万雯,万良斌,于燕妮,万昌武,官志忠
WAN Wen,WAN Liangbin,YU Yanni,WAN Changwu,GUAN Zhizhong(Department of Pathology

• 1:贵阳医学院病理学教研室

摘要(Abstract)：

目的:观察氟对大鼠骨组织中Twist mRNA及其蛋白表达的影响,探讨Twist在氟中毒引起氟骨症发病中的作用。方法:36只SD大鼠按性别和体重随机分为3组,每组12只,组内雌雄各半;对照组自由饮用自来水,低氟组、高氟组分别饮用含氟水5 mg/L、50 mg/L;实验6个月后股动脉放血处死大鼠,采用氟离子选择电极法检测大鼠尿氟、骨氟含量;HE切片观察大鼠股骨下段骨组织病理学改变,测定骨皮质厚度、骨小梁宽度等骨形态学变化;采用原位杂交技术及免疫组织化学方法检测慢性氟中毒大鼠Twist mRNA及其蛋白表达水平。结果:与对照组比较,氟中毒组大鼠尿氟及骨氟含量明显升高,股骨远端骨皮质增厚、骨小梁数目增多、骨髓腔间隙变小等骨质硬化表现,形态学计量指标的差异有统计学意义(P<0.05),且随染氟剂量增加,以上病理改变逐渐加重;与对照组比较,染氟组Twist mRNA及其蛋白表达水平明显降低,差异有统计学意义(P<0.05)。结论:氟可导致大鼠骨组织中Twist mRNA及其蛋白表达水平降低,Twist可能参与氟中毒引起的骨骼损伤。
Objective: To observe the effect of fluoride on the expressions of Twist mRNA and protein in bone tissue of rats,and to explore the role of Twist in pathogenesis of skeletal fluorosis.Methods: Thirty-six SD rats were randomly divided into 3 groups with 12 rats in each and half male half female.Controls were fed with tap water,rats of low fluoride and high fiuoride groups were fed with water containing fluoride 5 mg/L and 50 mg/L respectively to establish fluorosis models.Rats were sacrificed in 6 months by femoral artery exsanguination.Urinary fluoride and bone fluoride contents of rats were detected by fluoride ion-selective electrode method.HE staining was used to observe pathological changes of bone tissue of rat femoral inferior segment,and the morphological changes such as thickness of cortical bone and width of trabecular were detected.Twist protein and mRNA expression levels were detected with immunohistochemistry and in situ hybridization respectively.Results: The fluorine contents of urine and bone in fluorosis groups were higher than those in the control rats.Osteosclerosis manifestations such as thickened cortical bone,increased trabecular number and smaller marrow cavity were found in femoral inferior segment of fluorosis rats,and the differences of these indexes were significant(P<0.05).With the increase of fluoride contents,the pathological changes mentioned above were more serious.Twist mRNA and protein expression levels of fluorosis groups were lower than those of the controls,the difference was statistically significant(P<0.05).Conclusions: Fluoride can reduce the expression levels of Twist mRNA and protein in bone tissue of rats.Twist may play a role in bone damage caused by fluorosis.

关键词(KeyWords)： 氟;中毒;Twist;原位杂交;免疫组织化学;骨
fluorine;poisoning;Twist;in situ hybridization;Immunohistochemistry;bone

Abstract:

Keywords:

基金项目(Foundation): 国家科技部国际合作重点项目(2010DFB30530);; 贵州省科技厅社会发展攻关项目[黔科合SY字(2011)3006号]

作者(Author): 万雯,万良斌,于燕妮,万昌武,官志忠
WAN Wen,WAN Liangbin,YU Yanni,WAN Changwu,GUAN Zhizhong(Department of Pathology

DOI: 10.19367/j.cnki.1000-2707.2012.03.002

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