红芪中5种黄酮类成分对大鼠骨髓间充质干细胞和成骨细胞成骨分化的影响

目的研究红芪中毛蕊异黄酮、芒柄花素、芒柄花苷、异甘草素和美迪紫檀素5种黄酮类成分对大鼠骨髓间充质干细胞(rat bone marrow stromal cells,rBMSCs)和颅骨成骨细胞(rat calvarial osteoblasts,ROBs)成骨性分化的影响。方法贴壁分离法培养rBMSCs;酶消解法分离培养ROBs;MTT法检测rBMSCs和ROBs细胞的增殖情况;试剂盒检测rBMSCs和ROBs的碱性磷酸酶(ALP)活性和Ca~(2+)含量;茜素红染色法检测矿化结节形成。结果 5种成分均能不同程度促进rBMSCs和ROBs的增殖,提高ALP活性,增加Ca~(2+)含量,增加钙化结节面积和数量(P0.05)。其中毛蕊异黄酮促进rBMSCs的成骨分化作用最佳,美迪紫檀素促进ROBs的成骨分化作用最佳,毛蕊异黄酮次之。结论这5种黄酮类成分对rBMSCs和ROBs的成骨功能均有一定程度地改善作用,而毛蕊异黄酮对2种细胞表现的成骨活性均较佳,可发展作为良好的骨诱导活性因子。

Effects of promoting osteogenic differentiation of rBMSCs and ROBs using five kinds of flavonoids from Hedysari Radix

Objective To investigate the effects of five kinds of flavonoids (calycosin, formononetin, ononin, isoliquiritigenin, and medicarpin) from Hedysari Radix on promoting osteogenic differentiation of rat bone marrow stromal cells (rBMSCs) and rat calvarial osteoblasts (ROBs). Methods rBMSCs were isolated according to plastic adherence. ROBs were isolated by enzyme digestion method. The proliferation of rBMSCs and ROBs were detected by MTT assay. ALP activity and calcium content of rBMSCs and ROBs cells were detected by alkaline phosphatase kit and calcium kit. Mineralized nodule formation was detected by alizarin red staining. Results The five components could promote proliferation, increase ALP activity, increase calcium content, and increase the area and number of calcified nodules of rBMSCs and ROBs (P <0.05). Among them, calycosin had the best effect on promoting the osteogenic differentiation of rBMSCs, and medicarpin promoted the osteogenic differentiation of ROBs with the best effect, followed by calycosin. Conclusion Five flavonoids promoted the improvement of osteogenic function, while calycosin has better osteogenic activity on rBMSCs and ROBs and can be used as an excellent osteoinductive factor.