Ophthalmology in China ›› 2023, Vol. 32 ›› Issue (4): 344-350.doi: 10.13281/j.cnki.issn.1004-4469.2023.04.014

Previous Articles     Next Articles

Hypoxia protects human lens epithelial cells from senescence induced by oxidative stress

Hou Fang1, Huang Qiming2, 3, Wan Xiuhua1   

  1. 1 Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University; Beijing Key Laboratory of Ophthalmology and Visual Science, Beijing 100730, China; 2 School of Life and Science, Nanchang University, Nangchang 330031, China; 3 Jiangxi Mayobio Technology Company Ltd., Nanchang 330006, China
  • Received:2022-12-05 Online:2023-07-25 Published:2023-07-25
  • Contact: Wan Xiuhua, Email: wanxh@ccmu.edu.cn
  • Supported by:
    The Training Plan for Academic and Technological Leaders of the Main Subject, Jiangxi Province (20204BCJ22035); Key Research & Development Project of Science and Technology Plan of Department of Science and Technology, Jiangxi Province (20192ACB70004, 20201BBG71007); General Project of Science and Technology Plan of Department of Science and Technology, Jiangxi Province (20192BBH80015); National Natural Science Foundation of China (82171037)

Abstract:  Objective To analyze the protection of hypoxia on the aging of human lens epithelial cells (hLECs) induced by 4-hydroxynonaenoic acid (4-HNE). Design Experimental research. Participants  Hypoxia and hLECs. Methods hLECs were treated with 4-HNE at different O2 concentrations and their proliferative activity were detected by CCK-8. The apoptosis levels, reactive oxygen species (ROS) and senescence of hLECs in hypoxic and normoxic condition were analyzed by Annexin-V/PI staining, fluorescent probe (DCFH-DA), β-galactosidase staining and BrdU combined assay respectively. Main Outcome Measures Proliferation activity, apoptosis, senescence and expression level of related proteins of hLECs. Results By compared with normal oxygen concentration, the cell viability of 1% hypoxia was significantly higher than that of 21% normoxia at 48 h, 72 h and 96 h, especially at 50 μM 4-HNE concentration at 96 h (4.75±0.45% and 2.17±0.52%, P=0.008). The survival rate of hLECs under hypoxia was significantly higher than that under normoxia (39.92±5.31% and 10.67±3.22% at 50 μM 4-HNE, P=0.003) detected by flow cytometry. The early and late apoptosis rates were significantly lower than those of normoxia (33.83±4.32% and 48.81±4.99% at 50 μM 4-HNE, P=0.015; 13.85±2.47% and 21.52±3.62% at 50 μM 4-HNE, P=0.007). The level of reactive oxygen species decreased significantly under hypoxia, which was significantly lower than that of normal oxygen (63.44±8.73% and 28.85±5.77% at 50 μM 4-HNE, P=0.009). Aging related β-galactosidase positive rate was significantly lower than those of normoxia (35.31±4.13% and 76.4±4.21% at 50 μM 4-HNE, P=0.004) and the positive of BrdU incorporation significantly higher than those of normoxia (6.55±1.74% and 3.20±0.93% at 50 μM 4-HNE, P=0.021). Western blot results showed that 4-HNE treatment could activate the expression of P53 and P16, the main regulators of aging. However, compared with the hypoxia group, the relative expression of P16 and P53 protein in the normoxia group was significantly higher than that in the hypoxia group (P=0.031, 0.005). Conclusion Hypoxia can promote the proliferation of hLECs, inhibit the apoptosis. ROS and aging level of hLECs caused by oxidative stress, and protect hLECs. (Ophthalmol CHN, 2023, 32: 344-350)

Key words: hypoxia, lens epithelial cells, 4-hydroxynonenoic acid, aging, oxidative stress