氟化钠对大鼠成骨肉瘤细胞增殖活性及碱性磷酸酶活力和骨钙素mRNA表达的影响  

Effects of sodium fluoride exposure on cell viability and ALP activity and BGP mRNA expression in rat osteosarcoma UMR-106 cells

作　　者：王楠兰 敖全 雷露[1] 曾贝贝[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：《环境与健康杂志》2016年第9期773-776,共4页Journal of Environment and Health

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

摘　　要：目的 探讨氟化钠（Na F）对成骨样细胞大鼠骨肉瘤（UMR-106）细胞增殖活性及主要成骨活性标志物碱性磷酸酶（ALP）活力和骨钙素（BGP）m RNA表达的影响。方法 将处于对数生长期的UMR-106细胞分别暴露于含终浓度为0（对照）、5×10、102、5×102、1×103、5×103、1×104、2×104、4×104、8×104μmol/L Na F的DMEM培养液中孵育24、48 h,采用CCK-8检测细胞活性。将处于对数生长期的UMR-106细胞分别暴露于含终浓度为0（对照）、1×103、2×103、4×103μmol/L Na F的DMEM培养液（不含FBS）继续培养12、24、48 h,测定细胞培养液和细胞ALP的活力及细胞中BGP m RNA的表达水平。结果 与对照组比较,1×103~8×104μmol/L Na F染毒24 h和5×10~8×104μmol/L Na F染毒48 h时大鼠UMR-106细胞的存活率均较低,差异有统计学意义（P〈0.05）。5×10、1×103、5×103、1×104、2×104μmol/L Na F染毒48 h时大鼠UMR-106细胞的存活率均低于染毒24 h时,差异有统计学意义（P〈0.05）。各浓度Na F染毒组UMR-106细胞培养液和细胞中ALP活力均较对照组高,除1×103μmol/L Na F染毒24 h的细胞培养液和1×103μmol/L Na F染毒48 h的细胞内ALP活力外,差异有统计学意义（P〈0.05）。随着Na F染毒浓度的升高,UMR-106细胞培养液和细胞中ALP活力均呈上升趋势;随着Na F染毒时间的延长,细胞培养液中ALP的活力呈上升趋势,而细胞中ALP的活力呈先升高后下降的趋势。2×103、4×103μmol/L Na F染毒组UMR-106细胞的BGP m RNA表达水平均较对照组高,差异有统计学意义（P〈0.05）。随着Na F染毒浓度的升高,UMR-106细胞BGP m RNA的表达水平呈上升趋势;随着Na F染毒时间的延长,UMR-106细胞BGP m RNA的表达水平呈下降趋势。结论 在本实验浓度下,Na F对UMR-106细胞的增殖呈抑制作用,但可促进其成骨活性标志物ALP活力及BGP m RNA的表达。Abstract：Objective To explore the effects of sodium fluoride （NaF） exposure on cell viability activity and biomarker of osteogenic activity expression in rat osteosarcoma UMR-106 cells. Methods UMR-106 cells were exposed to 0 （control）,5×10, 102, 5 × 102,1 × 103, 5 × 103,1 × 104, 2 ×104, 4 ×104 and 8 ×104 μ mol/L NaF solution for 24 h and 48 h. CCK-8 assay was used to measure cell viability. After UMR-106 cells were treated with 0 （control）, 103,2×103,4×103 μmol/L NaF for 12 h,24 h and 48 h, ALP activity and osterocalein mRNA were examined by the methods of disodium phenyl phosphate and real-time PCR. Results Compared with the control, cell viability significantly decreased in 1 ×103-8×104 μ mol/L sodium fluoride treatment groups for 24 h and 5×10-8×104 μ mol/L sodium fluoride treatment groups for 48 h （P〈0.05）. And the cell viabilities in 5×10, 1×103,5×103,1×1042×104 μ mol/L NaF treatment groups for 48 h were significantly lower than 24 h （P〈0.05）. At different NaF concentrations, ALP activities of the cell and the cell culture supematants were higher than those of control, except the cell and the cell culture supernatants in 1×103 μmol/L NaF treatment group for 24 h （P〈0.05）. With the increase of NaF concentration, the ALP activities of the cell and the cell culture supernatants were showed increasing trend ;With the prolongation of exposure time,the ALP activities of the cell culture supernatants were increasing and the cell showed increasing trend first,then decreasing （P〈0.05）. Then the BGP mRNA of the cell in 2×103,4×103 μ mol/L NaF treatment groups showed increasing trend. With the prolongation of exposure time,the BGP mRNA of the cell showed decreasing trend. Conclusion NaF can inhibit osteoblastie cell viability and can also promote the expression of osteogenic activity biomarker.

关 键 词：氟化钠 成骨肉瘤细胞 增殖活性 碱性磷酸酶 骨钙素 

分 类 号：R994.6[医药卫生—毒理学]