摘要
[目的]观察氟中毒大鼠骨组织中Notch3、Jag1蛋白表达水平,探讨其在氟骨症发生机制中的作用。[方法]SD大鼠24只随机分为对照组、低氟组、高氟组3组,每组8只。对照组自由饮用自来水,低氟组、高氟组分别饮用含50、100 mg/L氟化钠的自来水。饲养6个月后处死大鼠。用氟离子选择电极法分别测定尿氟和骨氟,观察骨组织病理学改变,免疫组化检测成骨细胞Notch3、Jag1蛋白表达,免疫印迹法检测骨组织Notch3、Jag1蛋白表达,采用实时荧光定量PCR法检测骨组织Notch3、Jag1 mRNA表达水平。[结果]与对照组相比较,低氟组、高氟组大鼠尿氟、骨氟水平明显增高(均P<0.05)。光镜观察染氟组大鼠出现骨硬化。高氟组大鼠骨组织Notch3蛋白水平与mRNA相对表达含量明显低于对照组(均P<0.05);高氟组大鼠骨组织Jag1蛋白水平和mRNA相对表达含量明显低于对照组、低氟组(均P<0.05)。免疫组化法检测显示低氟组和高氟组大鼠成骨细胞Notch3、Jag1蛋白水平均明显低于对照组(均P<0.05)。[结论]过量氟抑制骨组织特别是成骨细胞Notch3、Jag1表达,使成骨作用增强,该过程可能参与氟骨症发病机制。
[ Objective ] To explore the expressions of Notch3 and Jagl proteins in bone tissues of fluorosis rats and their roles in the pathogenesis of skeletal fluorosis. [ Methods ] Twenty-four SD rats were randomly divided into control, low-fluoride, and high-fluoride groups, eight rats each. The three groups were fed with tap water and water containing 50 mg/L and 100 mg/L sodium fluoride, respectively. All rats were neutralized after six months. Fluoride levels in urine and bone were measured by fluoride ion selective electrode method. Bone tissue samples were observed for pathological changes. The protein levels of Notch3 and Jagl in osteoblasts were detected by immunohistochemical assay. The expressions of Notch3 and Jagl proteins and mRNA in bone tissues were detected by Western blot and real-time fluorescent quantitative PCR respectively. [ Results] Compared with the control group, the fluoride levels in urine and bone in the low- and high-fluoride groups were significant higher (all Ps 〈 0.05). Osteosclerosis was observed under optical microscope. The protein and mRNA expression levels of Notch3 in bone tissues of the high-fluoride group were significant lower than those of controls (both Ps 〈 0.05), meanwhile the protein and mRNA expression levels of Jagl in bone tissues of the high-fluoride group were significant lower than those of the control and low-fluoride groups (all Ps 〈 0.05). The protein expression levels of Notch3 and Jag1 in osteoblast of the low- and high-fluoride groups were significant lower than those of the controls (all Ps〈0.05) by immunohistochemical assay. [ Conclusion ] Excessive fluoride could inhibit the expressions of Notch3 and Jagl in bone tissues, especially in osteoblasts, and enhance osteogenesis as well, which might be connected to the pathogenesis of skeletal fluorosis.
出处
《环境与职业医学》
CAS
CSCD
北大核心
2016年第5期494-498,共5页
Journal of Environmental and Occupational Medicine
基金
贵州省科技厅社会发展攻关资金(编号:黔科合LG字[2012]056号)