法舒地尔对糖尿病大鼠肾小管上皮细胞转分化的影响

目的 观察法舒地尔对糖尿病大鼠肾小管上皮细胞转分化的影响并探讨其机制。 方法 将实验性Wistar大鼠随机分为正常对照组、糖尿病组、法舒地尔治疗组。3个月后处死动物，PAS染色法观察肾小球病理变化，Masson染色法观察肾间质病理变化；免疫组化观察肾脏皮质ROCKⅠ、α平滑肌肌动蛋白（α-SMA）、E钙黏蛋白（E-cadherin）的变化和β连环蛋白（β-catenin)的细胞定位改变；Western印迹法观察肾脏皮质p-MYPT1、α-SMA、E-cadherin、胞膜β-catenin蛋白的变化；实时定量PCR法观察ROCKⅠ、E-cadherin和总β-catenin的mRNA变化。 结果 法舒地尔治疗可改善肾间质纤维化状况。与正常对照组相比，糖尿病组大鼠肾皮质内p-MYPT1、α-SMA蛋白表达增强（均P < 0.01）；E-cadherin、胞膜β-catenin的蛋白表达减弱（均P < 0.01）；ROCK1、总β-catenin mRNA表达增强（均P < 0.01）；E-cadherin mRNA表达减弱（P < 0.01）。与糖尿病组相比，治疗组p-MYPT1、α-SMA蛋白表达减弱 （均P < 0.01）；E-cadherin、胞膜β-catenin的蛋白表达增强（均P < 0.05）；ROCK1、β-catenin mRNA表达减弱（均P < 0.01）；E-cadherin mRNA表达增强（P < 0.01）。 结论 法舒地尔可能通过抑制Rho激酶活性减轻糖尿病大鼠肾小管上皮细胞转分化和肾间质纤维化，该作用可能与法舒地尔恢复肾小管上皮细胞的黏附特性与紧密连接复合体有关。

Influence of fasudil on the epithelial-mesenchymal transdifferentiation of renal tubular epithelial cells in diabetic rats

Objective To investigate the influence of fasudil on the epithelialmesenchymal transdifferentiation of renal tubular epithelial cells in diabetic rats and to explore the mechanism. Methods Wistar mts were randomly divided into three groups:control,diabetes and fasudil-treatment.All the rots were sacrificed after three months of feeding with or without fasudil.Pathological changes of the glomeruli and renal interstitium were studied by periodic acidSchiff'S staining and Masson staining,respectively.Expression of ROCKI,α-SMA,E-cadherin and the distribution of β-catenin was examined by immunohistochemistry.Changes in the MYPT1 phosphorylation profile and α-SMA,E-cadherin and membrane β-catenin expression were detected bv Western blot.Changes in the levels of ROCKI,E-cadherin and total β-eatenin mRNA expression were analyzed by real-time PCR. Results Fasudil treatment notably attenuated renal interstitial fibrosis in diabetic rats.Compared to the control rats.diabetic rats showed an elevated phosphorylation of MYFF1(P<0.01),increased expression of α-SMA(P<0.01),decreased expression of E-cadherin and membrane β-catenin(P<0.01,respectively)and increased expression of ROCKI,total β-catenin mRNA(P<0.01,respectively),decreased expression of E-cadherin mRNA(P<0.01 ). Fasudil treatment for diabetic rats attenuated MYPT1 phosphorylation (P<0.01), decreased α-SMA expression (P<0.01), increased E-cadherin and membrane (β-catenin expression (P<0.01, respectively), and reduced ROCKI, total β-catenin mRNA expression (P <0.01, respectively), increased expression of E-cadherin mRNA (P<0.01). Conclusions Fasudil may reduce the epithelial-mesenchymal transdifferentiation and renal interstitial fibrosis in diabetic rats through the inhibition of ROCK activity. Such effect further facilitates the recovery of the cell-cell adhesion among renal tubular epithelial cells and the formation of adhesion complex.