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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 study the effects of Genistein, a major component of soy isoflavones, on the proliferation and differentiation of human osteoblasts and the dose-response relationship. METHODS: Human cancellous bone was harvested and cultured. Human osteoblasts were obtained from human osteoblasts by collagenase-trypsin digestion. Purified and subcultured osteoblasts were used as research cells. DMSO control group, estradiol (1 × 10 -8mol / L) positive control group, genistein 1 × 10-9, 1 × 10-8, 1 × 10-7, 1 × 10-6mol / L 4 dose groups. Intervention time were 24,48,72 h. Cell proliferation was detected by MTT colorimetric assay and alkaline phosphatase activity assay was used to detect cell differentiation. Results Human osteoblasts were successfully obtained. Low concentrations of genistein (10-9mol / L) had no significant effect on the proliferation of osteoblasts at various time points, while 10-7mol / L genistein for 72 h significantly stimulated osteoblasts Cell proliferation and differentiation. Spearman correlation analysis found that genistein stimulated the proliferation and differentiation of osteoblasts in a time-and dose-response relationship. Conclusion Genistein can promote the proliferation and differentiation of human osteoblasts, and its promotion effect has the relationship of aging and dose-effect.