Warifteine Alleviates Acute Lung Injury and Inhibits NETosis by Decreasing Reactive Oxygen Species–induced Gasdermin D Oligomerization

Acute lung injury (ALI) is characterized by exaggerated inflammatory reactions and high clinical mortality rates, but targeted therapeutic interventions are lacking. Warifteine, which is a traditional remedy known for its antiinflammatory properties, has been identified, but its potential effects on ALI remain unexplored. In this study, a murine model of ALI was established by injecting LPS into wild-type (WT) or neutrophil-specific <i>Gsdmd</i> (Gasdermin D)-deficient mice. Pulmonary function was evaluated, and lung samples were collected for immunofluorescence staining and RNA sequencing analysis. Additionally, live imaging of the lungs as well as histological, biochemical, and molecular investigations were performed to assess the progression of LPS-induced ALI in mice. Mouse bone marrow-derived neutrophils were isolated and cultured to investigate the effects of warifteine <i>in vitro</i>. Our findings indicate that warifteine effectively mitigates LPS-induced lung pathology and dysfunction in mice. Mechanistic studies revealed that warifteine protects against ALI by inhibiting neutrophil extracellular trap formation and the resulting cytokine storm. Subsequent research demonstrated that warifteine influences neutrophil extracellular trap formation by inhibiting GSDMD oligomerization through the regulation of reactive oxygen species production by neutrophils. Collectively, these findings reveal a novel role for warifteine in the protection against ALI and suggest that modulating GSDMD oligomerization may be an innovative therapeutic strategy.