直接肾素抑制剂阿利吉仑对肾纤维化小鼠上皮-间质转化的调节作用

目的：探讨直接肾素抑制剂阿利吉仑对单侧输尿管结扎引起的小鼠肾纤维化中上皮-间质转化（EMT）的调节作用，阐明阿利吉仑抗肾纤维化的作用机制。方法：24只雄性ICR小鼠分为假手术组、模型组和阿利吉仑干预组，采用单侧输尿管结扎方法制作小鼠肾纤维化模型。HE染色观察小鼠肾间质纤维化的病理组织形态，Masson染色观察小鼠肾间质纤维化中胶原蛋白的表达情况，免疫组织化学染色法观察胞浆中α平滑肌肌动蛋白（α-SMA）的表达，免疫印迹法检测小鼠肾组织中E钙黏蛋白（E-cad）、α-SMA和Ⅰ型胶原（Col Ⅰ）的表达水平。结果：通过单侧输尿管结扎方法成功制备小鼠肾纤维化模型。HE染色和Masson染色，与模型组比较，阿利吉仑干预组小鼠肾纤维化程度明显降低，胶原蛋白表达明显减少。免疫组织化学染色，阿利吉仑干预组小鼠肾小管间质纤维化区域胞浆内α-SMA阳性显色较模型组明显减少。免疫印迹法，模型组小鼠肾组织中α-SMA和Col Ⅰ蛋白表达水平升高，分别是假手术组的1.79倍和1.95倍；E-cad蛋白表达水平降低，为假手术组的37.23%，组间比较差异有统计学意义（P<0.05）；阿利吉仑干预组小鼠肾组织中α-SMA和Col Ⅰ蛋白表达水平降低，E-cad蛋白表达水平升高，分别是模型组的63.72%、65.54%和2.1倍，组间比较差异均有统计学意义（P<0.05）。结论：阿利吉仑能够通过上调E-cad的表达同时下调α-SMA、Col Ⅰ的表达抑制小鼠肾纤维化中EMT，进而发挥抗纤维化的作用。

Regulatory effect of direct renin inhibitor aliskiren on epithelial-mesenchymal transition of renal fibrosis mice

Objective: To explore the regulatory effect of direct renin inhibitor aliskiren on epithelial-mesenchymal transition(EMT) of the renal fibrosis mice induced by unilateral ureteral occlusion,and to clarify its mechanisms of inhibiting renal fibrosis.Methods: Twenty-four ICR mice were divided into sham group,model group, and aliskiren intervention group. The renal fibrosis models were established by unilateral ureteral occlusion.The pathohistomorphology of renal interstitial fibrosis was observed by HE staining.The collagen protein expression in renal interstitial was observed by Masson staining.The α-smooth muscle actin(α-SMA) expression in the cytoplasm was observed by immnocytochemical staining.The protein expression levels of E-cadherin (E-cad), α-SMA,and type Ⅰ collagen(Col Ⅰ) in the kidney tissue of the mice were detected by Western blotting method.Results: The mouse models of renal fibrosis were successfully produced by unilateral ureteral occlusion.The HE staining and Masson staining results showed that the degree of mouse renal interstitial fibrosis in alikiren intervention group was decreased and the expression level of collagen was reduced compared with model group.The immnocytochemical staining results showed that the expression of α-SMA in renal interstitial fibrosis area in alikiren intervention group was obviously reduced compared with model group.The results of Western blotting methods showed that the expression levels of α-SMA and Col Ⅰ in model group were increased to 1.79 folds and 1.95 folds compared with sham group(P<0.05),while the expression level of E-cad was decreased to 37.23%(P<0.05);the expressions levels of α-SMA and Col Ⅰ were decreased to 63.72% and 65.54% compared with model group(P<0.05),while the expression level of E-cad was increased to 2.1 folds(P<0.05).Conclusion: Aliskiren can inhibit the EMT of renal fibrosis produced by unilateral ureteral occlusion via up-regulation of E-cad expression and down-regulation of expressions of α-SMA and Col Ⅰ.