WWOX alleviates acute lung injury by suppressing macrophage glycolysis and mitochondrial dysfunction through HIF-1α lactylation

Background This study investigated whether WW domain-containing oxidoreductase (WWOX) confers protection against acute lung injury (ALI) by modulating hypoxia-inducible factor-1α (HIF-1α) lactylation, thereby influencing macrophage glycolysis and mitochondrial dysfunction. Methods ALI was modeled in vivo in mice and in vitro with lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. WWOX overexpression in vivo was achieved via adeno-associated virus (AAV), whereas stable WWOX-overexpressing and knockdown macrophage cell lines were generated through lentiviral and plasmid transduction. Multiple analytical approaches, including Western blotting, co-immunoprecipitation, immunofluorescence, liquid chromatography–mass spectrometry, Seahorse XF metabolic analysis, mitochondrial membrane potential assays, and histopathological analysis, were used to assess HIF-1α lactylation, glycolytic activity, mitochondrial function, and lung inflammatory injury. Results WWOX expression was markedly downregulated in the lung tissues of mice with ALI and LPS-stimulated RAW264.7 macrophages. WWOX overexpression attenuated HIF-1α transcriptional activity, leading to downregulation of key glycolytic enzymes (HK2, PKM2, and GLUT1), reduced lactate accumulation, and improved mitochondrial function by inhibiting DRP1 (dynamin-related protein 1) mediated fission and enhancing MFN2 (mitofusin 2) /OPA1 (optic Atrophy 1) dependent fusion. These changes ultimately alleviated pulmonary edema, inflammatory cytokine release, and histopathological damage. In contrast, WWOX knockdown led to increased glycolytic activation, aggravated mitochondrial dysfunction, and exacerbated lung inflammatory injury. Conclusion WWOX mitigates ALI by modulating HIF-1α lactylation, effectively suppressing pathological macrophage and mouse lung tissue glycolysis and excessive mitochondrial fission. This study delineates a novel metabolic–epigenetic regulatory axis implicated in the pathogenesis of ALI.