Paired Box Gene 6 Regulates Heme Oxygenase-1 Expression and Mitigates Hydrogen Peroxide-Induced Oxidative Stress in Lens Epithelial Cells

• Published in: Current eye research Vol. 47; no. 11; pp. 1516 - 1524
• Main Authors: Zheng, Yuxing, Yang, Xiaoxi, Wu, Shuduan, Yi, Guoguo, Huang, Xiaobo, Feng, Zhizhen, Qu, Lei, Liu, Linyu, Li, Qianyun, Xia, Zhaoxia
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 02.11.2022
• Subjects: Age-related cataract; Animals; Annexin A5 - metabolism; Antioxidants - metabolism; Apoptosis; Capsules - metabolism; Epithelial Cells - metabolism; Fluoresceins - metabolism; heme oxygenase-1; Heme Oxygenase-1 - genetics; Heme Oxygenase-1 - metabolism; Hydrogen Peroxide - metabolism; Isothiocyanates; lens epithelial cells (SRA01/04 HLE-B3); Lens, Crystalline - metabolism; Membrane Proteins; Mice; Oxidative Stress; paired box gene 6; PAX6 Transcription Factor; Propidium - metabolism; Reactive Oxygen Species - metabolism; RNA, Small Interfering - genetics; Superoxide Dismutase - genetics; Superoxide Dismutase - metabolism; heme oxygenase-1; oxidative stress; lens epithelial cells (SRA01/04 HLE-B3); Age-related cataract; paired box gene 6
• ISSN: 0271-3683; 1460-2202
• DOI: 10.1080/02713683.2022.2110266

This study aimed to investigate the regulation of heme oxygenase-1 (HO-1) by paired box gene 6 (Pax6) and their roles in hydrogen peroxide (H 2 O 2 )-induced oxidative stress and apoptosis in lens epithelial cells (LECs) (SRA01/04, HLE-B3). Lens anterior capsule membranes of mice of different ages were obtained to compare differences in the expression of Pax6 and HO-1 using Western blotting. Pax6-overexpressing plasmid and small interfering RNA were designed to overexpress and silence Pax6, respectively. Cobalt protoporphyrin (CoPP) was used to promote the expression of HO-1. Oxidative damage in LECs was induced by treatment with H 2 O 2 (400 µM) for 24 h. Cell viability was measured using the Cell Counting Kit-8 assay. Intracellular reactive oxygen species (ROS) were detected using flow cytometry and immunofluorescence. Superoxide dismutase (SOD) level was measured using SOD Assay Kit and apoptotic cells were quantified using annexin V-fluorescein isothiocyanate/propidium iodide staining. Pax6 and HO-1 expression levels showed an age-dependent decrease in LECs of mouse. Overexpressing Pax6 upregulated HO-1 expression level. Silencing Pax6 downregulated the HO-1 expression level, resulting in increased generation of ROS, reduced SOD activity, decreased cell viability, and increased apoptotic cells of LECs under H 2 O 2 -induced oxidative stress. Overexpressing Pax6 and CoPP both mitigates H 2 O 2 -induced oxidative stress by increasing the expression of HO-1 of LECs. Pax6 and HO-1 expression levels showed an age-dependent decrease in LECs in mouse anterior capsules. Pax6 could regulate the expression of HO-1 in LECs. The decrease of Pax6 weakened the antioxidant ability of LECs under H 2 O 2 -induced oxidative stress by downregulating HO-1, which may be a potential mechanism for the formation of age-related cataract.