中文摘要：

目的探讨氟对体外培养人成骨细胞中TM9SF1、RasmRNA表达的影响。方法采用原代细胞培养的方法培养人成骨细胞。取早期引产胎儿的骨组织，胶原酶消化、分离成骨细胞，加入DMEM培养液进行传代培养。将传代后的人成骨细胞按染氟剂量不同分为0（对照）、2．5、5．0、10．0、20．0mg／L组。染氟培养10d后，光镜下观察氟对人成骨细胞形态的影响；实时定量（RT）PCR法检测不同染氟剂量对体外培养人成骨细胞TM9SFI、Ras mRNA的表达影响。结果光镜下对照组和2．5mg／L组人成骨细胞呈长梭形、三角形或不规则多边形，有突起伸出，胞质透亮、饱满，相邻细胞彼此贴靠；20．0mg／L组人成骨细胞呈长梭形或不规则多边形，细胞数目明显减少、体积变大，一些细胞胞质出现了多个小空泡及颗粒样物质。不同染氟剂量对人成骨细胞TM9SF1 mRNA表达的影响，组间比较差异有统计学意义（F=322．82，P〈0．01）；其中2．5mg／L组（9326．0＋115．97）表达最高，随着染氟剂量增加，TM9SFImRNA表达降低，5．0、10．0、20．0mg／L组（6495．0±323．9、4387．5＋545．2、5962．5＋536．7）与对照组（9221．0＋107．5）比较，差异有统计学意义（P均〈0．01）。不同染氟剂量对人成骨细胞RasmRNA表达的影响，组间比较差异有统计学意义（F=703．28，P〈0．01）；其中对照组表达最高，随着染毒剂量的增加，RasmRNA表达减少，2．5、5．0、10．0、20．0mg／L组（6144．5±270．82、5603．5＋88．39、3181．0±159．81、4067．5±37．4）与对照组（6571．0±196．58）比较，差异有统计学意义（P均〈0．01）。结论氟影响人成骨细胞TM9SF1、RasmRNA的表达，表达量的高低与染氟剂量有关。

英文摘要：

Objective To detect the influence of fluoride on the expression TM9SF1 mRNA and Ras mRNA of osteoblasts. Methods The third generation of primary cultured osteoblasts were exposed to a series concentrations of 0,2.5,5.0, 10.0,20.0 mg/L fluoride for 10 days. The influence of different doses of fluorine on the expression of TM9SF1 mRNA and Ras mRNA of osteoblasts cultured in vitro was investigated by SYBR Green Ⅰ methods. Results The osteoblasts of the control group and the 2.5 mg/L group were in the shape of long spindle, triangle or irregular polygon and had processes, and the cytoplasm was translucent, adjacent cells affixed to each other under light microscope. Those of the 20.0 mg/L group shaped as long spindle or irregular polygon, and some vacuolization and granular materials appeared in cytoplasm. The number of the cells decreased and the volume increased significantly. After exposed to fluoride for 10 days, osteoblasts of 2.5 mg/L group morphologically proliferated. There were statistical significances between each groups of TM9SF1 mRNA in human osteoblasts（F = 322.82, P 〈 0.01 ）. The highest in the 2.5 mg/L group（9326.0 ± 115.97）, the expression of TM9SF1 mRNA decreased along with the increasing dose of fluorine. There were statistical significances between 5.0,10.0,20.0 mg/L groups（6495.0 ± 323.9, 4387.5 ± 545.2, 5962.5 ± 536.7） and control group（9221.0 ± 107.5, all P 〈 0.01）. There was a statistical significance between each groups of Ras mRNA in human osteoblasts（F = 703.28, P 〈 0.01 ）. The highest in the control group, the expression of Ras mRNA decreased along with the increasing of dose of fluorine. There were statistical significance between 2.5, 5.0, 10.0, 20.0 mg/L groups（6144.5 ± 270.82,5603.5 ± 88.39,3181.0 ± 159.81,4067.5 ± 37.4） and control group （ 6571.0 ± 196.58）. Conclusion The influence on TM9SF1 mRNA and Ras mRNA expression in osteoblasts correlates with the dose of fluorine.