Extracellular peroxiredoxin 6 released from alveolar epithelial cells as a DAMP drives macrophage activation and inflammatory exacerbation in acute lung injury.

Acute respiratory distress syndrome (ARDS) is featured with acute lung inflammatory injury. Our prospective study found that higher levels of peroxiredoxin 6(PRDX6) were detected in bronchoalveolar lavage (BAL) fluid from ARDS patients. Elevated PRDX6 was also correlated with monocytic activation and poor prognosis in ARDS patients. To investigate the origin of extracellular PRDX6, we conducted in vitro and in vivo experiments, demonstrating that PRDX6 can be actively released from alveolar epithelial cells under stress conditions. Our study demonstrated that it could be released from injured lung epithelial cells into the bronchoalveolar interstitial space in mice with acute lung injury and in vitro experiments. Moreover, exogenous PRDX6 was shown to activate the TLR4/NF-κB signalling pathway and induce M1 polarization of macrophages. Notably, the inflammatory effects of PRDX6 were mitigated by specific inhibition of the TLR4 (Toll-like receptor 4)-MD2 (Myeloid differentiation factor 2) complex. Using molecular docking simulations and in vitro binding assays, we confirmed a direct interaction between PRDX6 and MD2, further supporting its role as a damage-associated molecular patterns (DAMP) in ARDS. Our findings suggest that extracellular PRDX6 in bronchoalveolar lavage fluid could be a new DAMP factor in ALI, providing new insights into the pathogenesis of secondary hit in ALI/ARDS and highlighting PRDX6 as a potential therapeutic target for mitigating lung inflammation.