NOX1/4 drives hepatic iron and lipid dysregulation, redox imbalance, and inflammation in ethanol-fed mice.

Alcoholic liver disease (ALD) is pathologically characterized by iron overload, oxidative stress, metabolic dysregulation, and inflammation. However, the central regulatory mechanisms orchestrating these processes remain elusive. This study aimed to explore the role of NADPH oxidases 1 and 4 (NOX1/4) in ethanol-induced liver injury and to evaluate the therapeutic potential of the NOX1/4 inhibitor, Setanaxib. Mice were randomly divided into three groups: pair-fed control, ethanol-fed, and ethanol-fed + Setanaxib treatment. On day 16, the mice were euthanized, and liver tissue and blood samples were collected for comprehensive analyses to explore the involvement and potential mechanisms of NOX1/4 in ethanol-induced liver injury. Hepatic NOX1/4 expression was significantly upregulated in ethanol-fed mice, implicating their role in ALD pathogenesis. Setanaxib treatment markedly attenuated multiple facets of this injury, including liver injury, iron overload, hepatic steatosis, and oxidative stress. Mechanistically, Setanaxib may restore iron homeostasis by suppressing hepcidin/FPN1 signaling and downregulating TFR1 expression, activate antioxidant defenses via the Nrf2/HO-1/SLC7A11/GPX4 axis, and mitigate inflammation by reducing macrophage infiltration and pro-inflammatory cytokine release. Our findings suggest that NOX1/4 may play a central role in coordinating the interconnected pathogenic processes that drive ALD progression, including iron overload, lipid dysregulation, oxidative stress, and inflammation. Our findings establish Setanaxib, a novel NOX1/4 inhibitor, as a promising multi-target therapeutic strategy for ALD.