Pentaerythritol Tetranitrate In Vivo Treatment Improves Oxidative Stress and Vascular Dysfunction by Suppression of Endothelin-1 Signaling in Monocrotaline-Induced Pulmonary Hypertension

Figure 5

Effects of PETN therapy on prooxidative protein activity (Nox, XO), oxidative burst in whole blood, and antioxidant HO-1 mRNA expression in pulmonary hypertension (MCT40, 4 weeks). (a) Cardiac NADPH oxidase (Nox) activity was measured by the chemiluminescence probe lucigenin (5 µM) in the presence of NADPH (200 µM). (b) Leukocyte-derived oxidative burst in whole blood was examined by the chemiluminescence probe L-012 (100 µM) upon stimulation with PDBu (10 µM). (c) Xanthine oxidase (XO) activity was assessed by a photometric assay using cytochrome c (change in absorbance: ΔA 550 nm). qRT-PCR was used to determine mRNA expression levels of the antioxidant enzyme (d) heme oxygenase-1 (HO-1) and (e) vascular adhesion molecule-1 (VCAM-1) in lung tissue. (f) NADPH oxidase 2 (Nox2) protein expression was determined by Western blot analysis. The data are mean ± SEM from 3–6 animals/group. versus control; versus MCT40.

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