Autophagy: a new mechanism for regulating VEGF and PEDF expression in retinal pigment epithelium cells
Autophagy: a new mechanism for regulating VEGF and PEDF expression in retinal pigment epithelium cells作者机构：Department of Ophthalmology the First Affiliated Hospital of Xi'an Medical University Department of Ophthalmology Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University Department of Central Laboratory the First Affiliated Hospital of Xi'an Medical University Department of Ophthalmology the First Hospital of Yulin
出 版 物：《International Journal of Ophthalmology(English edition)》 (国际眼科杂志（英文版）)
年 卷 期：2019年第12卷第4期
页 面：557-562页
核心收录：
基 金：Supported by the National Natural Science Foundation of China(No.81500726) the Medical Research Project of Xi’an Science Technology Bureau [No.201805097YX5SF31(4)] the Health Research Foundation of Shaanxi Province(No.2018D074) the Outstanding Youth Talent Support Plan of Shaanxi Ordinary University
主 题：autophagy retinal pigment epithelium cells vascular endothelial growth factors pigment epithelium-derived factor angiogenesis
摘 要：AIM: To investigate the regulation of vascular endothelial growth factors(VEGF) and pigment epithelium-derived factor(PEDF) expression by autophagy in retinal pigment epithelium(RPE) cells on exposure to hypoxia. METHODS: ARPE-19, an RPE cell line, was treated as following: the control group was kept in a normoxic incubator; the hypoxia group was incubated in a hypoxic incubator with 1% O_2/5% CO_2/94% N_2 for 24 h; the hypoxia + 3-methyladenine(3-MA) group was pretreated with 10 mmol/L 3-MA for 1 h and then in the hypoxic incubator for 24 h; and the hypoxia + chloroquine(CQ) group was pretreated with 50 μmol/L CQ for 1 h and then in the hypoxic incubator for 24 h. The morphology and ultrastructure of the cells was observed by an inverted microscope or a transmission electronic microscope(TEM). Western blot was performed to assay the expression of autophagy-associated markers, including microtubule associated protein 1 light chain 3 B(LC3 B), Beclin-1, Atg5 and p62. The concentration of VEGF and PEDF in the culture supernatant was determined by ELISA, and the ratio of VEGF/PEDF was calculated. RESULTS: There were no obvious differences in cell morphology among different groups and autolysosomes could be observed in the cytoplasm in all groups. Compared to the control cells, the LC3 B-II/I ratio and levels of Beclin-1 and Atg5 were significantly increased and p62 level was significantly decreased in the hypoxia group. With the increase of VEGF and decrease of PEDF concentration, the VEGF/PEDF ratio was significantly increased in the hypoxia group compared to the control cells. The LC3 B-II/I ratio was significantly reduced by 3-MA treatment and increased by CQ treatment. The expressions of Beclin-1 and Atg5 were significantly reduced by 3-MA or CQ treatment, while expression of p62 was increased in the 3-MA or CQ treated cells. The concentration of VEGF was significantly decreased and PEDF increased, thereby the VEGF/PEDF ratio was decreased in the hypoxia + 3-MA gro
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