氟对saos-2细胞体外增殖的影响

Effect of fluoride on proliferation of saos-2 cells in vitro

目的探索无血清条件下不同剂量的氟对成骨细胞增殖活性的影响,及以转化生长因子β1(transforming growth factor beta 1,TGF-β1)和骨形成蛋白2(bone morphogenetic protein 2,BMP-2)表达水平间接反映氟对成骨细胞在骨重建过程中的功能的影响。方法以四甲基偶氮唑盐比色法(MTT法)和AlamarBlue法测定氟作用下细胞的增殖。用反转录聚合式酶链反应观察TGF-β1和BMP-2表达水平。结果与对照组相比,氟化钠浓度≤40mg/L时,对细胞均呈现出增殖作用。其中以10mg/L组的促增殖作用最强。当氟化钠浓度>40mg/L时,对细胞以抑制作用为主。两种方法的结果相似。与管家基因β-actin相比较,随氟化钠浓度增高,TGF-β1表达有逐渐增强的趋势,尤其是在80、160mg/L表达最强。BMP-2表达则呈现出在2.5～20mg/L表达明显较对照强,而40、80、160mg/L剂量组的表达较弱。结论氟化钠具有高剂量抑制细胞增殖的作用。即在过量氟作用下正常的成骨细胞合成和分解骨的因子间失去协同一致,偏离正常的动态平衡。

Objective To explore the effect of different doses of fluoride at serum-free conditions on the proliferation of osteoblast activity, and expression of transforming growth factor β1 （TGF-β1） and bone morphogenetic protein 2 （BMP-2）, which indirectly reflect the level of fluoride on osteoblasts in bone in the process of reconstruction of the function. Methods MTT and AlamarBlue methods were used for determination of cell proliferation at different concentration of fluoride. For detecting the expression of TGF-β1 and BMP-2, reverse transcript polymerase chain reaction （RT-PCR） was performed. Results Compared with the control group, proliferation on cell were displayed at sodium fluoride concentration of less than 40 mg/L. Among them, 10 mg/L group played the most powerful role in promoting proliferation. The fluoride concentration over 40 mg/L could inhibit cell growth. The two methods exhibited similar effect. And compared with the housekeeper gene β-actin at different concentration of sodium fluoride, expression of TGF-β1 showed rising trend, particularly in the 80, 160 mg/L. BMP-2 expression shown in the 2.5～20 mg/L expression was stronger, and 40, 80, 160 mg/L dose of weak expression. Conclusions NaF, high-dose inhibition of cell proliferation. That is, the expression of osteoblast synthesis under excess of fluoride, which broken down bone loss and synergy, a departure from normal dynamic balance.