Study of the mechanism of methyltransferase 3 regulation of ferroptosis in allergic rhinitis and promotion of allergic rhinitis in an m6A-dependent mechanism

Objective This study aimed to investigate the regulation of allergic rhinitis (AR) by methyltransferase 3 through an m6A-dependent mechanism, providing a theoretical foundation for its treatment. Methods An in vitro experiment was conducted in which HNEpC cells were stimulated with IL-13 (50 ng/mL) to create an AR cell model. After establishing the AR cell model, the cells were treated with DAA (m6A inhibitor) and separated into three groups: Control group, shNC group and shMETTL3 group.m6A-RIP assessed the m6A modification level of PTBP1 mRNA, while RIP was used to analyze the interaction between METTL3 and PTBP1 mRNA. The mice were grouped as follows: AR, AR + shNC, AR + shMETTL3, AR + shMETTL3 + OE-PTBP1, and AR + shMETTL3 + OE-TXNIP Serum concentrations of INF-γ, IL-1β, IL-18, TGF-β, IL-4, IL-10, IgE, IgG2a, and IgG1, along with oxidative stress markers (ROS, MDA, SOD, GSH), were measured using ELISA. Additionally, ferroptosis-related proteins (GPX4, Nrf2, MnSOD, and ASCL4) in the nasal mucosa were analyzed using western blotting. Results In vitro cell experiments demonstrated that shMETTL3 decreased both m6A modification and expression levels of PTBP1.The model group mice exhibited higher behavioral scores than the normal group, whereas the shMETTL3 group showed significantly lower scores. Animal experiment results indicated that, relative to the sham group, the AR group exhibited significant increases in serum levels of INF-γ, IL-18, IL-1β, IL-4, IL-10, IgE, IgG2a, IgG1, MDA, and ROS, while TGF-β, SOD, and GSH levels were significantly reduced. In the shMETTL3 group, serum levels of INF-γ, IL-18, IL-1β, IL-4, IL-10, IgE, IgG2a, IgG1, SOD, and GSH were significantly reduced, while TGF-β, SOD, and GSH levels were significantly elevated than those in the AR group. Additionally, GPX4 and MnSOD protein expression in the nasal mucosa increased significantly, whereas Nrf2, ASCL4, METTL3, PTBP1, and TXNIP protein expression decreased significantly. In the AR + shMETTL3 + OE-TXNIP group, serum levels of INF-γ, IL-18, IL-1β, IL-4, IL-10, IgE, IgG2a, IgG1, MDA, and ROS were significantly elevated, whereas TGF-β, SOD, and GSH levels were notably reduced compared to the shMETTL3 group. Conclusion Methyltransferase 3 potentially modulates ferroptosis and oxidative stress linked to AR through an m6A-dependent mechanism, thereby alleviating symptoms in AR mice.