Nephrology Department, Renmin Hospital, Wuhan University, China.
Biomed Pharmacother. 2010 Feb;64(2):124-9. doi: 10.1016/j.biopha.2009.06.001. Epub 2009 Oct 23.
To investigate the influence of fasudil on the epithelial-mesenchymal transdifferentiation of renal tubular epithelial cells from diabetic rats and explore the mechanisms of this effect. Wistar rats were randomly divided into the following three groups: control, diabetes and fasudil-treatment. All rats were sacrificed after three months of feeding with or without fasudil treatment. Pathological changes to the glomeruli and renal interstitium were studied using Periodic acid-Schiff's staining and Masson staining, respectively. Expression of ROCK1, alpha-SMA, E-cadherin and the distribution of beta-catenin in rat renal cortex were revealed by immunohistochemistry. Changes in the MYPT1 phosphorylation profile and alpha-SMA, E-cadherin and membrane beta-catenin expression were revealed by western blot. Changes in the levels of ROCK1, E-cadherin and total beta-catenin mRNA expression were analyzed by real-time PCR. Fasudil treatment notably attenuates renal interstitial fibrosis in diabetic rats. Compared to the control rats, diabetic rats showed elevated phosphorylation of MYPT1, increased expression of ROCK1 and alpha-SMA, decreased expression of E-cadherin and membrane beta-catenin, and increased expression of ROCK1 and total beta-catenin mRNA, decreased expression of E-cadherin mRNA. Fasudil treatment of diabetic rats resulted in attenuated MYPT1 phosphorylation, decreased ROCK1 and alpha-SMA expression, increased E-cadherin and membrane beta-catenin expression, and reduced ROCK1 and total beta-catenin mRNA expression, increased expression of E-cadherin mRNA. In conclusion, fasudil may reduce the epithelial-mesenchymal transdifferentiation and renal interstitial fibrosis in diabetic rats through a mechanism by which ROCK activity is inhibited, which further facilitates the recovery of the cell-cell adhesions among renal tubular epithelial cells and adhesion complex formation.
探讨法舒地尔对糖尿病大鼠肾小管上皮细胞上皮-间质转化的影响，并探讨其作用机制。
Wistar 大鼠随机分为对照组、糖尿病组和法舒地尔治疗组。喂养 3 个月后，分别给予或不给予法舒地尔治疗，然后处死所有大鼠。分别采用过碘酸希夫（Periodic acid-Schiff，PAS）染色和 Masson 染色观察肾小球和肾间质的病理变化，免疫组化法检测肾皮质 ROCK1、α-SMA、E-钙黏蛋白的表达以及β-连环蛋白的分布，Western blot 法检测 MYPT1 磷酸化及α-SMA、E-钙黏蛋白和膜型β-连环蛋白的表达变化，实时荧光定量聚合酶链反应（polymerase chain reaction，PCR）法检测 ROCK1、E-钙黏蛋白和总β-连环蛋白 mRNA 表达的变化。
法舒地尔治疗显著减轻糖尿病大鼠肾间质纤维化。与对照组大鼠相比，糖尿病大鼠 MYPT1 磷酸化水平升高，ROCK1 和α-SMA 表达增加，E-钙黏蛋白和膜型β-连环蛋白表达减少，ROCK1 和总β-连环蛋白 mRNA 表达增加，E-钙黏蛋白 mRNA 表达减少。法舒地尔治疗糖尿病大鼠后，MYPT1 磷酸化减弱，ROCK1 和α-SMA 表达减少，E-钙黏蛋白和膜型β-连环蛋白表达增加，ROCK1 和总β-连环蛋白 mRNA 表达减少，E-钙黏蛋白 mRNA 表达增加。
法舒地尔可能通过抑制 ROCK 活性减少糖尿病大鼠肾小管上皮细胞的上皮-间质转化和肾间质纤维化，进而促进肾小管上皮细胞间细胞黏附的恢复和黏附复合物的形成。
