Fig. 7
NOX2 deletion or pharmacological inhibition dampens pathologically high intraocular pressure injury-mediated reactive glial cell proliferation and neuroinflammation and identifies inflammatory and immune-related protein pathways and functional enrichment in experimental glaucoma. (A) Representative images of GFAP stained retinal cryosections from WT and Nox2−/− mice after Pressure 60 mmHg 24 h. Scale bar, 50 μm. (B) Analysis of GFAP fluorescence mean intensity/µm2 (n = 6 in each group). (C) Representative images of Iba1 stained retinal flat-mounts and retinal cryosections from WT and Nox2−/− mice after Pressure 60 mmHg 24 h. Scale bar, 50 μm. (D) Analysis of the number of Iba1 + cells/mm2, Iba1 + percent area, the mean dendrite length (µm), the mean number of branches (n), and the number of Iba1 positive cells per section (n = 6 in each group). (E) ELISA assay was performed to determine the retinal protein of TNF-α and IL-6 with gp91ds-tat treatment (n = 12 in each group). (F) Messenger RNA expression of genes coding for proinflammatory cytokines (Tnf-α and Il-1β) in different groups. Data are presented as the fold-change versus Veh-control (n = 6 in each group). (G) Messenger RNA expression of hypoxia genes (Vegf-a), antioxidant genes (Ho-1, Gpx1, Sod2), and nitric oxide synthases (eNos, iNos, and nNos) in different groups. Data are presented as the fold-change in retinas after H-IOP and with gp91ds-tat treatment versus control-vehicle (n = 6 in each group). Data are shown as mean ± SEM (one-way ANOVA with Tukey’s multiple comparisons test, *p < 0.05, **p < 0.01,***p < 0.001, ****p < 0.0001)