氟在体外对成骨细胞增殖能力及细胞周期的影响

The Effect of Fluoride on Proliferation and Cell Cycle of Osteoblasts in Vitro

目的 :建立成骨细胞体外培养模型 ,并观察不同浓度的氟对成骨细胞的影响 ,为氟中毒的机制研究提供实验依据。方法 :用幼兔肋骨培养成骨细胞 ,并纯化、鉴定。用不同浓度的氟化钠处理 (2 0 ,16 0 ,2 4 0 ,4 0 0μmol/L)成骨细胞 ,MTT法测定氟对成骨细胞增殖活性的影响 ;流式细胞术测定细胞周期变化及凋亡情况。 结果 :低浓度的氟化钠 (2 0 μmol/L)在体外可显著促进成骨细胞增殖 (P <0 0 1) ,不引起成骨细胞的凋亡 ,可使S期和G2 /M期的细胞明显增多 ;高浓度的氟化钠 (16 0 ,2 4 0 ,4 0 0 μmol/L)则抑制成骨细胞的增殖 (P <0 0 1) ,引起成骨细胞的凋亡 (P <0 0 1和P <0 0 5 ) ,G2 /M期细胞明显减少。结论 :(1)成功建立了氟中毒的体外细胞模型 ;(2 )低浓度的氟化钠可以促进成骨细胞的增殖。高浓度的氟化钠抑制成骨细胞的增殖 ,诱导细胞的凋亡 ,抑制细胞由S期向G2 /M期转化。

Objective: To culture the osteoblasts and study the influence of fluoride on osteoblasts in vitro. Methods: Osteoblasts were cultured, purified and appraised, then dealt with various concentrations of fluoride (20, 160, 240 and 400 μmol/L). The proliferation of osteoblasts was observed with MTT method, and the changes of cell phase and apoptosis were investigated using flow cytometry. Results: Fluoride of low concentration (20 μmol/L) promoted the proliferation of osteoblasts in vitro (P<0.01) and the cells in S and G2/M phase increased markedly. It did not increase the apoptosis of osteoblasts. With the increasing concentrations of fluoride (160,240 and 400 μmol/L), the proliferation of osteoblasts was inhibited. The apoptosis was induced and the cell transformation from S phase to G2/M phase was also inhibited. Conclusions: 1) The model of cell fluorosis has been established in vitro successfully; 2) Fluoride of low concentration could promote the proliferation of osteoblasts and increase the cell number in S and G2/M phases; Fluoride of high concentrations could inhibite the proliferation of osteoblasts and induce the apoptosis; 3) Fluoride has two-phase effects in vitro.