他克莫司通过稳定细胞骨架和nephrin抑制血管紧张素Ⅱ损伤肾小球足细胞

摘要

Objective To investigate the function and molecular mechanism of tacrolimus in podocyte injury and restoration.Methods Cultured podocytes were stimulated by Angiotensin Ⅱ (Ang Ⅱ) or Ang Ⅱ plus tacrolimus.Cells were collected at different time points (0 h,12 h and 24 h).The distribution of F-actin was observed after immunofluorescence staining,and the protein expression of nephrin and podocin were detected by Western Blot (WB).Results In normal control podocytes,F-actin was arranged in cytoplasm powerfully.Ang Ⅱ induced the disruption and discontinuity of F-actin.Tacrolimus inhibited the effect of Ang Ⅱ,stabilized the regular arrangement the F-actin.Compared to normal cells,the protein expression of nephrin in Ang Ⅱ group significantly decreased at 24 h after stimulation (0.76 ± 0.32 in AngⅡ group vs.1.18 ± 0.40 in normal group,P ＜ 0.05).And tacrolimus stabilized the expression of nephrin protein (1.00 ± 0.19 in treatment group vs.0.76 ± 0.32 in Ang Ⅱ group,P ＜ 0.05).Ang Ⅱ and tacrolimus did not affect the expression of podocin protein.Conclusion Tacrolimus inhibits podocyte injury induced by Ang Ⅱ,stabilizes the regular arrangement of cytoskeleton and protein expression of nephrin.%目的 探讨他克莫司(Tac)在足细胞损伤中的作用机制.方法 以血管紧张素Ⅱ(AngⅡ),或AngⅡ联合Tac干预体外培养足细胞,干预后0、12和24 h收集细胞;行荧光染色观察细胞骨架分布,以WB检测足细胞分子nephrin和podocin蛋白表达.结果 对照组骨架分子F-actin呈强有力丝状排列,AngⅡ引起F-actin的断裂和不连续分布,Tac明显抑制AngⅡ对F-actin的损坏,维持细胞骨架良好的丝状结构.AngⅡ刺激后24h,nephrin表达明显低于对照组,Tac抑制AngⅡ对nephrin的下调(1.00±0.19vs.0.76±0.32,P＜0.05),稳定nephrin正常表达;AngⅡ和Tac干预对podocin表达无明显影响.结论 Tac抑制AngⅡ损伤足细胞,稳定细胞骨架的规律排列和nephrin的正常表达.