过量氟对大鼠HAT-7细胞系自噬的影响

上海口腔医学 ›› 2016, Vol. 25 ›› Issue (4): 426-430.

过量氟对大鼠HAT-7细胞系自噬的影响

雷双¹, 张颖^{2, 3}, 张凯强², 李健²

1.中国医科大学口腔医学院儿童口腔医学教研室,辽宁省口腔疾病重点实验室, 2.口腔预防医学教研室,辽宁沈阳 110002; 3.上海市口腔病防治院,上海 200031

收稿日期:2015-10-09 出版日期:2016-08-25 发布日期:2016-09-06
通讯作者: 张颖,E-mail：zhangyingcmu@vip.163.com
作者简介:雷双（1990-）,女,硕士研究生,

In vivo and in vitro experimental study on the effect of fluoride-induced autophagy in rat HAT-7 cell line

LEI Shuang¹, ZHANG Ying^{2, 3}, ZHANG Kai-qiang², LI Jian²

1.Department of Pediatric Dentistry, 2.Department of Preventive Dentistry, School of Stomatology, China Medical University, Key Laboratory of Oral Diseases in Liaoning Province.Shenyang 110002, Liaoning Province;
3.Shanghai Stomatological Hospital. Shanghai 200031,China

Received:2015-10-09 Online:2016-08-25 Published:2016-09-06

摘要/Abstract

摘要： 目的研究过量氟对大鼠HAT-7细胞系自噬的影响。方法取大鼠HAT-7细胞,分别加入不同浓度的氟化钠培养液,培养48 h后,透射电子显微镜检测过量氟对大鼠成釉细胞自噬泡的影响,Western 免疫印迹和定量反转录聚合酶链反应技术检测过量氟诱导大鼠成釉细胞内微管相关蛋白轻链（LC3）和Beclin 1 表达的变化。选择40只Wistar大鼠,随机分为A、B、C、D 4组,进行免疫组织化学实验,检测饮水氟对大鼠自噬分子表达的影响。采用SPSS13.0软件包对数据进行统计学分析。结果实验组自噬泡明显增多;Western免疫印迹及定量反转录聚合酶链反应结果显示,随着氟浓度的增加,HAT-7细胞自噬相关基因LC3和Beclin 1表达增加。回归分析结果显示,氟化钠与LC3及Beclin 1的表达有线性关系;大鼠体内免疫组织化学染色结果显示,实验组LC3以及Beclin 1为棕褐色,呈阳性表达。结论过量氟诱导大鼠HAT-7细胞系自噬。

关键词: 氟, 成釉细胞, 自噬, LC3, Beclin 1

Abstract: PURPOSE: To study the effect of fluoride on autophagy in rat ameloblasts both in vitro and in vivo. METHODS: Logarithmic-phase HAT-7 cells were cultured in different concentrations of fluoride for 48h. Transmission electron microscopy (TEM) was used to detect autophagosomes. Western blot and RT-qPCR were carried out to examine the expression of LC3 and Beclin 1. Forty Wistar rats were divided into 4 groups randomly. The expression of LC3 and Beclin 1 in rats was investigated by immunohistochemical staining in vivo. The data were analysed using SPSS13.0 software package. RESULTS: The amount of autophagosomes in the experimental group was significantly more than that in the control group (P<0.05). The expression of LC3 and Beclin 1 were up-regulated in dose dependent manner after treatment with fluoride. Regression analysis showed that fluoride dependently induced the expression of LC3 and Beclin 1 (P<0.05). Immunohistochemical analysis revealed that the expression of LC3 and Beclin 1 in rat ameloblasts in the experimental groups was positive compared to control group. CONCLUSIONS: Excessive fluoride induced autophagy in ameloblasts both in vitro and in vivo.

Key words: Fluoride, Ameloblasts, Autophagy, LC3, Beclin 1

中图分类号:

R780.1

雷双, 张颖, 张凯强, 李健. 过量氟对大鼠HAT-7细胞系自噬的影响[J]. 上海口腔医学, 2016, 25(4): 426-430.

LEI Shuang, ZHANG Ying, ZHANG Kai-qiang, LI Jian. In vivo and in vitro experimental study on the effect of fluoride-induced autophagy in rat HAT-7 cell line[J]. Shanghai Journal of Stomatology, 2016, 25(4): 426-430.

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