蜂胶黄酮对大鼠颅骨成骨细胞增殖分化及BMP2表达的影响

Effects of propolis flavonoid on proliferation,differentiation and expression of BMP2 in cultured calvarial osteoblasts from newborn rats

目的探讨蜂胶黄酮对体外培养大鼠颅骨成骨细胞增殖分化以及骨形态发生蛋白2(BMP2)表达的影响。方法提取新生大鼠颅骨成骨细胞制作体外培养模型,用不同浓度的蜂胶黄酮刺激分离培养的大鼠成骨细胞,然后用噻唑蓝(MTT)法测定蜂胶黄酮对体外培养成骨细胞的增殖作用,用氨基安替吡啉测酚法测成骨细胞内碱性磷酸酶(ALP)活性,并通过免疫组织化学方法观察成骨细胞BMP2的表达。结果添加蜂胶黄酮体外培养24h后,100μg/L剂量组较对照组有明显的提高;48h后,5、10和100μg/L剂量组与对照组比,增殖率均显著提高;添加蜂胶黄酮的中剂量组和高剂量组的BMP2表达在24h和48h都显著高于零剂量组。培养72h后,10、50和100μg/L剂量组与对照组比,增殖率均有明显的提高;50μg/L和100μg/L剂量组与对照组相比ALP活性均显著性提高。结论蜂胶黄酮可以增加成骨细胞的增殖和分化,促进骨组织的形成,可用来预防骨质疏松症。

[Objectives]To investigate the effects of propolis flavonoid （PF） on proliferation, differentiation and expression of BMP2 in cultured calvarial osteoblasts from newborn rats. [ Methods ] Osteoblastic cells from the calvari of newborn rat were cultured in vitro. We added propohs flavonoid in suitable concentration into the culture fluid. The proliferation of osteoblastic cells was measured by MTT and activity of ALP was observed with Golden＇s method. The immunohistochemistry assay was used to measure the expression of BMP2. [ Results ] Compared with the control group, the proliferation rate of osteoblastic cells were significantly elevated in group with dose of 100 μg/L PF after 24 hours; in groups with dose of 5, 10 and 100μg/L PF after 48 hours. Compared with the zero-dose group, the expression of BMP2 significantly increased in both middle-dose group and high-dose group after 24 hours and 48 hours. After 72 hours culture, the proliferation rate was significantly improved in groups with dose of 10, 50 and 100 μg/L PF comparing with the control group. The levels of ALP activity were significantly increased in groups with of 50μg/L and 100 μg/L PF. [ Conclusion] The propolis flavonoid may increase the proliferation, differentiation of os- teoblastie cells and stimulates the formation of bone tissue in vitro. It can be used to prevent fl＇om the osteoporosis.