1,25-二羟基维生素D_3对人系膜细胞增殖的影响

Effect of 1,25-dihydroxyvitamin D_3 on the proliferation of human glomerular mesangial cells

目的探讨1,25-二羟基维生素D3对人系膜细胞增殖的影响及可能机制。方法体外培养人系膜细胞,分为正常对照组,EGF(10 ng/mL)组,1,25(OH)2D3(10-8mol/L)组,LY294002(2μg/mL)组,雷帕霉素(0.1μg/mL)组,EGF联合1,25(OH)2D3组,EGF联合LY294002组及EGF联合雷帕霉素组,分别采用CCK-8法及流式细胞术检测各组干预48h后对人系膜细胞增殖及细胞周期的影响。结果与对照组比较,EGF组能显著促进人系膜细胞增殖及细胞周期的进程(P<0.01),1,25(OH)2D3组、LY294002、雷帕霉素组、EGF联合LY294002组及EGF联合雷帕霉素组人系膜细胞显著受抑制,细胞周期阻滞于G1期(P<0.01);与EGF组相比,EGF联合1,25(OH)2D3、EGF联合LY294002组及EGF联合雷帕霉素组人系膜细胞均显著受抑制,细胞周期阻滞于G1期(P<0.01);与1,25(OH)2D3组相比,LY294002及雷帕霉素组人系膜细胞均显著受抑制,细胞周期阻滞于G1期(P<0.01)。结论 1,25(OH)2D3可阻滞人系膜细胞细胞周期进程,对人系膜细胞的增殖有显著的抑制作用,且PI3K/Akt/mTOR信号通路可能介导了1,25(OH)2D3对人系膜细胞周期及细胞增殖的抑制作用。

[Objective] To investigate the effect of 1,25-dihydroxyvitamin D3 On the proliferation of cultured human glomerular mesangial ceils and its probable mechanism. [Methods ] The cultured human glomerular mesangial cells were divided into normal control group, EGF （10 ng/mL） group, 1,25-dihydroxyvitamin D3 （10^-8 mol/L） group, LY294002 （2 ug/mL） group, rapamycin （0.1 ug/mL） group, EGF combined 1, 25-dihydroxyvitamin D3 group, EGF combine LY294002 group and EGF combined rapamycin Group. The Cultured human glomerular mesangial cells were treated for 48 hours, cells proliferation was measured by cck-8 assay and the cell cycles were measured by flow Cytometry. [ Results ] Compared with the normal control group, the EGF group can significantly promote the proliferation and cell cycle of human glomerular mesangial cells（P 〈0.01）, but the human glomerular mesangial Cells of 1, 25-dihydroxyvitamin D3 group, LY294002 group, rapamycin group, EGF combined LY294002 group and EGF combined rapamycin group were significantly inhibited and cell cycle were blocked in G1 phase （P 〈0.01）; Compared with the EGF group, the human glomerular mesangial Cells of EGF combined 1, 25-dihydroxyvitamin D3 group, EGF combined LY294002 group and EGF combined rapamycin group were significantly inhibited and cell cycle were blocked in G1 phase （P 〈0.01）; Compared with the 1, 25-dihydroxyvitamin D3 group, the human glomerular mesangial Cells of LY294002 group and rapamycin group were significantly inhibited and cell cycle was blocked in G1 phase （P 〈0.01）. [Conclusion] 1, 25-dihydroxyvitamin D3 can block the human glomerular mesangial cells in cell cycle G1 phase and can inhibit the proliferation of human glomerular mesangial cells and the effect of 1,25-dihydroxyvitamin D3 on the proliferation and cell cycle of cultured human glomemlar mesangial cells may be mediated by the PI3K/Akt/mTOR signaling pathways.