Fig 1: NOX4/EphA2 level was significantly elevated in pneumonia patients, especially in pneumonia patients using ventilator. A NOX4 concentration. B EphA2 concentration. *p < 0.05, and **p < 0.01, analyzed by two-way ANOVA with post-hoc testing and Bonferroni correction. Pt patients, MV mechanical ventilator
Fig 2: NOX4 is associated with EphA2 expression in VILI model. A Total cell counts and protein concentrations. B Identification of cells in BALF with representative visual fields. Cells were mainly macrophages (black arrow) with a few lymphocytes (blue arrow), and neutrophils (red arrow). C Histopathologic features of hematoxylin and eosin-stained mouse lung tissues and lung injury score, showing vascular engorgement, hyaline membrane formation, alveolar wall edema, and inflammatory cell infiltration (200 × magnification). D Western blot of EphA2 and NOX4, and densitometry of EphA2 and NOX4 in response to HTV and EphA2 KO. N = 3 in NVC group and n = 5 in HTV group. *p < 0.05, **p < 0.01, and ***p < 0.001, analyzed by two-way ANOVA with post-hoc testing and Bonferroni correction. BALF bronchoalveolar lavage fluid, WT wild-type, KO knockout, NVC normal ventilation control, HTV high tidal ventilation
Fig 3: NOX4 KO and NOX4 inhibitor attenuates VILI in mice. A Total cell counts and protein concentrations. B Cell differentiation in BALF with a representative visual field. Cells were mainly macrophages (black arrow) with a few lymphocytes (blue arrow) and neutrophils (red arrow). C Histopathologic features of hematoxylin and eosin-stained mouse lung tissues and lung injury score showing the extent of vascular engorgement, hyaline membrane formation, alveolar wall edema, and inflammatory cell infiltration (200 × magnification). D Relative levels of NOX4 mRNA determined by real-time PCR (n = 3, each group) and NOX4 immunohistochemistry (400 × magnification). E Inflammatory cytokine levels; IL-1ß, IL-6, and IL-8 levels in lung tissue lysates. N = 8 in each group. *p < 0.05, **p < 0.01, and ***p < 0.001, analyzed by two-way ANOVA with post-hoc testing and Bonferroni correction. BALF bronchoalveolar lavage fluid, NVC normal ventilation control, KO knockout, HTV high tidal ventilation
Fig 4: VILI is decreased in the NOX4 KO mouse and NOX4 inhibitor groups. Western blot of NOX4 (A), EphA2, and PI3K 110? (B). C Densitometry of EphA2, and PI3K 110?. *p < 0.05, **p < 0.01, and ***p < 0.001, analyzed by two-way ANOVA with post-hoc testing and Bonferroni correction. KO knockout, HTV high tidal ventilation
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