锌协同三羟异黄酮对成骨细胞MC3T3-E1的影响

Effects of Genistein and Zinc on Osteoblastic MC3T3-E1 Cells

目的:观察锌协同三羟异黄酮对成骨细胞MC3T3-E1增殖、细胞中碱性磷酸酶(ALP)含量、骨形成蛋白-2(BMP-2)表达的影响,探讨锌协同三羟异黄酮对骨质疏松的防治作用。方法:采用四甲基偶氮噻唑蓝比色法检测(1×10-7)mol/L、(1×10-6)mol/L、(1×10-5)mol/L、(1×10-4)mol/L的三羟异黄酮以及与(1×10-5)mol/L锌联合作用时对MC3T3-E1增殖的作用;应用Western blot法检测三羟异黄酮与锌联合作用前后,成骨细胞中BMP-2蛋白的表达水平,用比色法检测MC3T3-E1中ALP的含量。结果:锌与三羟异黄酮单独作用或协同作用于MC3T3-E1细胞,其增殖率随着三羟异黄酮浓度的增加和作用时间的延长而升高,(1×10-5)mol/L的三羟异黄酮协同(1×10-5)mol/L的锌作用72 h,其细胞增殖率为(160.1±14.3)%。细胞中的ALP含量及BMP-2的表达也随着三羟异黄酮浓度的增加及作用时间的延长而增加。三羟异黄酮和锌联合作用后,对ALP活性的增强、BMP-2表达的增加作用均较各自单独作用时更为明显(P<0.05)。结论:三羟异黄酮与锌协同作用表现出雌激素效应,可通过促进骨形成蛋白的合成从而促进成骨细胞的增殖、增加骨量。

Objective： To observe the effect of different concentrations of genistein and zinc on cell proliferation and alkaline phosphatase （ALP） activity and bone morphogenetic protein -2 （BMP-2） expression of osteoblast MC3T3-E1 cells and to investigate whether the combination of zinc and genistein has additive effects on the prevention and treatment of osteoporosis. Method： The effect of different concentrations ofgenistein and zinc on proliferation of MC3T3-E1 cells was measured by MTT assay. The expression of BMP-2 was detected by Western blotting assay. Colorimetric method was employed to detect the activity of ALP. Results： The MC3T3-E1 cell proliferation was significantly increased in the presence of zinc and genistein. Genistein promoted MC3T3-E1 cells growth in a time and dose-dependent manner and the rate of cell proliferation was 160.1±14.3 % when the cells treated with 1×10^-5 mol/L genistein and 1× 10^-5 mol/L zinc for 72 h. The ALP content and the expression of BMP-2 in cells treated with genistein were increased in a time and dose-dependent manner. After treatment with Genistein and zinc, the activity levels of ALP and BMP-2 expression were increased more obviously than in the groups treated with genistein or zinc alone. Conclusion： The combination of zinc and genistein displayed the effects of estrogen. By promoting the synthesis of bone morphogenetic protein, it promotes the proliferation of osteoblast cells.