摘要
目的探讨氟对大鼠骨组织中破骨细胞的作用及其调节机制。方法将20只清洁雄性3周龄Wistar大鼠，按体重采用随机数字表法分为两组，每组10只。对照组饮用蒸馏水，染氟组饮用含氟100mg/L的蒸馏水，饲养3个月。观察大鼠氟斑牙情况，离子选择电极法检测骨氟蓄积量，光镜观察大鼠骨组织病理形态变化．抗酒石酸酸性磷酸酶（TRAP）染色法鉴定骨组织中破骨细胞，定磷法检测血清钙调磷酸酶（CaN）活力，二喹啉甲酸（BCA）法检测血清总蛋白浓度，比色法检测血清中钙（Ca）和丙二醛（MDA）含量，酶联免疫吸附法（ELISA）测定钙调蛋白（CaM）含量。结果实验结束时，对照组无氟斑牙检出，染氟组大鼠均有氟斑牙表现；染氟组大鼠骨氟含量[（4460．671±418．548）mg／kg]高于对照组[（582．5344-58．342）mg／kg，t=-29．020，P〈0．01]；与对照组比较，染氟组大鼠骨组织中呈现骨小梁普遍变粗、骨质融合、骨矿化不全等改变；染氟组大鼠骨组织中TRAP染色阳性信号强度均显著高于对照组，染氟组破骨细胞形成数量[10（5～12）个]也显著多于对照组[3（2～4）个，U=92．5，P〈0．01]；染氟组大鼠血清中CaN活力[（3．334±0．654）nmol／mgprot]显著高于对照组[（1．289±0．361）nmol／mgprot，t=-6．346，P〈0．01]，两组大鼠血清中Ca和CaM含量无显著差别，而染氟组MDA含量[（7．703±2．954）[mol／L]显著高于对照组[（3．958±1．965）μmol／L，t=-2．968，P〈0．05]。结论过量氟可导致大鼠骨组织中破骨细胞生成增多，其机制可能为氟通过氧化应激途径刺激机体内CaN活性的升高。
Objective To investigate the effect of fluoride on osteoclast in bone tissue of rats and its mechanism. Methods Twenty specific pathogen free male Wistar rats aged 3 weeks were randomly divided into two groups by weight （each group has 10）. The rats of control group drink distilled water and treatment group drink distilled water containing 100 mg/L fluoride. The rats were fed for 3 month. The dental fluorosis in rats was observed. The ion selective electrode method was used to measure bone fluoride accumulation. The pathological changes of bone tissue in rats were observed under light microscope. The osteoclast was identified by tartrateresistant acid phosphatase （TRAP） staining. The calcineurin （CAN） activity of serum was measured by detection of free phosphate with malachite green. The bicinchoninic acid （BCA） method was used to detect total protein concentration of serum. The colorimetry method was used to detect calcium and malondialdehyde （MDA） levels in serum. The enzyme linked immunosorbent assay （ELISA） method was used to detect calmodulin （CAM） content. Results By the end of the experiment, none dental fluorosis was detected in control group, all rats in fluoride group had dental fluorosis. The bone fluoride content of rats in fluoride group [（4 460.671± 418.548） mg/kg] was about 7.6 times higher than that in contr±ol group [（582.534 ± 58.342） mg/kg, t = - 29.020, P 〈 0.01]. Compared with the control group, the bone tissue of rats in fluoride group showed thicker bone trabecular, sclerotin fusion and incomplete mineralization. Positive signal intensity of TRAP staining of bone tissue in fluoride group was significantly higher than that in control group. The number of osteoclast formation in fluoride group [10 （5 - 12）] was significantly higher than that in control group [3 （2 - 4）; U = 92.5, P 〈 0.01]. CaN activity in serum of rats in fluoride group [（3.334 ± 0.654） nmol/mg prot] was significantly higher than that in control group [（1.289 ± 0.561） nmol/mg prot; t = - 6.346, P 〈 0.01]. The Ca and CaM content of serum in rats were not significantly different between the two groups. However MDA content in fluoride group [（7.703 ± 2.954） μmol/L] was significantly higher than that in control group [（3.958 ±1.965） μmol/L, t = - 2.968, P 〈 0.05]. Conclusion Excessive fluoride may increase osteoclast formation in bone tissue of rats, and the mechanism might be fluoride stimulated CaN activity through oxidative stress pathway.
出处
《中华地方病学杂志》
CSCD
北大核心
2017年第10期714-718,共5页
Chinese Journal of Endemiology
基金
黑龙江省教育厅科学技术研究项目（12541463）
