[氟对人神经母细胞瘤SH-SY5Y细胞自噬水平的影响]

[Effects of fluoride on autophagy level in human neuroblastoma SH-SY5Y cells].

作者信息

Tang Sha, Zhang Shun, Chen Wei, Quan Chao, Duan Peng, Huang Wenting, Wang Aiguo, Yang Kedi

机构信息

Department of Occupational and Environmental Health, School of Public Health, MOE Key Lab of Environment and Health, Tongji Medical College, Huazhong University of Science and Technology Wuhan 430030, China.

出版信息

Wei Sheng Yan Jiu. 2017 May;46(3):472-480.

PMID:29903262

Abstract

OBJECTIVE

To explore the effects of fluoride on autophagy level in human neuroblastoma SH-SY5Y cells.

METHODS

SH-SY5Y cells were treated with different concentrations( 20, 40, 60 mg/L) of fluoride for 24 h. Transmission electron microscope( TEM) was used to detect the ultrastructure of autophagosomes in SH-SY5Y cells. Acridine orange staining was used to examine the form of autophagic vesicles and Western blotting was used to check the expression levels of autophagy relevant proteins of Atg5, LC3 and P62 in SH-SY5 Y cells.

RESULTS

The TEM results showed the presence of autophagic ultrastructure in SH-SY5Y cells and fluoride could decrease the amount of autophagosomes. Acridine orange staining results showed that the amount of autophagic vesicles were obviously decreased in 40 and 60 mg/L Na F-treated groups compared with control, and 60 mg/L Na F-treated group was decreased the most. The expression level of Atg5 and LC3-II protein was dose-dependently decrease and P62 protein was dosedependently increased compared with control( P < 0. 05).

CONCLUSION

Fluoride maysuppress the level of autophagy in SH-SY5Y cells.

摘要

目的

探讨氟对人神经母细胞瘤SH-SY5Y细胞自噬水平的影响。

方法

用不同浓度（20、40、60mg/L）的氟处理SH-SY5Y细胞24小时。采用透射电子显微镜（TEM）检测SH-SY5Y细胞中自噬体的超微结构。用吖啶橙染色检测自噬泡的形态，并用蛋白质免疫印迹法检测SH-SY5Y细胞中自噬相关蛋白Atg5、LC3和P62的表达水平。

结果

透射电镜结果显示SH-SY5Y细胞中存在自噬超微结构，氟可减少自噬体的数量。吖啶橙染色结果显示，与对照组相比，40和60mg/L氟化钠处理组的自噬泡数量明显减少，其中60mg/L氟化钠处理组减少最多。与对照组相比，Atg5和LC3-II蛋白的表达水平呈剂量依赖性降低，P62蛋白呈剂量依赖性升高（P<0.05）。