Overexpression of Annexin A1 Inhibits Pyroptosis and Improves Dry Eye Signs by Regulating the TRIM72/Nrf2/HO-1 Signaling Pathway.

This study aimed to investigate the therapeutic potential of annexin A1 (ANXA1) overexpression in improving the signs of dry eye disease (DED) and to elucidate the underlying molecular mechanism. A murine model of DED was established by topical application of 0.2% benzalkonium chloride (BAC), and human corneal epithelial (HCE-T) cells were exposed to 0.0005% BAC for in vitro experiments. ANXA1 was overexpressed using adenoviral vectors, and the effects on tear production, pyroptosis, and activation of the tripartite motif-containing protein 72 (TRIM72)/nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway were evaluated using Western blotting, Schirmer test, Terminal deoxynucleotidyl Transferase dUTP Nick End Labeling (TUNEL) assays, and fluorescent probe analyses. To further examine the role of TRIM72, its expression was silenced with specific small interfering RNA (siRNA), and the consequent impact on ANXA1-mediated therapeutic effects was assessed. ANXA1 expression was significantly reduced in both in vivo and in vitro DED models. Restoration of ANXA1 through overexpression significantly improved tear secretion and suppressed pyroptosis in the murine model. Similarly, in HCE-T cells, ANXA1 overexpression not only enhanced cellular proliferation but also significantly inhibited pyroptosis. Mechanistic investigations demonstrated that ANXA1 overexpression activated the TRIM72/Nrf2/HO-1 signaling pathway by increasing TRIM72, Nrf2, and HO-1 expression. Notably, silencing TRIM72 abolished the therapeutic effects of ANXA1 overexpression, thereby confirming that activation of this pathway is essential for mediating the protective effects of ANXA1 against DED. Overexpression of ANXA1 inhibits pyroptosis and improves dry eye signs by regulating the TRIM72/Nrf2/HO-1 axis.