摘要
目的探讨环孢素A(CsA)对人系膜细胞增殖的抑制作用及细胞周期蛋白D1、E、p27kip1表达影响。方法人系膜细胞常规体外培养,分为实验组和对照组,实验组加入不同浓度的CsA(0.1,1,5μmol/L)干预。药物作用24,48,72 h后MTT比色法测定细胞增殖情况。药物作用48 h后采用流式细胞仪检测细胞周期。药物作用48 h后Western blot法检测细胞中cyc-lin D1、cyclin E、p27kip1蛋白质表达情况。结果 CsA对人系膜细胞具有时间、浓度依赖性抑制作用;且浓度依赖性阻滞细胞周期,使G0/G1期细胞增多,S期细胞减少(P<0.05)。CsA浓度依赖性下调cyclin D1、cyclin E蛋白,上调p27kip1蛋白的表达(P<0.05)。结论 CsA通过下调cyclin D1、cyclin E蛋白,上调p27kip1蛋白的表达,调节细胞周期调控通路,阻滞细胞于G0/G1期,有效抑制人系膜细胞的增殖。
Objective To explore the effect of CsA on proliferation,apoptosis and cell cycle of human mesangial cells. Methods Human mesangial cells were incubated in vitro with different concentrations of CsA(0,0.1,1,5 μmol/L).The proliferation of human mesangial cells was detected after incubated with different concentrations of CsA for 24,48,72 h by MTT assay,respectively.Cell cycle was analyzed at 48 h after incubation by flow cytometry.The expression of cyclin D1,cyclin E and p27kip1 proteins in cell nucleus was assessed by Western blot. Results The proliferation of human mesangial cells was inhibited in a time-and dose-dependent manner by CsA.The cell cycle was arrested in G0/G1 phase.The percentage of G0/G1 phase cells gradually increased with the rising of CsA concentrations(P0.05).The expression of cyclin D1,cyclin E proteins decreased in cell nucleus with the rising concentrations of CsA,while expression of p27kip1 protein increased significantly(P0.05). Conclusion CsA could inhibit the mesangial cell proliferation by decreasing the expression of cyclin D1,cyclin E proteins and increasing the expression of p27kip1 protein.
出处
《山西医科大学学报》
CAS
2012年第4期245-248,共4页
Journal of Shanxi Medical University