为探讨氟化钠对小鼠成骨细胞中核因子-κB受体活化因子配体(RANKL)和骨保护素(OPG)mRNA表达比率的影响,选取新生小鼠颅盖骨,用胰蛋白酶-胶原酶消化法分离培养成骨细胞。取其第3代成骨细胞,在实验组培养液中加入不同浓度的氟化钠(10^-7、10^-6和10^-5mol/L),并分别培养24、48、96和144 h,从细胞板贴壁细胞中提取总RNA,应用荧光定量RT-PCR方法检测细胞中RANKL与OPG mRNA表达比率的变化。结果显示,氟化钠增加了RANKL/OPG mRNA的表达比率,且呈明显的时间剂量效应关系。对照组和实验组在培养48 h时,RANKL与OPGmRNA的表达比率显著增加(P〈0.01)。本实验浓度的氟化钠能够增加小鼠颅盖骨成骨细胞中RANKL/OPG mRNA的表达比率。
The objective of this study was to explore the effect of sodium fluoride on the expression of RANKL to OPG mRNA in mouse osteoblasts. Calvariae were derived aseptically from neonatal mouse, and processed by the trypsin and collagenase digestion method. The third-generation cultured osteoblasts were exposed to different concentrations of sodium fluoride (0, 10^-7 , 10^-6 , 10^-5 mol/L) for 24, 48, 96 and 144 h, respectively. The total RNA was extracted, and the change from RANKL to OPG mRNA was observed by real-time RT-PCR. The ratio of RANKL to OPG mRNA expression was up-regulated significantly in mouse osteoblasts in time-dependent and dose-dependent manners after treatment with sodium fluoride. The ratio of RANKL to OPG mRNA for 48 h increased significantly with or with sodium fluoride (P 〈 0.01). The results indicated that sodium fluoride could increase the ratio of RANKL to OPG mRNA in mouse calvarial osteoblasts.