Hydrogen sulfide protects retinal pigment epithelium cells against ferroptosis through the AMPK- and p62-dependent non-canonical NRF2-KEAP1 pathway

Abstract

Oxidative stress-induced ferroptosis of retinal pigment epithelium (RPE) cells contributes to retinal degenerative diseases. The antioxidant molecule hydrogen sulfide (H₂S) regulates oxidative stress response, but its effect on the ferroptosis of RPE cells is unclear. In this study, sodium hydrosulfide (NaHS) was used as an exogenous H₂S donor to intervene tert-butyl hydroperoxide (t-BHP)-induced ferroptosis of APRE-19 cells. We found that NaHS pretreatment attenuates t-BHP-induced oxidative stress and ferroptosis. Analysis of mRNA-sequencing coupled with FerrDb database identified nuclear factor erythroid-2-related factor 2 (NRF2) as a primary target for the cytoprotective role of H₂S. NRF2 inhibitor ML385 reverses the effects of H₂S on ferroptosis. Biochemical analysis revealed that H₂S stabilizes NRF2. H₂S decreases the interaction between NRF2 and KEAP1, but enhances the interaction between KEAP1 and p62. These results suggest that H₂S activates the non-canonical NRF2-KEAP1 pathway. Further study demonstrated that H₂S stimulates AMPK to interact and phosphorylate p62. Additionally, inhibiting AMPK or knocking down p62 blocks the effects of H₂S. We speculate that targeting the non-canonical NRF2-KEAP1 pathway by H₂S-based drug may benefit the treatment of retinal degenerative diseases.

Keywords: Ferroptosis; Hydrogen sulfide; NRF2; Oxidative stress; RPE.