盐酸二甲双胍对成骨细胞增殖、BMP-2及Cbfa-1基因表达的影响

Effect of metformin (MF) on the proliferation,bone morphogenetic protein-2 (BMP-2) and core binding factor al (Cbfa-1) gene expression in cultured rat osteoblast

目的初步研究二甲双胍(MF)在体外对小鼠颅盖骨成骨细胞增殖、骨形态发生蛋白-2(BMP-2)及核心结合因子(Cbfa-1)mRNA表达的影响,探讨二甲双胍对骨代谢的可能作用机制。方法 (1)分离培养原代颅盖骨成骨细胞并对其进行鉴定。(2)以乳鼠成骨细胞为体外实验模型,不同浓度(0、50、100、200、400μmol/L)的MF干预体外培养的成骨细胞24h后,MTT法检测成骨细胞的增殖能力,实时荧光定量PCR法检测成骨细胞BMP-2及Cbfa-1基因表达。结果二甲双胍干预成骨细胞24h后,可促进成骨细胞的增殖,在浓度400μmol/L的OD值最大为0.298±0.047(P<0.05);可促进BMP-2及Cbfa-1mRNA的表达,呈剂量效应关系。结论二甲双胍可促进成骨细胞的增殖和分化,可能通过调节BMP-2及Cbfa-1的表达,从而促进骨的形成。

Objective To study the effect of mefformin on proliferation, bone morphogenetic protein-2 （BMP-2） and core binding factor-1 （ebfa-1） gene expression in cultured rat calvarial osteoblasts. And the possible mechanism of action on osteoblats was investigated. Methods （1） Primary ealvarial osteoblasts were isolated, cultured and identified. （2） The rat os teoblasts were cultured in vitro, cultured osteoblasts were treated with metformin in different concentrations （0, 50, 100, 200, 400 μmol/L） after 24 hours. Cell proliferation was assessed by MTI＇ and the expression of BMP-2 and Cbfa-1 mRNA were analyzed by realtimuantitative PCR. Results Osteoblasts were treated with metformin after 24 hours, Metformin could promote osteoblast proliferation, the OD of 400 μmol/L concentrations was the biggest, which was 0.298±0.047 （P 〈 0.05）, and the expression of BMP-2 and Cbfa-1 mRNA were increased, in concentrationdependent manner. Conclusion Met- formin induces the proliferation, differentiation of osteoblast probably by increasing the expression of BMP-2 and Cbfa-1 mRNA.