染料木黄酮对人成骨细胞增殖及分化影响

Effects of genistein on human osteoblastic proliferation and differentiation

目的研究大豆异黄酮主要成分染料木黄酮(genistein)对人成骨细胞增殖和分化的影响,以及影响的剂量效应关系。方法取材成人松质骨,采用胶原-胰蛋白酶消化法,经纯化和传代培养获得人成骨细胞作为研究细胞来源,设二甲基亚砜(DMSO)溶剂对照组,雌二醇(1×10-8mol/L)阳性对照组,染料木黄酮1×10-9,1×10-8,1×10-7,1×10-6mol/L4个剂量组。干预时间分别为24,48,72 h。用四甲基偶氮噻唑蓝(MTT)比色法检测细胞增殖,碱性磷酸酶活性测定检测细胞的分化。结果成功获得人成骨细胞,低浓度染料木黄酮(10-9mol/L)在各时间点对成骨细胞增殖无明显影响,而10-7mol/L染料木黄酮作用72 h可以明显刺激成骨细胞增殖和分化。Spearman相关分析发现,染料木黄酮刺激成骨细胞增殖分化的作用呈时间和剂量效应关系。结论染料木黄酮能够促进人成骨细胞的增殖和分化,其促进作用存在时效和量效关系。

Objective To investigate the effects of genistein on osteoblastic proliferation and differentiation. Methods Primary osteoblastic cells from human cancellous bone were cultured. Osteoblastic ceils were obtained by trypsin-collagenase digestion. Cells were treated with vehicle （ DMSO ）, genistein （1×10^-9,1×10^-8,1×10^-7,1×10^-6mol/L） and estradiol（1×10^-8mol/L） for 24h,48h,72h,respectively. The cell proliferation and differentiation were investigated with MTT test and the measurement of ALP activity. Results At different time and concentration,gertistein had different effects on osteoblastic cell proliferation. Genistein at 10^-9 mol/L was no effect, but higher concentrations of genistein（ 10^-7 mol/L） stimulated osteoblast proliferation. Spearman correlation analysis showed that the effects of genistein on osteoblastic cell proliferation were time and dose dependent. Genistein could stimulate human osteoblastic cell differentiation in a dose dependent manner. Genistein at 10^-7 mol/L and 10^-6 mol/L increased the ALP activity in a time dependent manner. Conclusion Genistein can stimulate human osteoblastic cell proliferation and differentiation in a dose- and time-dependent manner.