氟化钠对小鼠成骨细胞MC3T3-E1凋亡的影响

发布时间：2019-01-08 14:05

【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(P0.05)。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(P0.05)。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(P0.05)。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(P0.05)。[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。
[Abstract]:[objective] to investigate the effect of high concentration sodium fluoride (Na F) on apoptosis of mouse osteoblast MC3T3-E1 cells. [methods] MC3T3-E1 cells were incubated with different doses of Na F for 24 h and 48 h, respectively. The survival rate of MC3T3-E1 cells was determined by CCK-8 assay. The concentration and time of the subsequent experiment were determined. The concentration and time of the subsequent experiment were determined. 0 脳 10 ~ 3 脳 10 ~ 3 渭 mol/L Na F ~ 2 脳 10 ~ (3) C ~ (3) 脳 10 ~ (3) 渭 mol/L Na F in mouse osteoblasts MC3T3-E1,. Apoptosis was detected by flow cytometry (FCM) after cultured for 48 h. The expression of Caspase-3 and Caspase-8 m RNA was detected by real time PCR and the expression of Cleaved Caspase-3,Cleaved PARP protein was detected by Western blot. [results] CCK-8 results showed that with the increase of Na F concentration, the survival rate of MC3T3-E1 cells decreased with the increase of Na F concentration and the prolongation of exposure time. The OD value of 1 脳 10 ~ (-3) 渭 mol/L was lower than that of the control group, and the survival rate was decreased. The difference was statistically significant (P0.05). The OD value of 1 脳 10 3 渭 mol/L Na F at 48 h was lower than that of 24 h (P0.05). Compared with the control group for 24 h, the apoptosis rate of 1 脳 10 ~ 3 渭 mol/L Na F group was not changed significantly, but the apoptosis rate of other Na F groups was higher than that of the control group (P0.05). With the increase of Na F concentration, the expression of Caspase-3,Caspase-8 m RNA in MC3T3-E1 cells increased. After 48 h of Na F exposure, the expression of Cleaved Caspase-3,Cleaved PARP protein in MC3T3-E1 cells increased except the Cleaved PARP protein expression of 1 脳 10 3 渭 mol/L Na F group (P0.05). [conclusion] Caspase mediated signal pathway plays an important role in the process of apoptosis induced by high concentration of Na F.
【作者单位】：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;
【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)
【分类号】：R114

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