NADPH oxidases and reactive oxygen species at different stages of chronic hypoxia-induced pulmonary hypertension in newborn piglets

• Published in: American journal of physiology. Lung cellular and molecular physiology Vol. 297; no. 4; pp. L596 - L607
• Main Authors: Dennis, Kathleen E, Aschner, J. L, Milatovic, D, Schmidt, J. W, Aschner, M, Kaplowitz, M. R, Zhang, Y, Fike, Candice D
• Format: Journal Article
• Language: English
• Published: United States American Physiological Society 01.10.2009
• Subjects: Animals; Animals, Newborn; Antioxidants - pharmacology; Catalase - metabolism; F2-Isoprostanes - pharmacology; Hogs; Hypertension; Hypertension, Pulmonary - etiology; Hypertension, Pulmonary - metabolism; Hypoxia; Immunoblotting; Luminescence; Lungs; Manganese; NADPH Oxidase 1; NADPH Oxidases - metabolism; Organometallic Compounds - pharmacology; Oxidation; Oxygen; Phosphoproteins - metabolism; Pulmonary arteries; Pulmonary Artery - drug effects; Reactive Oxygen Species - metabolism; Superoxide Dismutase - metabolism; Superoxide Dismutase-1; Swine; Vascular Resistance - drug effects
• ISSN: 1040-0605; 1522-1504
• DOI: 10.1152/ajplung.90568.2008

1 Department of Pediatrics, ; 2 Vanderbilt Kennedy Center, ; 3 Center for Molecular Toxicology, and ; 4 Department of Pharmacology, Vanderbilt University Medical Center, Nashville, Tennessee Submitted 18 November 2008 ; accepted in final form 7 July 2009 Recently, we reported that reactive oxygen species (ROS) generated by NADPH oxidase (NOX) contribute to aberrant responses in pulmonary resistance arteries (PRAs) of piglets exposed to 3 days of hypoxia ( Am J Physiol Lung Cell Mol Physiol 295: L881–L888, 2008). An objective of the present study was to determine whether NOX-derived ROS also contribute to altered PRA responses at a more advanced stage of pulmonary hypertension, after 10 days of hypoxia. We further wished to advance knowledge about the specific NOX and antioxidant enzymes that are altered at early and later stages of pulmonary hypertension. Piglets were raised in room air (control) or hypoxia for 3 or 10 days. Using a cannulated artery technique, we found that treatments with agents that inhibit NOX (apocynin) or remove ROS [an SOD mimetic (M40403 ) + polyethylene glycol-catalase] diminished responses to ACh in PRAs from piglets exposed to 10 days of hypoxia. Western blot analysis showed an increase in expression of NOX1 and the membrane fraction of p67phox. Expression of NOX4, SOD2, and catalase were unchanged, whereas expression of SOD1 was reduced, in arteries from piglets raised in hypoxia for 3 or 10 days. Markers of oxidant stress, F 2 -isoprostanes, measured by gas chromatography-mass spectrometry, were increased in PRAs from piglets raised in hypoxia for 3 days, but not 10 days. We conclude that ROS derived from some, but not all, NOX family members, as well as alterations in the antioxidant enzyme SOD1, contribute to aberrant PRA responses at an early and a more progressive stage of chronic hypoxia-induced pulmonary hypertension in newborn piglets. superoxide dismutase enzymes; SOD1; SOD2; NOX4; NOX1; p67phox; catalase; F 2 -isoprostanes; M40403 Address for reprint requests and other correspondence: C. D. Fike, Dept. of Pediatrics, Vanderbilt Univ. Medical Center, 2215 B Garland Ave., 1125 MRB IV/Light Hall, Nashville, TN 37232-0656 (e-mail: Candice.fike{at}vanderbilt.edu ).