结缔组织生长因子-siRNA对糖尿病肾病大鼠肾组织中转化生长因子-β1及层黏连蛋白表达的影响

摘要

目的:探讨结缔组织生长因子(CTGF) si-RNA对糖尿病大鼠肾组织中转化生长因子β(TGF-β1)及层黏连蛋白表达的抑制作用.方法:取健康雄性Wistar大鼠,随机分为正常对照组、糖尿病模型组、CTGF-siRNA隐性对照组、CTGF-siRNA实验组.腹腔注射1％链脲佐菌素(STZ)建模.特殊转染载体通过尾静脉注射CTGF-siRNA.免疫组织化学观察肾组织中TGF-β1的表达,RT-PCR方法观察肾皮质中TGF-β1、层黏连蛋白mRNA表达的变化.结果:CTGF-siRNA实验组肾组织中TGF-β1的表达减少,同时肾皮质中TGF-β1、层黏连蛋白mRNA表达也减少.CTGF-siRNA注射结束后第4周,尿蛋白明显降低.结论:CTGF-siRNA对糖尿病大鼠肾组织中TGF-β1、层黏连蛋白表达有抑制作用,缓解肾组织的纤维化作用,同时对糖尿病肾功能有明显改善作用.%Objective:To investigate the effect of CTGF-siRNA on the expression of TGF-J31 and fibronectin in the renal tissue of rat with diabetes. Methods: Diabetes was induced in male Wistar rats by intraperitoneal injection of streptozotocin for consecutive three days. After the development of diabetes, the rats were treated with CTGF-siRNA for every two days for three times. Four weeks after the last dose of CTGF-siRNA,the rats were killed and the kidneys were removed. Immuno-histochemistry was done to observe the protein expression of TGF-β1 in the renal tissues and RT-PCR analysis was also done to examine the mRNA expression of TGF-β1 and fibronectin in the renal cortex. In order to evaluate the renal function, 24 h urine protein was analyzed at day 7, 14, 21 and 28. Results:CTGF-siRNA significantly decreased the expression of TGF-β1 protein and mRNA in the renal tissues and also greatly reduced the fibronectin mRNA expression in the renal cortex. Additionally, there was also significant reduction of urine protein at the 28th day. Conclusion:CTGF-SiRNA can inhibit the increased expression of TGF-β1 and fibronectin in the renal tissues, and also improve renal function in diabetic nephropathy rats.