王楠兰, 雷露, 敖全, 陈密, 吴彦秋, 张华, 韦艳. 氟化钠对小鼠成骨细胞MC3T3-E1凋亡的影响[J]. 环境与职业医学, 2017, 34(4): 330-336. DOI: 10.13213/j.cnki.jeom.2017.16756
引用本文: 王楠兰, 雷露, 敖全, 陈密, 吴彦秋, 张华, 韦艳. 氟化钠对小鼠成骨细胞MC3T3-E1凋亡的影响[J]. 环境与职业医学, 2017, 34(4): 330-336. DOI: 10.13213/j.cnki.jeom.2017.16756
WANG Nan-lan, LEI Lu, AO Quan, CHEN Mi, WU Yan-qiu, ZHANG Hua, WEI Yan. Effect of sodium fluoride on apoptosis of mice osteoblasts MC3T3-E1 cells[J]. Journal of Environmental and Occupational Medicine, 2017, 34(4): 330-336. DOI: 10.13213/j.cnki.jeom.2017.16756
Citation: WANG Nan-lan, LEI Lu, AO Quan, CHEN Mi, WU Yan-qiu, ZHANG Hua, WEI Yan. Effect of sodium fluoride on apoptosis of mice osteoblasts MC3T3-E1 cells[J]. Journal of Environmental and Occupational Medicine, 2017, 34(4): 330-336. DOI: 10.13213/j.cnki.jeom.2017.16756

氟化钠对小鼠成骨细胞MC3T3-E1凋亡的影响

Effect of sodium fluoride on apoptosis of mice osteoblasts MC3T3-E1 cells

  • 摘要: 目的 探讨高浓度氟化钠(NaF)致小鼠成骨细胞MC3T3-E1细胞凋亡的影响。

    方法 以不同剂量NaF染毒MC3T3-E1细胞,分别培养24和48h,用CCK-8法测定细胞存活率,明确后续实验作用浓度及时间。0、1×103、2×103、3×103μmol/L NaF染毒小鼠成骨细胞MC3T3-E1,培养24、48h后,用流式细胞术检测细胞凋亡;并用实时PCR测定Caspase-3和Caspase-8mRNA的表达及采用免疫印迹法检测Cleaved Caspase-3、Cleaved PARP蛋白的表达。

    结果 CCK-8实验结果显示,随着NaF浓度增加,MC3T3-E1细胞的存活率均随着NaF染毒浓度的升高及染毒时间的延长而下调。1×103 μmol/L时OD值低于对照组,存活率下降,差异有统计学意义(P < 0.05)。1×103μmol/L NaF染毒48h时OD值较24h低,差异有统计学意义(P < 0.05)。与对照组相比,NaF作用24h时,1×103μmol/L NaF染毒组中晚期细胞凋亡率无明显变化,其余各NaF染毒组各期细胞凋亡率均较高,差异有统计学意义(P < 0.05)。随着NaF染毒浓度的升高,MC3T3-E1细胞Caspase-3、Caspase-8 mRNA的表达水平呈上升趋势;且NaF染毒48h后,除1×103μmol/L NaF染毒组的Cleaved PARP蛋白表达外,其余染毒组MC3T3-E1细胞Cleaved Caspase-3、Cleaved PARP的蛋白表达水平上升(P < 0.05)。

    结论 在高浓度NaF诱导细胞凋亡过程中,Caspase介导的信号通路发挥着重要的作用。

     

    Abstract: Objective To test the effects of sodium fluoride (NaF) on apoptosis of osteoblasts MC3T3-E1.

    Methods MC3T3-E1 cells were exposed to different doses of NaF for 24 h and 48 h, respectively. Cell viability was measured by CCK-8 to determine exposure concentrations and time for subsequent experiment. The osteoblasts were treated with 0, 1×103, 2× 103, and 3×103μmol/L NaF for 24 h and 48 h, respectively. Apoptosis was detected by flow cytometry. The expressions of Caspase-3 and Caspase-8 mRNA were detected by real-time PCR and the expression of Cleaved Caspase-3 and Cleaved PARP proteins were detected by Western blot.

    Results The results of CCK-8 assay showed that the survival rate of MC3T3-E1 cells decreased with the increase of NaF concentration and the prolongation of exposure time. At 1×103μmol/L, the OD value of CCK-8 assay was lower than that of the control group, and the survival rate decreased (P < 0.05). The OD value at 1×103μmol/L for 48 h was also lower than that for 24 h (P < 0.05). Compared with the control group, the apoptosis rates at different stages at 24 h of the NaF treatment groups were significantly higher than that of the control group (P < 0.05), except the apoptosis rate at middle and late stages of the 1×103μmol/L NaF group. The expression levels of Caspase-3 and Caspase-8 mRNA showed an increasing trend with the increasing concentration of NaF; and the expression levels of Cleaved Caspase-3 and Cleaved PARP also increased in all NaF groups (P < 0.05), except the expression of Cleaved PARP protein of the 1×103μmol/L NaF group.

    Conclusion The signal transduction mediated by Caspase plays an important role in the apoptosis induced by high concentration of sodium fluoride.

     

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