The Effect of Heparinase Inhibitor PI-88 on Human Retinal Endothelial Cells Autophagy under Hypoxia
- VernacularTitle:缺氧条件下乙酰肝素酶抑制剂PI-88 对人视网膜血管内皮细胞自噬的影响
- Author:
Yu-xin YUAN
1
;
Shu-hua LI
2
;
Meng XUAN
1
;
Kun-yi SU
1
;
Lin LV
1
;
Jie HU
1
Author Information
1. State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China
2. Guangdong Province Traditional Chinese Medical Hospital, The Second Affiliated Hospital of Traditional Chinese Medicine University Of Guangzhou, Guangzhou 510120, China
- Publication Type:Journal Article
- Keywords:
heparinase;
PI-88;
human retinal endothelial cells;
autophagy;
hypoxia
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2020;41(3):397-402
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】 To explore the potential role of heparinase inhibitor PI-88 in autophagy of human retinal endothelial cells under hypoxia. 【Methods】 The human retinal endothelial cells were cultured in vitro and divided into normal culture group, hypoxia group(CoCl2), and hypoxia plus heparinase inhibitor(CoCl2+PI-88) group. Immunofluorescence was used to observe the expression of LC3. Western blot was adopted to assess the LC3-Ⅰ and LC3-Ⅱ expression. Green fluorescent protein(GFP) -LC3 adenoviruses were transfected into living human retinal endothelial cells to dynamically visualize the autophagy level. 【Results】 LC3 exposed to hypoxia were upregulated, the ratio of LC3-II/LC3-I in HREC were upregulated(grey value, hypoxia group vs. normal culture group, 4.69±0.5 vs. 2.14±0.33, P = 0.001 < 0.05). The ratio of LC3-II/LC3-I in HREC were inhibited in response to heparinase inhibitor PI-88(grey value, hypoxia plus heparinase inhibitor group vs hypoxia group, 3.51±0.20 vs. 4.69±0.54, P = 0.027 < 0.05). Autophagosome marked by LC3-GFP in HREC exposed to heparinase inhibitor(PI-88) were decreasing compared to cells only exposed to hypoxia. 【Conclusion】 Inhibiting heparinase could downregulate the level of autophagosome in human retinal endothelial cells under hypoxia.
