染料木黄酮对成骨细胞增殖、分化和凋亡影响的实验研究

Effect of genistein on the proliferation, differentiation and apoptosis of the osteoblasts

目的研究染料木黄酮对成骨细胞增殖、分化和凋亡的影响,以阐明其对骨形成的作用机制。方法酶消化和组织块相结合培养获得原代成骨细胞,并以成骨肉瘤细胞株UMR106细胞作对照,加入不同浓度的染料木黄酮后,流式细胞仪和噻唑蓝比色法(MTT)检测成骨细胞细胞周期比例的改变以及凋亡情况;生化法检测细胞内碱性磷酸酶含量的改变。结果流式细胞分析和MTT观测结果表明:染料木黄酮促进了原代成骨细胞由G0(G1)期向S期、G2期和M期的移行过渡,从而促进了原代成骨细胞的增殖。染料木黄酮对成骨肉瘤细胞株UMR106细胞周期无影响,但可诱导UMR106细胞凋亡。碱性磷酸酶检测结果表明:染料木黄酮可促进原代成骨细胞的分化。结论染料木黄酮可在一定程度上促进成骨细胞的增殖和分化,诱导成骨肉瘤细胞凋亡。

Objective To study the effect of genistein on the proliferation, differentiation and apoptosis of the osteoblasts in vitro. Methods The primary osteoblasts (OBs) were obtained from the rat calvaria and the cell line of osteosarcoma -UMR-106 served as control. The cells in the experiment group were grown in 10% (volume fraction) fetal calf serum ( FCS ) + αMEM + various concentrations of genistein. The control groups were grown in 10% FCS +αMEM. The growth of OBs was assessed by flow cytometry and MTT method. The differentiation of OBs was examined by the alkaline phosphatase(ALP) activity. Results Flow cytometry analysis and MTT showed that genistein could prompt primary OB from stage G_0(G_1) to stage S, G_2 or M. By contrast, genistein had no effect on the cell cycle of UMR-106, but could induce its apoptosis. Additionally, the results of ALP activity showed that genistein stimulated the differentiation of primary OB. Conclusions Genistein can stimulate the proliferation and differentiation of the primary osteoblasts in some degree, and induce the apoptosis of osteosarcoma cells.