RNF8 Relieves asthma-induced DNA Damage and Ferroptosis in Airway Epithelial Cells Through the AKT-NRF2 Axis

Asthma is a common chronic respiratory disease, and airway epithelial cell injury is closely related to the progression of asthma. Ferroptosis is a new type of cell death that has been proven to be involved in the pathogenesis of asthma. Notably, RNF8 has been confirmed to be critical for maintaining epithelial barrier integrity, but its role in asthma airway epithelial cell injury and ferroptosis remains unclear. Mice and 16HBE cells were treated with house dust mite (HDM) extract to establish in vivo and in vitro asthma models. HE staining and Masson staining were used to evaluate pathological lesions in mouse lung tissues, a CCK-8 assay was used to assess cell viability, and an alkaline comet assay was performed to evaluate DNA damage. The expression of key proteins was detected via Western blotting, immunohistochemistry, and immunofluorescence. Our research revealed that HDM extract treatment led to DNA damage and ferroptosis in both 16HBE cells and mouse lung tissue. Additionally, HDM extract treatment decreased RNF8 expression in 16HBE cells and mouse lung tissue, whereas RNF8 overexpression reduced the levels of γH2AX, 8-OHdG, Fe²⁺, MDA, and ROS in HDM extract-treated 16HBE cells and mouse lung tissue; increased SOD levels; increased GPX4 and SLC7A11 expression; and decreased ACSL4 expression. Furthermore, RNF8 overexpression decreased the p-PI3K/PI3K ratio and the p-AKT/AKT ratio but increased NRF2 expression in 16HBE cells treated with HDM extract. Research into the underlying mechanisms revealed that RNF8 may increase NRF2 expression by blocking the PI3K/AKT signaling pathway, thus alleviating DNA damage and inhibiting ferroptosis in airway epithelial cells, in turn slowing asthma progression. Our research revealed that RNF8 may slow the progression of asthma, providing a potential target for the treatment of asthma.