氟对体外培养大鼠成釉细胞活性和细胞凋亡的影响

上海口腔医学 ›› 2014, Vol. 23 ›› Issue (5): 519-523.

氟对体外培养大鼠成釉细胞活性和细胞凋亡的影响

马林¹, 张颖¹, 钟鸣², 朱莉², 张凯强¹, 顾何锋¹, 刘璐¹, 张思宇¹, 程睿波¹

1.中国医科大学口腔医学院口腔预防教研室; 2.中心实验室,辽宁省口腔医学研究所,辽宁沈阳 110002

收稿日期:2013-10-21 修回日期:2013-12-10 出版日期:2014-10-20 发布日期:2015-02-04
通讯作者: 张颖,E-mail: zhangyingcmu@vip.163.com
作者简介:马林（1986-）,女,硕士,医师,E-mail：jrma6143@sina.com
基金资助:
国家自然科学基金（81072245）; 辽宁省自然科学基金（20102278）

Effect of fluoride on the viability and apoptosis of ameloblasts in vitro

MA Lin¹, ZHANG Ying¹, ZHONG Ming², ZHU Li², ZHANG Kai-qiang¹, GU He-feng¹, LIU Lu¹, ZHANG Si-yu¹, CHENG Rui-bo¹

1.Department of Preventive Dentistry; 2.Department of Central Laboratory, School of Stomatology, China Medical University. Shenyang 110002, Liaoning Province, China

Received:2013-10-21 Revised:2013-12-10 Online:2014-10-20 Published:2015-02-04
Supported by:
; Supported by National Natural Science Foundation of China (81072245) and Natural Science Foundation of Liaoning Province (20102278)

摘要/Abstract

摘要： 目的观察氟对体外培养的大鼠成釉细胞活性的影响,为探讨氟斑牙的形成机制提供依据。方法取对数生长期的大鼠成釉细胞,在培养液中加入浓度为0、0.4、0.8、1.6、3.2、6.4 mmol/L的 NaF溶液,培养24、48、72 h后,采用CCK-8检测各组细胞的活性。荧光显微镜下观察成釉细胞核形态的变化,流式细胞术分析氟对细胞凋亡的影响。采用SPSS13.0软件包对数据进行统计学分析。结果 ①NaF浓度为0.4、0.8 mmol/L时,对成釉细胞有促增殖作用;NaF浓度为1.6、3.2、6.4 mmol/L时,对成釉细胞的活性有抑制作用,随着NaF浓度的增加,对细胞的抑制作用也逐渐增强,并且这种双向调节作用呈时间依赖性。②NaF浓度为0.4、0.8 mmol/L时,鲜见核破碎;NaF浓度为1.6、3.2 mmol/L时,存在核破碎,并且随着浓度的提高,核破碎的数量随之增加,即1.6 mmol/L浓度的NaF可引起成釉细胞凋亡,随着浓度的增加,细胞凋亡的数量随之增加。结论 ①氟对体外培养成釉细胞的增殖具有双向调节作用,即低浓度促进,高浓度抑制。②浓度超过1.6 mmol/L时,NaF诱导成釉细胞凋亡。

关键词: 氟, 成釉细胞, CCK8, 流式细胞术, Hoechst 33258荧光染色

Abstract: PURPOSE: To evaluate the effect of fluoride on viability of rat ameloblasts in vitro. METHODS: The ameloblasts of rat was exposed to different concentrations of NaF (0, 0.4, 0.8, 1.6, 3.2, 6.4 mmol/L) for 24, 48 and 72 hours. CCK-8 assays were performed to measure the cells proliferation; The morphology of apoptosis was observed by Hoechst 33258 staining and the rate of apoptosis was determined by flow cytometry. The data was analyzed using SPSS 13.0 software package. RESULTS: ①The proliferation of ameloblasts was increased when concentrations of NaF between 0.4 mmol/L and 0.8 mmol/L, whereas inhibited at 1.6 mmol/L NaF and above. The effects were in time-dependent manner.②Cells in the 1.6 mmol/L NaF groups showed unclear karyorrhexis and apoptotic cell morphology. The effects were in concentration-dependent manner. CONCLUSIONS: ① Fluoride has two-phase effects to ameloblasts: At low doses, it promoted cell proliferation while at high doses it had negative effects. ②1.6 mmol/L NaF could induce apoptosis of ameloblasts.

Key words: Fluoride, Ameloblast, CCK8, Flow cytometry, Hoechst 33258 fluorescence staining

马林, 张颖, 钟鸣, 朱莉, 张凯强, 顾何锋, 刘璐, 张思宇, 程睿波. 氟对体外培养大鼠成釉细胞活性和细胞凋亡的影响[J]. 上海口腔医学, 2014, 23(5): 519-523.

MA Lin, ZHANG Ying, ZHONG Ming, ZHU Li, ZHANG Kai-qiang, GU He-feng, LIU Lu, ZHANG Si-yu, CHENG Rui-bo. Effect of fluoride on the viability and apoptosis of ameloblasts in vitro[J]. Shanghai Journal of Stomatology, 2014, 23(5): 519-523.

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