1. Nox1 as a potential therapeutic target to inhibit matricellular-mediated endothelial senescence.
Nox1 as a potential therapeutic target to inhibit matricellular-mediated endothelial senescence. Data for Nox1OE are shown in blue; data for Nox1−/− samples are shown in orange; data for cell experiments are shown in red; data in gray are the corresponding controls. (A and B) Effect of human Nox1OE on SA-β-Gal staining and O2•− production. Graphical data are means ± SEM (n = 3 biological replicates per treatment). Scale bar, 40 μm. *P < 0.05 for Nox1 plasmid compared to empty plasmid control by Student’s t test. (C) NADPH-driven O2•− production, as measured by cytochrome c reduction assay in middle-aged wild-type and Nox1−/− mouse lungs. Left: Representative cytochrome c kinetic curve. Right: Quantification of O2·− production. Bar graphs are means ± SEM (n = 6 mice per group). *P < 0.05 compared to wild-type by Student’s t test. (D) mRNA expression of p53 and p21cip in wild-type and Nox1−/− mouse lungs. Data are the means ± SEM (n = 3 mice per group); *P < 0.05 compared to wild type by Student’s t test. (E to G) Effect of Nox1 inhibition (using NoxA1ds) on TSP1-induced cell cycle arrest in HPAECs, as measured by MTT assay (E), trypan blue exclusion assay (F), and cell cycle profile analysis (G). Graphical data are means ± SEM (n = 3 to 4 biological replicates per treatment). *P < 0.05 for TSP1 challenge compared to scrambled vehicle control (SCRAMB) by one-way ANOVA. (H) Activation of p53 (red), as assessed by nuclear localization (DAPI-labeled, blue) and p21cip immunofluorescence (green) in HPAECs challenged with TSP1 in the presence or absence of the Nox1 inhibitor NoxA1ds. Graphical data are means ± SEM (n = 3 biological replicates per treatment, 12 to 20 images per group). Scale bar, 50 μm. *P < 0.05 for TSP1 challenge compared to scrambled vehicle control by one-way ANOVA.
Daniel N. Meijles et al., Sci. Signal. 2017;10:eaaj1784
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