Abstract

Sepsis is a life-threatening syndrome driven by dysregulated thromboinflammation. Effective therapies are currently lacking due to the clinical limitations and bleeding risks associated with conventional anticoagulants. In this study, we demonstrate that Oroxylin A, a natural flavonoid, mitigates sepsis-associated coagulopathy by targeting arachidonate 12-lipoxygenase (ALOX12)-mediated lipid peroxidation. Using murine models of cecal ligation and puncture (CLP) and bacterial sepsis, we found that Oroxylin A significantly improved survival, restored platelet counts, and normalized coagulation parameters, including PT, APTT, and D-dimer levels. Furthermore, Oroxylin A reduced fibrin deposition and plasma tissue factor (F3) levels. Mechanistically, Oroxylin A promoted the ubiquitin-proteasome-dependent degradation of ALOX12, thereby suppressing lipid peroxidation markers (4-HNE and MDA) and the downstream interferon-beta (IFN-β)-driven transcription of F3. Additionally, Oroxylin A inhibited gasdermin D (GSDMD)-dependent pyroptosis, a key pathway for F3 release. Genetic ablation of ALOX12 abolished the therapeutic effects of Oroxylin A, confirming its target specificity. Molecular docking revealed that Oroxylin A binds to Asp632 within the catalytic domain of ALOX12, destabilizing the enzyme. These findings establish ALOX12 as a critical mediator of sepsis-associated coagulation and highlight the dual role of Oroxylin A in blocking pathological F3 expression and pyroptosis. This study provides a novel strategy for sepsis management by targeting lipid peroxidation upstream of thromboinflammatory cascades, offering improved safety over traditional anticoagulants.