Fig 2: Increased oxidative stress in peri-N from patients with CPA.(a) Western blot analysis of NOX4, Nrf2, HO-1 and ß-actin (loading control) protein levels in perirenal adipose tissue; NT peri-N, n = 6; CPA peri-N, n = 6. (b) Western blot analysis of phospho-NF-?B p65 and NF-?B p65 nuclear protein levels in perirenal adipose tissue. H3 was served as a protein loading control. (c, d) Expression changes in reactive oxygen species genes in sub-Q and peri-N from patients with CPA (n = 8) and normotension patients (n = 10). *P < 0.05 vs normotension subjects.

Fig 3: AIP1 expression is markedly reduced after alkali burn injury. A Representative slit-lamp images of the area of corneal neovascularization at different times after alkali burn injury (magnification: × 40). B Corneal opacity, neovessel size, and vessel size scores increased gradually from the third day of alkali burn injury and continued thereafter (N = nine). C Immunofluorescence staining revealed an obvious decrease in the expression of AIP1 and a concomitant increase in the expression of VEGFa after alkali burn injury (scale bar: 50 µm, magnification: × 400). D Western blot analysis demonstrating significantly upregulated expression of NOX4, NLRP3, ASC, clv-casp-1 and clv-IL-1ß and downregulated expression of AIP1 and NLRP6 in the cornea after alkali burn injury (N = three). The error bars represent the means ± SD, and comparisons were made using one-way ANOVA. clv-casp1, cleaved caspase-1; clv-IL-1ß, cleaved-IL-1ß. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001

Fig 4: Schematic diagram of the proposed mechanism. The protective effect of the NOX4 inhibitor and AIP1 on corneal neovascularization after alkali burn injury is associated with a reduction in ROS production and alleviation of the imbalance in NLRP3 activation and NLRP6 suppression

Fig 5: AIP1 depletion increases neovascularization, ROS production, and NOX4 expression and exacerbates the imbalance in NLRP3/NLRP6. A Representative slit-lamp images showing that AIP1 KO notably increased neovascularization compared with that in C57BL/6 mice (magnification: × 40). B Corneal whole-mount staining showing that AIP1 KO mice exhibited notably increased neovascularization compared with C57BL/6 mice (scale bar: 1 mm). C The corneal opacity, neovessel size, and vessel size scores increased significantly in AIP1-KO mice compared with those in C57BL/6 mice after alkali burn injury (N = nine). D The DCFDA ROS assay revealed that AIP1 KO notably increased ROS production after alkali burn injury compared with that in the control group (scale bar: 100 µm). E RT–qPCR analysis of AIP1 gene knockout efficiency. Deletion of AIP1 significantly increased the reduction in NLRP6 and elevation in NOX4, NLRP3 and VEGFa induced by alkali burn injury (N = three). F Western blot analysis showed that AIP1 deletion significantly increased the reduction in NLRP6 and elevation in NOX4 and NLRP3 induced by alkali burn injury (N = three). G Immunofluorescence staining showing that deletion of AIP1 significantly increased the elevation in VEGFa induced by alkali burn injury (scale bar: 50 µm; magnification: × 400). Error bars represent the mean ± SD, and comparisons were performed using one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001