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

作　　者：王楠兰 雷露[1] 敖全 陈密[1] 吴彦秋[1] 张华[1] 韦艳[1] 

机构地区：[1]贵州医科大学公共卫生学院环境卫生学教研室,环境污染与疾病监控省部共建教育部重点实验室,贵州贵阳550004

出　　处：《环境与职业医学》2017年第4期330-336,共7页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金(编号:81560515);贵州省科技厅社会发展攻关计划项目(编号:黔科合SY字[2013]3062号);科技部国家科技支撑计划课题(编号:2013BAI05B03)

摘　　要：[目的]探讨高浓度氟化钠(Na F)致小鼠成骨细胞MC3T3-E1细胞凋亡的影响。[方法]以不同剂量Na F染毒MC3T3-E1细胞,分别培养24和48 h,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、1×10~3、2×10~3、3×10~3μmol/L Na F染毒小鼠成骨细胞MC3T3-E1,培养24、48 h后,用流式细胞术检测细胞凋亡;并用实时PCR测定Caspase-3和Caspase-8 m RNA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Na F浓度增加,MC3T3-E1细胞的存活率均随着Na F染毒浓度的升高及染毒时间的延长而下调。1×10~3μmol/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)。1×10~3μmol/L Na F染毒48 h时OD值较24 h低,差异有统计学意义(P<0.05)。与对照组相比,Na F作用24 h时,1×10~3μmol/L Na F染毒组中晚期细胞凋亡率无明显变化,其余各Na F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Na F染毒浓度的升高,MC3T3-E1细胞Caspase-3、Caspase-8 m RNA的表达水平呈上升趋势;且Na F染毒48 h后,除1×10~3μmol/L Na F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Na F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。[Objective] To test the effects of sodium fluoride （NaF） on apoptosis of osteoblasts MC3T3-E 1. [ Methods ] MC3T3-E1 cells were exposed to different doses of NaF for 24h and 48h, respectively. Cell viability was measured by CCK-8 to determine exposure concentrations and time for subsequent experiment. The osteoblasts were treated with 0, 1 × 10^3, 2 × 10^3, and 3 × 10^3 μmol/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. [ Rostults ] The results of CCK-8 assay showed that the survival rate of MC3T3-E1 ceils decreased with the increase of NaF concentration and the prolongation of exposure time. At 1 × 10^3/amol/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 × 10^3 μmol/L for 48h was also lower than that for 24h （P〈 0.05）. Compared with the control group, the apoptosis rates at different stages at 24h 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 × 10^3 μmol/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）, exeept the expression of Cleaved PARP protein of the 1 x 10^3 μmol/L NaF group. [ Conclusion ] The signal transduction mediated by Caspase plays an important role in the apoptosis induced by high eoncentration of sodium fluoride.

关 键 词：氟化钠 MC3T3-E1细胞 细胞凋亡 CASPASE-3 CASPASE-8 

分 类 号：R994.6[医药卫生—毒理学]