›› 2020, Vol. 40 ›› Issue (11): 982-986.

• 基础研究 • 上一篇    下一篇

自噬在氟牙症大鼠成釉细胞中的保护作用

翁清清,易芳羽,郁莹,葛苏钰,张颖   

  1. 上海市口腔病防治院
  • 收稿日期:2020-03-06 修回日期:2020-04-22 出版日期:2020-11-28 发布日期:2020-11-24
  • 通讯作者: 张颖 E-mail:zhangyingcmu@vip.163.com
  • 基金资助:
    国家自然科学基金;上海市自然科学基金

Protect effect of autophagy on the ameloblast apoptosis in rats with chronic fluorosis

  • Received:2020-03-06 Revised:2020-04-22 Online:2020-11-28 Published:2020-11-24
  • Contact: Ying -ZHANG E-mail:zhangyingcmu@vip.163.com

摘要: 目的 研究不同程度氟牙症大鼠模型切牙成釉细胞的自噬及凋亡水平,探讨自噬在氟牙症大鼠成釉细胞凋亡中的作用。方法 48只4周龄Wistar大鼠随机分为6组,通过饮水加75 mg/L和150 mg/L氟离子和腹腔注射10 mg/kg 3-甲基腺嘌呤(3-methyladenine,3-MA)的方法建立氟牙症大鼠模型及自噬抑制大鼠模型。观察成釉细胞形态改变和自噬相关蛋白Beclin1、自噬底物蛋白p62、微管相关蛋白轻链3(microtubule-associated protein light chain 3,LC3)及活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)蛋白表达水平,采用SPSS 22.0软件包对免疫组化定量结果进行统计学分析。结果 随着饮水氟离子含量增加,切牙氟牙症表现明显,成釉细胞形态结构紊乱。免疫组化结果显示饮用150 mg/L含氟水的大鼠成釉细胞中Beclin1、LC3和Cleaved Caspase-3蛋白表达量增加,p62表达量下降,相较饮用75 mg/L含氟水组大鼠改变更加明显。同时,在相同饮水条件下,3-MA抑制自噬后成釉细胞中Beclin1和LC3表达量下降,p62表达量升高,Cleaved Caspase-3蛋白水平明显增加。结论 氟牙症大鼠成釉细胞中存在自噬,高浓度氟刺激对其诱导作用加强,并且自噬在一定程度上可以保护氟牙症大鼠成釉细胞免受凋亡。

关键词: 氟, 成釉细胞, 自噬, 凋亡

Abstract: Objective To investigate the level of autophagy and apoptosis of ameloblasts in rats with different degrees of fluorosis, and to explore the effect of autophagy on the ameloblast apoptosis in rats with chronic fluorosis. Methods Forty-eight Wistar rats of 4-week-olds were randomly divided into 6 groups to establish a fluorosis rat model by adding 75 mg/L or 150 mg/L fluoride to the drinking water, and an autophagy inhibition model treated with an intraperitoneal injection of 10 mg/kg 3-methyladenine (3-MA). The morphology and expression of Beclin1, p62, LC3, and Cleaved Caspase-3 in rat incisor ameloblasts were observed, and statistical analysis of quantitative results of immunohistochemistry was performed using SPSS 22.0 software package. Results With the increase of fluoride in drinki ng water, features of dental fluorosis became more obvious and found disorders in morphological and structural of ameloblasts. Results of immunohistochemistry showed that the expression of Beclin1 and LC3 increased while p62 expression decreased in rat ameloblasts drinking 150 mg/L fluoride, and the expression of Cleaved Caspase-3 increased. these changes are more obvious than rats drinking 75 mg/L fluoride-containing water. After administration 3-MA to inhibit autophagy, and they, in turn, decreased the expression of Beclin1 and LC3 and increased the p62 expression. Meanwhile, the expression level of Cleaved Caspase-3 in ameloblast increased significantly. Conclusions There is autophagy in rat ameloblast with chronic fluorosis, the induction of it is enhanced by the high concentration of fluoride and may play a protective role in reducing the level of apoptosis in ameloblasts during the fluorosis.

Key words: Fluoride, Ameloblast, Autophagy, Apoptosis

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