法舒地尔对腹膜透析大鼠腹膜间皮细胞转分化的影响

摘要

To investigate whether Rho-kinase inhibition has a therapeutic role on the peritoneal mesothelium EMT in a rat model of peritoneal dialysis. Methodology; A rat model of peritoneal dialysis was induced by a daily intraperitoneal infusion of 4. 25% Dianeal. Twenty four SD rats were divided into four groups :N Group is normal control group (n =6) ,PD Group is peritoneal dialysis group (n =6) ,Fas 5 mg (n =6) and Fas 10 mg group (n =6) was daily received respectively intraperitoneal injection of 4. 25% Dianeal and 5 mg/kg or 10 mg/kg fasudil. Immunofluorescence, Western blot and RT-PCR were used to detect the expression of TGF-βl ,α-SMA and E-cadherin in each group. The activity of Rho-kinase was determined by western immunoblotting. Results: Rho-kinase was activated in the peritoneum of the PD group, which was inhibited by fasudil in a dose-dependent manner. Compared with normal group, the expressions of TGF-βl and α-SMA in PD group were significantly increased,and the expression of E-cadherin was also markedly decreased. Compared with PD group, the mRNA and protein expressions of TGF-βl and α-SMA were significantly downregulated in fasudil treatment groups in a dose-dependent manner, while the expression of E-cadherin was markedly upregulated in fasudil treatment groups in a dose-dependent manner. Conclusion: Rho-kinase activation play an important role in peritoneal mesothelium EMT in peritoneal dialysis rats, and the inhibition of Rho-kinase by fasudil can significantly inhibit peritoneal mesothelium EMT and improve peritoneal structure and function.%目的:观察Rho激酶(Rho-kinase)抑制剂法舒地尔对腹膜透析大鼠腹膜间皮细胞转分化的影响.方法:通过每日腹腔注射4.25％腹膜透析液建立腹膜透析大鼠模型.大鼠随机分为4组:正常对照组；腹膜透析模型组；法舒地尔5 mg组；法舒地尔10 mg组.4周后杀检大鼠,取脏层腹膜组织,分别用RT-PCR、Western杂交或免疫荧光方法检测转化生长因子β1(TGF-β1)、α平滑肌肌动蛋白(α-SMA)、E钙黏蛋白(E-cadherin)的表达,同时用Western杂交检测Rho-kinase活性. 结果:法舒地尔呈剂量依赖模式显著抑制Rho-kinase活性.与正常组比较,腹膜透析模型组TGF-β1蛋白及mRNA表达显著上调,转分化指标α-SMA蛋白及mRNA表达明显上调(P＜0.01),E-cadherin蛋白及mRNA显著下调(P＜0.01),腹膜超滤量显著降低.法舒地尔干预后,TGF-β1蛋白及mRNA表达呈剂量依赖模式下调,α-SMA蛋白及mRNA表达呈剂量依赖模式下调,E-cadherin蛋白及mRNA表达呈剂量依赖模式上调,腹膜超滤量显著改善. 结论:Rho-kinase激活在腹膜透析大鼠腹膜间皮细胞转分化中起重要作用.法舒地尔可通过抑制TGF-β1信号通路,阻止腹膜间皮细胞转分化,从而改善腹膜透析大鼠模型腹膜结构和功能.