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目的观察金雀异黄酮对缺氧体外培养大鼠骨髓间充质干细胞(BMSCs)增殖与成骨分化的影响。方法采用贴壁筛选法体外培养BMSCs并用三气培养箱建立缺氧模型。设常氧对照组、缺氧对照组和缺氧加药组,其中缺氧加药组细分为1×10-8、1×10-7和1×10-6 mol.L-1组金雀异黄酮组,缺氧处理36 h后分析各组的细胞增殖和成骨性指标包括碱性磷酸酶(ALP)活性、Ⅰ型胶原表达、钙盐沉积量和钙化结节面积等,并用Real Time RT-PCR法检测Runx2和骨形成蛋白(BMP)-2的基因表达情况。结果与缺氧对照组比较,金雀异黄酮抑制BMSCs增殖,显著提高ALP活性、Ⅰ型胶原表达量、钙盐沉积量和钙化结节面积,并提高基因Runx2和BMP-2的表达水平,且呈现出剂量依赖性特点。结论金雀异黄酮具有抑制缺氧BMSCs增殖并促进其成骨分化的作用。
Objective To observe the effects of genistein on the proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) cultured in vitro with hypoxia. Methods BMSCs were cultured in vitro by adherent screening method and hypoxia model was established by three-gas incubator. The normoxia control group, the hypoxia control group and the hypoxia plus drug group were divided into 1 × 10-8, 1 × 10-7 and 1 × 10-6 mol.L-1 groups Sparrow isoflavone group, after 36 h of hypoxia treatment, the cell proliferation and osteogenesis indexes of each group were analyzed including alkaline phosphatase (ALP) activity, type I collagen expression, calcium deposition and calcified nodule area, etc. Real The gene expression of Runx2 and BMP-2 was detected by Time RT-PCR. Results Compared with the hypoxia control group, genistein inhibited the proliferation of BMSCs and significantly increased the activity of ALP, the expression of type I collagen, the deposition of calcium and the area of calcified nodules, and increased the expression of genes Runx2 and BMP-2 Showing a dose-dependent characteristics. Conclusion Genistein can inhibit the proliferation and promote the osteogenic differentiation of hypoxic BMSCs.