骨碎补总黄酮可促进兔骨髓间充质干细胞的增殖和分化*

背景：骨碎补促进骨损伤愈合疗效确切，但具体作用机制尚不清楚。目的：观察骨碎补总黄酮对兔骨髓间充质干细胞增殖、分化的影响。 方法：冲洗兔股骨和胫骨骨髓腔获取骨髓，采用密度梯度离心法联合贴壁培养法体外纯化扩增兔骨髓间充质干细胞；以四甲基偶氮唑蓝法测定不同浓度骨碎总黄酮对兔骨髓间充质干细胞增殖的影响；经骨碎补总黄酮体外诱导后，行扫描电镜形态学观察，碱性磷酸酶染色、茜素红染色和Von Kossa银染色了解细胞钙化情况。结果与结论：兔骨髓间充质干细胞可以通过密度梯度离心法联合贴壁培养法扩增和纯化；含骨碎补总黄酮血清培养后，四甲基偶氮唑蓝测定结果显示，骨碎补总黄酮浓度为10-6 mmol/L时可明显促进兔骨髓间充质干细胞增殖。经骨碎补总黄酮诱导液诱导，扫描电镜下可见成骨细胞样形态和钙结节形成；细胞碱性磷酸酶染色、钙结节茜素红染色和Von Kossa银染色均呈阳性，提示骨碎补总黄酮可促进兔骨髓间充质干细胞增殖和分化。

Effects of assemble flavone of rhizoma drynariae on proliferation and differentiation of rabbit bone marrow mesenchymal stem cells

BACKGROUND: Rhizoma drynariae is commonly used in orthopedics and showing an accurate effect; however, it specific mechanisms are not very clear.OBJECTIVE: To explore the influences of assemble flavone of rhizoma drynariae on proliferation and differentiation of rabbit bone marrow mesenchymal stem cells (BMSCs).METHODS: The rabbit BMSCs were isolated from femur and tibia bone marrow by combination of gradient centrifugation and different adherent method, then proliferated and purified in vitro. The influence on the rabbit BMSCs proliferation caused by different concentration of assemble flavone of rhizoma drynariae was observed with methyl thiazdyl tetrazolium (MTT) assay; rabbit BMSCs were treated with assemble flavone of rhizoma drynariae inductor in vitro, then to observe the morphology by scanning electron microscope (SEM), identified by alkaline phosphatase staining, and the calcium node formation was detected by alizarin bordeaux staining and von Kossa staining. RESULTS AND CONCLUSION: Rabbit BMSCs could be isolated and cultured by combination of gradient centrifugation and different adherent method. After co-culturing by assemble flavone of rhizoma in vitro, the results of MTT showed that the assemble flavone of rhizoma drynariae with 10-6 mmol/L had an obvious promotion on the rabbit BMSCs proliferation. After assemble flavone of rhizoma drynariae induction, osteoblast-like cell morphology and calcium node formation could be observed under SEM; after dyeing, cells were positive to alkaline phosphatase staining, alizarin bordeaux staining and von Kossa staining. Assemble flavone of rhizoma drynariaecan can accelerate the proliferation and differentiation of rabbit BMSCs.