1,25（OH）2D3对人肾小球系膜细胞增殖及PCNA表达的影响

摘要

Objective To investigate the effects of 1,25-dihydroxyvitamin D3 [1,25 (OH)2D3] on PCNA expression and cell proliferation in human glomerular mesangial cells. Methods The cultured human mesangial cells, which was subcul⁃tured 3-8 generations, were randomly divided into four groups: normal control group (plus the DMEM medium containing 5%fetal bovine serum), proliferation in the control group (EGF group, plus 10μg/L of EGF), general intervention group [VD group, plus 10-8 mol/L of 1,25(OH)2D3], proliferation in the intervention group [EGF+VD group, plus 10μg/L EGF and 10-8 mol/L 1,25(OH)2D3] for treatment of 48 h. The cell cycle was detected by flow cytometry,and the expression of PCNA was detected by Western blot assay in four groups. Results (1) Compared with normal control group, G1 phase cells were signifi⁃cantly reduced, S, G2/M phase cells were increased and PI index was higher in EGF group. And G1 phase cells were signifi⁃cantly increased, S and G2/M phase cells were significantly decreased, and PI index was lower in VD group. Compared with the EGF group, G1 phase cells were significantly increased in VD group and EGF+VD group, and S, G2/M phase cells de⁃creased, PI index was lower. (2) Compared with normal control group, the expression of PCNA was higher in EGF group, and lower in VD group. Compared with EGF group, the expression of PCNA was lower in VD group and EGF+VD group. Conclu⁃sion 1,25 (OH) 2D3 inhibits the proliferation of human glomerular mesangial cells by arresting cell cycle and inhibiting the expression of PCNA protein.%目的：探讨1,25-二羟基维生素D3[1,25(OH)2D3]对人肾小球系膜细胞中增殖细胞核抗原（PCNA）表达及细胞增殖的影响。方法体外培养人肾小球系膜细胞，取传代培养至3~8代细胞随机分为4组：正常对照组（加含5%胎牛血清DMEM培养基），增殖对照组（EGF组，加10μg/L的EGF），一般干预组[VD组，加10-8 mol/L 1,25(OH)2D3]，增殖干预组[EGF+VD组，加10μg/L EGF及10-8 mol/L1,25(OH)2D3]，均作用48 h。流式细胞术检测各组细胞周期；Western blot检测各组PCNA表达的情况。结果（1）细胞周期。与正常对照组相比，EGF组G1期细胞明显减少，S、G2/M期细胞增多，增殖指数（PI）较高；VD组G1期细胞明显增多，S、G2/M期细胞减少，PI较低；与EGF组相比，VD组和EGF+VD组G1期细胞增多，S、G2/M期细胞减少，PI较低，差异均有统计学意义。（2）系膜细胞中PCNA蛋白的表达。与正常对照组相比，EGF组PCNA的表达较高，VD组PCNA的表达较低；与EGF组相比，VD组和EGF+VD组PCNA的表达较低。结论1,25(OH)2D3通过阻滞细胞周期、抑制PCNA的表达，从而抑制人肾小球系膜细胞的增殖。