不同浓度锌对成骨细胞MC3T3-E1骨保护素基因表达和细胞增殖的影响

The effect of different concentration of zinc sulfate on the gene expression of osteoprotegerin and cell proliferation of MC3T3-E1 osteoblasts

目的研究不同浓度锌对成骨细胞骨保护素基因表达和细胞增殖的影响。方法取小鼠的成骨细胞系MC3T3-E1培养并随机分为4组,A组加入10μmol/L硫酸锌;B组加入50μmol/L硫酸锌;C组加入200μmol/L硫酸锌;D组作为空白对照。培养48h后提取细胞RNA,RT-PCR分析成骨细胞骨保护素(OPG)mRNA表达,Elisa检测细胞培养上清中骨保护素表达水平。MTT法测定成骨细胞增殖率。结果 A组、B组、C组与D组细胞骨保护素基因表达比值分别为0.461±0.051、1.068±0.123、0.244±0.044、0.548±0.089,A组与D组差异不明显,B组高于D组(P<0.01),C组低于D组(P<0.01)。Elisa结果与RT-PCR相同。细胞增殖率之间差异有显著性意义,B组高于D组(P<0.01),C组低于D组(P<0.01)。结论 50μmol/L的锌促进成骨细胞骨保护素基因表达和细胞增殖,200μmol/L的锌则抑制成骨细胞骨保护素基因表达和细胞增殖。而10μmol/L的锌对成骨细胞骨保护素基因表达和细胞增殖的影响不大。

Objective To explore the effect of different concentration of zinc on the gene expression of osteoprotegerin and cell proliferation of osteoblasts. Methods The mouse osteoblast cell line MC3T3-E1 were cultured and randomly divided into 4 groups. Sulfate zinc was added into Group A （ 10 μ mol/L） , Group B （50 μ mol/L） , and Group C （200 μmol/L）. Group D was treated as control group. RNA was extracted from cells after 48-hour treatment. The mRNA expression of osteoprotegerin was analyzed using RTPCR method. The expression of osteoprotegerin in cell culture medium was measured using Elisa method. The proliferation rate of osteoblasts was measured using MTT method. Results The mRNA expressions of osteoprotegerin in group A, B, C, and D were 0. 461 ± 0. 051, 1. 068 ± 0. 123, 0. 244 ± 0. 044, and 0. 548 ± 0. 089, respectively. The difference between group A and D was not statistically significant. The expression in Group B was higher than that in group D （P 〈 0. 01 ）. The expression in Group C was lower than that of group D （P 〈 0. 01 ）. The Elisa results were identical with those of RT-PCR. The osteoblast proliferation was significantly different. It was higher in Group B than in group D （P 〈 0.01 ） , and was lower in Group C than in group D （P 〈0.01）. Conclusion Zinc of 50μ mol/L promotes the gene expression of osteoprotegerin and proliferation of osteoblasts. Zinc of 200 μmol/L inhibits the gene expression of osteoprotegerin and proliferation of osteoblasts. There is little effect of 10μmol/L zinc on the gene expression of osteoprotegerin and proliferation of osteoblast.