氟对体外培养的成骨细胞Ras蛋白表达的影响

Effect of protein expression of Ras gene in osteoblasts cultured in vitro and exposed to fluoride

目的:研究不同浓度的氟化物对体外培养的成骨细胞Ras蛋白表达的影响。方法:采用酶消化法分离乳鼠成骨细胞,经剂量分别为0、2.5、5.0、10.0和20.0mg/L的氟化钠(NaF)作用后,采用免疫组织化学法和Western blot法检测染氟后小鼠成骨细胞中Ras蛋白表达水平。结果:免疫组织化学法结果显示0、2.5及5.0mg/L实验组成骨细胞中Ras均有较强阳性表达,其余实验组的为弱阳性表达;各染氟组阳性表达细胞数与0mg/L比较差异有统计学意义(P<0.05)。小鼠细胞的Ras蛋白相对定量结果显示:2.5、5.0mg/L染氟组大于0mg/L,其它染氟组均小于0mg/L,染氟组的蛋白相对表达量有随着剂量增加而逐渐降低的趋势。结论:氟作用于成骨细胞后可以影响Ras蛋白的表达,表现为双向作用,即低剂量的氟促进Ras的蛋白表达,高剂量抑制Ras的蛋白表达。

Objective： To study effect of protein expression of Ras gene in osteoblasts cultured in vitro and exposed to different does of fluoride. Methods： Osteoblasts were obtained from the calvaria of mice within 1～3 day by enzymic digestion method and were cultured. Then osteoblasts were exposed to different does of fluoride. Detected protein expression of Ras of mice osteoblasts exposed different does of fluoride by immunohistochemistry and Western blotting. Results： All groups had protein postitive expression after exposed to fluoride, but the 2.5 mg/L fluoride groups of postitive degree and area were stonger and bigger than controal group and the rest fluoride groups of postitive degree and area were weaker and smaller than controal group. Calculation of postitive osteobalsts numbers showed that it had statistic difference between the fluoride groups and the controal group. The numbers of 2.5 mg/L fluoride groups were higher than the control group （P d0.05） and the numbers of the rest fluoride groups were lower than the control one （P 〈0.05）. Among of these number of 2.5 mg/L fluoride group was the highest. The correspondence quantitation result of protein of Ras of mice showed that the numbers of 2.5 mg/L and 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 fluoride dose. Conclusions： F～uoride influence the protein expression of Ras of osieoblasts,which shows double action：low does of fluoride improves and high does of fluoride inhibits.