氟铝单独及联合染毒对MC3T3-E1细胞增殖能力及细胞周期的影响

Effects of Fluoride and Aluminum Alone and in Combination Exposure on Proliferation and Cell Cycle of MC3T3-E1 Cells

目的观察不同浓度的氟、铝对体外培养的小鼠颅顶前成骨细胞亚克隆14(MC3T3-Esubclone 14)增殖及细胞周期的影响,以进一步阐明地方性氟中毒机制提供实验依据。方法以10-9～10-3mol/LNaF染毒MC3T3-E1细胞,同时以50μmol/LNaF及5μmol/LAlCl3单独或者联合染毒MC3T3-E1细胞,培养72h。应用CCK-8(cell counting kit-8)观察MC3T3-E1细胞增殖能力的影响;采用流式细胞术检测MC3T3-E1细胞周期变化情况。结果氟无显著促进MC3T3-E1细胞增殖的作用,较高浓度的氟(1mmol/L)抑制MC3T3-E1细胞增殖(P<0.01)。氟铝联合染毒显著刺激MC3T3-E1细胞增殖(P<0.01);G2/M期细胞明显增多,增殖指数(PI)升高(P<0.05),DNA相对含量增高(P<0.05)。结论氟对MC3T3-E1细胞无显著促增殖作用,氟铝联合能显著提高MC3T3-E1的增殖能力,使G2/M期细胞明显增多,促进成骨细胞的增殖分裂,细胞处于活跃生长状态。

Objective To explore the effects of different concentrations of fluoride, aluminum alone and in combination exposure on mice parietal bone cell subclone 14 （MC3T3-E subclone 14）, and to elucidate the pathogenesis of endemic fluorosis. Methods The proliferation of MC3T3-E1 cells exposed to 10^-9-10^-3 mol/L NaF alone, 50 μmol/L NaF and 5 μmol/L AlCl3 alone and in combination ,was measured by CCK-8, and the change of cell cycle was measured by flow cytometry after treatment with various concentrations of fluoride and aluminum. Results Fluoride alone did not promote osteohlast MC3T3-E 1 cells proliferation, higher concentration fluoride inhibited MC3T3-E 1 cells proliferation. Fluoride and aluminum combined exposure （50 μmol/L NaF ＋5 μmol/L AlCl3） stimulated proliferation of MC3T3-E1 cells （P〈0.01）,and significantly induced increase of G2/M phase, PI （proliferation index） and DNA relative content. Conclusion Fluoride does not promote the MC3T3-eells proliferation, aluminium plus fluoride may increase MC3T3-E 1 cells proliferation and affect the cell cycle, which can significantly increase the number of G2/M phase cells.