Effect Of Ras Gene Expression On Osteoblasts Cultured In Vitro After Exposed To Fluoride

Objective: To detect protein and RNA expression of Ras of mice osteoblasts and quantity of RasmRNA of human osteoblasts when osteoblasts was exposed different does of fluoride. Methods: After built and improved the method of osteobalsts primary culture in vitro in mice, we identified the origination of osteobalsts by AKP and calcium nodes dyeing; we detected Ras protein expression level and quantity of RasmRNA of Ras of mice osteoblasts which were exposed to different does of fluoride by immunohistoche-mistryl, hybridization in situ,western blottingting; to detect RasmRNA of human osteoblastsan which were exposed to different does of fluoride by Real Time PCR. Results: The culture method of osteobalsts in mice was repeated well. Cells grew stably and were purified successful which was basement of fluorosis model in vitro. All groups had Ras protein and RNA postitive expression after exposed to fluoride, but t postitive degree and scope in 2.5mg/L fluoride group were stonger and bigger than controal group and other fluoride groups of postitive degree and area were weaker and smaller than control group. Calculation of postitive osteobalsts numbers showed that it had statistic difference between the fluoride group and the control group. The numbers of 2.5mg/L fluoride groups were higher than the control group (p<0.05) and the numbers of the rest fluoride groups were lower than the control one (p<0.05). Among of these, numbers of 2.5mg/L fluoride groups were highest. The correspondence quantitation of RasmRNA in mice showed that the quantity of 2.5mg/L fluoride groups were more than the control group and the numbers of the rest fluoride groups were less than the control one, but the fluoride groups had decreased tendency with increase the the fluoride dose. The correspondence quantitation result of protein of Ras of mice showed that quantity of 2.5mg/Land 5.0mg/L fluoride groups were more than the control group and the numbers of the rest fluoride groups were less than the control one. The quantitation of RasmRNA of the fluoride groups were less the control one. Conclusions: Fluoride influences the Ras protein and mRNA expression of Ras in mice osteoblasts, which shows double action:low does of fluoride improves and high does of fluoride inhibits. Fluoride influences RasmRNA of human osteoblasts, and and the quantity of RasmRNA of human osteoblasts decrease with the increase of fluoride in the experience.