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目的探讨活性维生素D3(1,25(OH)2D3)对大鼠系膜细胞增殖的影响。方法体外培养大鼠系膜细胞,随机分为正常对照组、表皮生长因子(EGF)(10 ng/m L)组、1,25(OH)2D3(10-8mol/L)组、EGF联合1,25(OH)2D3组,采用MTT法检测各组干预48 h后对大鼠系膜细胞增殖的影响,流式细胞术检测各组干预48 h后大鼠系膜细胞周期分布情况,免疫荧光法检测各组干预48 h后,大鼠系膜细胞中增生性细胞核抗原(PCNA)的表达情况。结果与正常对照组比较,EGF组能明显促进大鼠系膜细胞增殖,G0/G1期细胞减少,S期及G2/M期细胞增加,且PCNA表达增加,1,25(OH)2D3组大鼠系膜细胞增殖受抑制,G0/G1期细胞增加,S期及G2/M期细胞减少,且PCNA表达降低;与EGF组比较,EGF联合1,25(OH)2D3干预组系膜细胞增殖受抑制,G0/G1期细胞增多,S期及G2/M期细胞减少,且PCNA表达降低。结论 1,25(OH)2D3可阻滞大鼠系膜细胞的细胞周期并抑制大鼠系膜细胞的增殖和EGF对大鼠系膜细胞的促增殖作用。
Objective To investigate the effect of active vitamin D3 (1,25 (OH) 2D3) on the proliferation of rat mesangial cells. Methods Rat mesangial cells were cultured in vitro and randomly divided into normal control group, epidermal growth factor (EGF) 10 ng / m L group, 1,25 (OH) 2D3 10-8 mol / L group, , 25 (OH) 2D3 group. MTT assay was used to detect the effect of each group on mesangial cell proliferation 48 h after intervention. Flow cytometry was used to detect the cell cycle distribution of rat mesangial cells after 48 h intervention. Immunofluorescence Method was used to detect the expression of proliferating cell nuclear antigen (PCNA) in rat mesangial cells 48 h after intervention. Results Compared with the normal control group, the EGF group could significantly promote the proliferation of rat mesangial cells, the number of cells in G0 / G1 phase, the cells in S phase and G2 / M phase increased, and the expression of PCNA increased. In the 1,25 (OH) 2D3 group, The proliferation of rat mesangial cells was inhibited, the cells in G0 / G1 phase increased, the cells in S phase and G2 / M phase decreased, and the expression of PCNA decreased. Compared with EGF group, the proliferation of mesangial cells was significantly inhibited by EGF combined with 1,25 (OH) 2D3 Inhibited, G0 / G1 phase cells increased, S phase and G2 / M phase cells decreased, and PCNA expression decreased. Conclusion 1,25 (OH) 2D3 can block the cell cycle of rat mesangial cells and inhibit the proliferation of rat mesangial cells and the effect of EGF on the proliferation of rat mesangial cells.