week-old, all of them were male. They were randomly divided into three groups including model group, Qi-wei Grannies group, Valsartan group and 20 C57BL/6J mice of control group according to the blood glucose. At 24 weeks of age, USF2 and TGF-βI mRNA expressions were detected by Real-time PCR in kidney tissue, USF2, TGF-β1, MMP-9 and TIMP-1 protein expression were examined by Western blot, the expression and distribution of TGF-β1 were observed by immunohistochemical staining. Results： Compared with the control group, USF2, TGF-β1, tissue inhibitor of metalloproteinase 1 （TIMP-1） were obviously elevated in model group, whereas, MMP-9 expression was significantly reduced, immunohistochemical staining showed that TGF-β1 was expressed in glomerulus and renal tubules and mean optical density （MOD） of TGF-β1 were significantly increased in model group （P〈0.05）. Interestedly, compared with model group, the expression of USF2, TGF-β 1, TIMP-1 were reduced and MMP-9 expression was increased （P〈0.05） in Qi-wei Granules group and Valsartan group. Conclusion： USF2 positively adjusted the TGF-β1 expression. Meantime, the overexpression of TGF-βl not only reduced MMP-9 expression and increased the expression of TIMP-1, but also led to the imbalance of MMP-9/TIMP-1, and the of deposition extracellular matrix （ECM）. Importantly, Qi-wei Granules blocked USF2 expression, negatively regulated TGF-131 and corrected the imbalance of MMP-9/TIMP-1 in KK-Ay mice, which was one of the mechanisms of treating renal fibrosis DN.