Analytical and Quantitative Cytopathology and Histopathology
2019, Volume 41, Issue 4
Research Article
Autophagy is involved in the apoptosis of human retinal pigment epithelial cells induced by hydrogen peroxide
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1
Departments of Neurology and of Ophthalmology, Xi'an Jiaotong University, Xi'an, Shaanxi, China
2
Department of Ophthalmology, Urumqi Aier Eye Hospital, Urumqi, Xinjiang, China
3
Department of Clinical Medicine, Xi'an Medical University, Xi'an, China
4
Department of Ophthalmology, Xi'an Medical University, Xi'an, China
5
Center for Translational Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi, China
Abstract
OBJECTIVE: To investigate the role of autophagy on the cell viability and apoptosis of human retinal pigment epithelial cells (RPE-19) in the condition of oxidative stress. STUDY DESIGN: RPE-19 cells were divided into 3 groups: control, H2O2 (with H2O2 concentration of 150 μM, cultured 12 hours), and H2O2+3-MA (10 mM of autophagy inhibitor 3-methyladenine pretreatment for 12 hours, then adding 150 μM H2O2 and cultured 12 hours). After 12 hours the cell autophagy-related protein LC3 (LC3-II/I ratio), Beclin-1, and p62 were tested by western blotting, cell vitality was tested by MTT, and apoptosis was tested by TUNEL assay and western blotting. RESULTS: Compared with the control group, the expression of Beclin-1 and LC3 (LC3-II/I ratio) increased, and the level of p62 decreased in the H2O2 group. Compared with the control group, the vitality of RPE-19 cells was decreased, and the apoptosis was increased in the H2O2 group (p<0.05). Compared with the H2O2 group, the expression of Beclin-1 and LC3 (LC3-II/I ratio) were decreased in the H2O2+3-MA group, while the expression of p62 was increased, RPE-19 cell vitality was increased, and cell apoptosis was decreased (p<0.05). CONCLUSION: H2O2 induced the activation of RPE-19 cell autophagy, the decrease of cell vitality, and the increase of cell apoptosis. Inhibition of autophagy can lead to the decrease of cell apoptosis. Autophagy may be involved in the apoptosis of RPE cells in the condition of oxidative stress. © Science Printers and Publishers Inc.
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