他克莫司对人肾小球系膜细胞增殖的影响

Effects of Tacrolimus on proliferation of human mesangial cells

目的探讨他克莫司对体外培养的人肾小球系膜细胞增殖的影响及可能机制。方法采用四甲基偶氮唑盐法(MTT)检测不同浓度的他克莫司处理后系膜细胞的增殖情况;用流式细胞仪检测不同浓度的他克莫司作用48、72h后系膜细胞细胞周期的改变;用流式细胞仪检测不同浓度的他克莫司作用72h后系膜细胞发生凋亡的情况。结果①浓度为1μmol/L的他克莫司对系膜细胞的增殖无明显抑制作用;作用24、48和72h后,浓度为5～20μmol/L的他克莫司能显著抑制系膜细胞的增殖;随着他克莫司浓度的增大,抑制作用更显著,呈一定的剂量—效应关系;随着他克莫司作用时间的延长,抑制作用更显著,呈一定的时间—效应关系。②5μmol/L的他克莫司作用48、72h,G0/G1期的系膜细胞百分比增高,S期的系膜细胞百分比降低,细胞周期阻滞于G0/G1期。③5μmol/L的他克莫司作用72h,对系膜细胞的凋亡无明显影响。结论他克莫司显著抑制体外培养的人肾小球系膜细胞增殖,且呈剂量和时间依赖性;其可能机制是使系膜细胞的细胞周期阻滞于G0/G1期。

Objective To investigate the effects and possible mechanisms of Tacrolimus on proliferation of human mesangial cells.Methods Human mesangial cells（HMCs） were incubated with different concentrations of Tacrolimus.Inhibition of HMC proliferation was analyzed by MTT assay.Flow cytometry was used to analyze the cell cycles of HMC at 48 and 72 hours,and cell apoptosis at 72 hours.Results While 1 μmol/L Tacrolimus did not have any significant inhibition on HMC proliferation,5～20 μmol/L Tacrolimus inhibited the proliferation of HMC in a doseand time-dependent manner after 24,48 and 72 hours.Incubation with 5μmol/L Tacrolimus led to lowered percentage of S stage cells and increased percentage of G0/G1 stage cells after 48 and 72 hours.At 72 hours,5 μmol/L Tacrolimus incubation had no significant effect on the apoptosis rate of HMCs.Conclusion Tacrolimus may inhibit the proliferation of cultured HMC in a dose-and time-dependent manner,probably by blockage of cell cycle at G0/G1 stage.