不同浓度的骨碎补总黄酮对大鼠骨髓间充质干细胞向成骨细胞分化的影响

Effect of rhizoma drynariae total flavonoids on osteogenesis in cultured bone masenchymal stem cells

目的:通过在体外条件下对大鼠骨髓间充质干细胞(BMSCs)实施干预诱导,探寻骨碎补总黄酮对BMSCs的成骨诱导分化能力及不同浓度下可能存在的剂量效应关系。方法:应用全骨髓贴壁培养法对BMSCs进行分离、纯化,并通过流式细胞术对BMSCs进行鉴定,再通过设立含不同浓度骨碎补总黄酮条件培养基,分组对BMSCs进行干预,观察各组7d和14d碱性磷酸酶(ALP)的表达,同时通过ALP染色和矿化结节染色进行定性分析。结果:用全骨髓贴壁法成功分离获得了均一稳定的BMSCs,各浓度骨碎补总黄酮作用BMSCs7d和14d后,诱导不同程度的ALP表达,经10-5g/L骨碎补总黄酮诱导,ALP染色及矿化结节染色均阳性。结论:低浓度骨碎补总黄酮能促进BMSCs向成骨细胞分化,其ALP生成量随着骨碎补总黄酮浓度的降低大致呈增加趋势。

AIM： To explore the effects of total flavonoids of rhizoma drynariae on osteogenic differentiation of bone mesenchymal stem cells （BMSCs） by intervening rat BMSCs with osteogenesis differentiated induction in certain conditions in vitro. METHODS： BMSCs were isolated and purified by the whole marrow culture method, and identified by fow cytometry. BMSCs were treated with different concentrations of total flavonoids of rhizoma drynariae. The expression of alkaline phosphatase （ALP） was quantitatively detected at 7th and 14th day after intervention by ALP and mineralized nodules straining methods. RESULTS： Uniform and stable BMSCs were separated successfully by the whole marrow culture method. After intervention for 7 days and 14 days, the different concentrations of total flavonoids of rhizoma drynariae mani- fested different expression levels of ALP. BMSCs induced by rhizoma drynariae total flavonoids at dose of 10^-5 g/L were all positive with ALP and mineralized nodules straining methods. CONCLUSION： Total flavonoids of rhizoma drynariae at lower concentration promote the osteogenic differentiation of BMSCs. The expression of ALP exhibits a gradually increased tendency accompanied with a decrease in the concentration of total flavonoids of rhizoma drynariae.