氟对SH-SY5Y细胞活性氧水平和线粒体融合影响  

Changes of ROS level and mitochondria fission-fusion in SH-SY5Y cells with chronic fluorosis

作　　者：楼迪栋[1,2] 郑丹[3] 潘际刚[1] 张凯琳[1] 罗亚[1,2] 于燕妮[1] 官志忠[1,4] 

机构地区：[1]贵阳医学院病理学教研室,贵州贵阳550000 [2]贵阳医学院法医学系 [3]贵阳市妇幼保健院 [4]贵州省分子生物学重点实验室

出　　处：《中国公共卫生》2014年第6期755-757,共3页Chinese Journal of Public Health

基　　金：国家科技支撑计划(2013BAI05B03);国家自然科学基金(81160335);贵州省科学技术基金(黔科合J字(2013)2036号);温州市科学技术基金(Y20080099)

摘　　要：目的观察氟对人脑神经母细胞瘤细胞株(SH-SY5Y细胞)体外培养细胞活性氧水平和线粒体融合的影响。方法在SH-SY5Y细胞中加入0.4、4 mmol/L NaF,孵育6、12、24和48 h;利用MitoTracker和ROS荧光染色试剂盒对线粒体和氧自由基进行染色。结果 NaF作用6、12、24、48 h时,与对照组比较,0.4、4 mmol/L氟处理组SH-SY5Y细胞内ROS染色强阳性细胞计数[分别为(15.1±1.4)、(12.8±2.5)、(22.6±2.4)、(30±3.1)和(32.5±1.9)、(38.1±2.2)、(69.3±3.1)、(84.3±3.5)个/100细胞]和II型线粒体细胞计数[分别为(13±1.4)、(15±1.7)、(22.9±1.9)、(23.7±1.6)和(15.2±2.3)、(23.8±1.8)、(29.1±4)、(47.7±3.3)个/100细胞]明显升高(P<0.05)。结论摄入过量的氟导致SH-SY5Y细胞氧自由基水平升高,线粒体呈分裂状,呈时间和剂量效应关系。Objective To investigate the changes of reactive oxygen species （ROS）level and mitochondrial fission- fusion in SH-SY5Y neurons with chronic fluorosis. Methods Neuroblastoma SH-SY5Y cells were exposed to different concentrations of fluoride（0. 04 and 4 mmol/L）in vitro to build cellular model of fluorosis with the exposure time of 6, 12,24, and 48 hours,respectively. Mitochondrial morphology and ROS levels of SH-SY5Y stained by mito-tracker RED probe and ROS fluorescence dyes were observed with fluorescence microscope. Results Compared with untreatment cells,the numbers of ROS stain positive SY-SY5Y cells （0. 4 mmol/L fluoride： 15.1 ± 1.4, 12. 8 ± 2. 5,22. 6±2.4, 30. 0 ±3. 1 ;4 mmol/L fluoride：32. 5 ±1. 9,38. 1 ±2. 2,69.3 ±3. 1,84. 3±3.5）and the numbers of abnormal mitochon- dria in morphology （ 0. 4 mmol/L fluoride ： 13 ±1. 4,15 ± 1.7,22.9 ±1.9,23.7 ±1. 6 ;4 mmol/L fluoride ： 15.2 ± 2. 3, 23.8 ±1.8,29. 1 ± 4,47.7 ± 3.3 ） were significantly increased after exposure of 6,12,24, and 48 hours （ P 〈 0. 05 ）. Conclusion Taking excessive amount of fluoride results in high level of oxidative stress and mitochondrial fragmentation, which are dependent on the exposure time and dose of fluoride.

关 键 词：氟化物中毒 活性氧 线粒体 SH—SY5Y细胞 

分 类 号：R745.1[医药卫生—神经病学与精神病学]