氟对原代培养的大鼠成骨细胞分化相关指标的影响  

Effects of fluoride on osteoblastic differentiation-related indexes in primary cultured rat osteoblasts

作　　者：李晓娜[1] 潘磊磊[2] 郭晓英[2] 

机构地区：[1]中国医科大学基础医学院,辽宁沈阳110001 [2]中国医科大学公共卫生学院地球化学性疾病研究室

出　　处：《环境与健康杂志》2014年第2期118-120,F0003,共4页Journal of Environment and Health

基　　金：辽宁省科学技术计划项目(2013225049)

摘　　要：目的研究氟对原代培养的大鼠成骨细胞的细胞活性及对成骨细胞分化相关指标的影响。方法取出生24h内的大鼠胎鼠的颅盖骨，采用胰酶和Ⅱ型胶原酶交替消化的方法获得原代成骨细胞。分别加入终浓度为0（对照组）、10^-7、10。、10^-5、10^-4mol／L氟化钠进行染毒72h，采用MTT法检测细胞活性；染氟72h后，采用Real—time RT—PCR法检测成骨细胞分化标志物COL1A1、ALP和ON mRNA的表达情况；染氟5d后，采用染色法检测成骨细胞ALP蛋白的表达情况。结果与对照组相比，10^-6～10^-4mol／L染氟组成骨细胞存活率及10^-7-10^-5mol／L染氟组成骨细胞ALP蛋白的表达水平均较高，差异有统计学意义（P〈0．05，P〈0．01）；且随着NaF染毒浓度的升高，成骨细胞存活率和ALP蛋白的表达水平均呈先升高后下降的趋势。与对照组相比，各浓度NaF染毒组成骨细胞ALP、ONmRNA的表达水平及10^-7、10^-6mol／LNaF染毒组成骨细胞COL1A1 mRNA的表达水平均升高，差异有统计学意义（P〈0．05，P〈0．01）。结论本实验条件下，较低剂量的氟可促进大鼠成骨细胞的分化。Objective To study the effects of fluoride on cell viability and mRNA expressions of osteoblast differentiation-related indexes in primary cultured rat osteoblasts. Methods The osteoblasts derived from calvaria of new-born Wistar rats were isolated by trypsin and type Ⅱ collagenase digestion. Cell viability was examined by MTT assay,mRNA expressions of COL1A1 ,ALP and ON were evaluated by real-time RT-PCR after treatment with various concentrations of fluoride （0,10^-7,10^-6, 10^-5 and 10^-4 mol/L） for 72 h. Protein expression of ALP was examined by staining kit after treatment with various concentrations of fluoride （0,10^-7,10^-6,10^-5 and 10^-4 mol/L） for 5 d. Results Compared with the control, cell viability significantly increased by 10^-6-10^-4 mol/L sodium fluoride （P〈0.05,P〈0.01）. Expression of ALP protein was significantly increased after 10^-7-10^-5 mol/L fluoride treatment for 72 h in osteoblasts compared with the control cell （P〈0.05,P〈0.01）. Compared with the control, expression of ALP, ON mRNA in fluoride-treated groups and expression of COL1A1 mRNA in 10^-7, 10^-6 mol/L NaF-treated group significantly increased （P〈0.05,P〈0.01）. Conclusion Low concentration of fluoride may induce osteoblastie differentiation in rats.

关 键 词：氟 成骨细胞 原代培养 分化 

分 类 号：R994.6[医药卫生—毒理学]