1,25-二羟维生素D_3对人系膜细胞增殖作用的影响

Effect of 1,25 (OH)_2 D_3 on the Proliferation of Human Glomerular Mesangial Cells

目的通过观察1,25-二羟维生素D3〔1,25(OH)2D3〕对Ki-67抗原表达的影响,探讨其对人系膜细胞增殖的抑制作用。方法体外培养人肾小球系膜细胞,随机分为正常对照组、表皮生长因子(EGF)组、1,25(OH)2D3组及EGF联合1,25(OH)2D3组,培养48 h后,应用免疫荧光技术和荧光定量PCR(RT-PCR)检测Ki-67抗原及其mRNA的表达。结果正常对照组、EGF组、1,25(OH)2D3组和EGF联合1,25(OH)2D3组细胞荧光强度分别为(35.958±2.563)、(49.872±2.745)、(22.415±2.277)和(36.567±2.270),4组比较差异有统计学意义(F=123.429,P<0.05)。其中,EGF组荧光强度高于正常对照组,1,25(OH)2D3组荧光强度低于正常对照组(P<0.05);1,25(OH)2D3组和EGF联合1,25(OH)2D3组荧光强度低于EGF组(P<0.05)。以正常对照组为基准,EGF组、1,25(OH)2D3组和EGF联合1,25(OH)2D3组Ki-67 mRNA相对表达量分别为(2.479±0.150)、(0.584±0.031)和(1.176±0.017),4组比较差异有统计学意义(F=51.107,P<0.05)。其中,EGF组Ki-67mRNA相对表达量高于正常对照组,1,25(OH)2D3组Ki-67 mRNA相对表达量低于正常对照组(P<0.05);1,25(OH)2D3组和EGF联合1,25(OH)2D3组Ki-67 mRNA相对表达量低于EGF组(P<0.05)。结论1,25(OH)2D3可抑制人肾小球系膜细胞的增殖,并降低EGF对人肾小球系膜细胞增殖的促进作用。

Objective To investigate the effects of 1,25-dihydroxyvitamin D3〔1,25（ OH）2D3〕on Ki-67 antigen expression in human glomerular mesangial cells and its inhibitive effect on human glomerular mesangial cell proliferation.Methods Glomerular mesangial cells were cultured in vitro and divided into normal control group,epidermal growth factor group（ EGF group）,1,25（ OH）2D3group and combined group of EGF and 1,25（ OH）2D3. After culture for 48 hours, the expression and the mRNA expression of Ki-67 antigen were measured using immunofluorescence technique and RT-PCR.Results The cell fluorescence intensity was（ 35. 958 ± 2. 563）,（ 49. 872 ± 2. 745）,（ 22. 415 ± 2. 277） and（ 36. 567± 2. 270） for normal control group, EGF group,1,25（ OH）2D3group and combined group respectively, with significant difference among them（ F = 123. 429,P 0. 05）. EGF was higher than normal control group in fluorescence intensity（ P 0. 05）,and 1,25（ OH）2D3group was lower than normal control group in fluorescence intensity（ P 0. 05）; 1, 25（ OH）2D3group and combined group were lower than EGF group in fluorescence intensity（ P 0. 05）. With normal control group as standard, the relative expression levels of EGF group,1,25（ OH）2D3group and combined group were（ 2. 479± 0. 150）,（ 0. 584 ± 0. 031） and（ 1. 176 ± 0. 017） respectively,with significant difference among them（ F = 51. 107,P 0. 05）. EGF group was higher than normal control group in the mRNA relative expression level of Ki-67 antigen（ P 0. 05）,1,25（ OH）2D3group was lower than normal control group in the mRNA relative expression level（ P 0. 05）;1,25（ OH）2D3group and combined group were lower than EGF group in the mRNA relative expression level（ P 0. 05）.Conclusion 1,25（ OH）2D3can inhibit the proliferation of human glomerular mesangial cell proliferation and reduce the facilitation of EGF for human glomerular mesangial cell proliferation.