钙对人系膜细胞增殖及1,25-D3-MARRS蛋白表达的影响

Effects of calcium on proliferation of human mesangial cells and the expression of 1,25-D3-MARRS

目的探讨钙离子对人肾小球系膜细胞(HMC)增殖的影响及维生素D膜相关快速反应结合蛋白(1,25-D3-MARRS)表达的改变。方法①体外培养HMC,给予不同浓度钙离子培养液干预细胞48 h,分为3组:正常对照组(0 mol/L CaCl 2)、低钙组(10^-4 mol/L CaCl 2)、高钙组(1.8×10^-3 mol/L CaCl 2);② CCK-8法检测细胞增殖能力;③免疫荧光法和Western blot法检测各组1,25-D 3-MARRS蛋白的表达;④ RT-PCR法检测各组1,25-D 3-MARRS mRNA的含量。结果①与正常对照组比较,低钙组(10^-4 mol/L) HMC抑制率增高,高钙组(1.8×10^-3 mol/L ) HMC的抑制率降低( P <0.05);② 1,25-D 3-MARRS在各组HMC均有表达,且主要表达在细胞膜,与正常对照组比较,低钙组1,25-D 3-MARRS蛋白表达升高( P <0.05);高钙组蛋白表达降低,但差异无统计学意义;③与正常对照组比较,低钙组 1,25-D 3-MARRS mRNA 表达升高( P <0.05),高钙组mRNA表达降低,但差异无统计学意义。结论低浓度钙离子可以抑制HMC的增殖,其作用机制可能与1,25-D 3-MARRS蛋白的表达增高有关。

Objective To investigate the effects of calcium on the proliferation of human mesangial cells(HMC) and the expression of membrane associated, rapid response steroid binding receptor(1,25-D3-MARRS) in vitro. Methods HMC was cultured in vitro, and cells were treated with different concentrations of calcium ion culture medium for 48 hours. They were divided into three groups: normal control group(group N, 0 mmol/L CaCl2), low concentration calcium ion group(group L, 10-4 mol/L CaCl2), high concentration calcium ion group(group H, 1.8×10-3 mol/L CaCl2). The cell proliferation ability was detected by CCK-8. The expression of 1,25-D3-MARRS protein was detected by immunofluorescence and Western blot. Meanwhile,the mRNA expression of 1,25-D3-MARRS was detected by RT-PCR. Results Compared with group N, the inhibition rate of HMC in group L was increased, and decreased in group H(P<0.05). 1,25-D3-MARRS was expressed in all groups, mainly expressed in cell membrane. The expression of 1,25-D3-MARRS protein and mRNA in group L was higher than that in group N and group H(P<0.05), the expression in group H was lower than that in group N and group L. Conclusion Low concentration of calcium ions may inhibit the proliferation of HMC, by up-regulating the expression of 1,25-D3-MARRS.