title: Selective inactivation of NADPH oxidase 2 causes regression of vascularization and the size and stability of atherosclerotic plaques.
abstract: BACKGROUND
  A variety of NADPH oxidase (Nox) isoforms including Noxs 1, 2, 4 and 5 catalyze the formation of reactive oxygen species (ROS) in the vascular wall. The Nox2 isoform complex has arguably received the greatest attention in the progression of atherogenesis in animal models. Thus, in the current study we postulated that specific Nox2 oxidase inhibition could reverse or attenuate atherosclerosis in mice fed a high-fat diet.
  
  
  METHODS
  We evaluated the effect of isoform-selective Nox2 assembly inhibitor on the progression and vascularization of atheromatous plaques. Apolipoprotein E-deficient mice (ApoE-/-) were fed a high fat diet for two months and treated over 15 days with Nox2ds-tat or control sequence (scrambled); 10 mg/kg/day, i.p. Mice were sacrificed and superoxide production in arterial tissue was detected by cytochrome C reduction assay and dihydroethidium staining. Plaque development was evaluated and the angiogenic markers VEGF, HIF1-α and visfatin were quantified by real time qRT-PCR. MMP-9 protein release and gelatinolytic activity was determined as a marker for vascularization.
  
  
  RESULTS
  Nox2ds-tat inhibited Nox-derived superoxide determined by cytochrome C in carotid arteries (2.3 ± 0.1 vs 1.7 ± 0.1 O2(•-) nmol/min*mg protein; P < 0.01) and caused a significant regression in atherosclerotic plaques in aorta (66 ± 6 μm(2) vs 37 ± 1 μm(2); scrmb vs. Nox2ds-tat; P < 0.001). Increased VEGF, HIF-1α, MMP-9 and visfatin expression in arterial tissue in response to high-fat diet were significantly attenuated by Nox2ds-tat which in turn impaired both MMP-9 protein expression and activity.
  
  
  CONCLUSIONS
  Given these results, it is quite evident that selective Nox inhibitors can reverse vascular pathology arising with atherosclerosis.
authors: Vascular Biology Lab, Institute of Medicine and Experimental Biology of Cuyo (IMBECU) CONICET, School of Medical Sciences, National University of Cuyo, Mendoza, Argentina.
authors: I M
authors: IM
authors: Quesada
authors: Vascular Biology Lab, Institute of Medicine and Experimental Biology of Cuyo (IMBECU) CONICET, School of Medical Sciences, National University of Cuyo, Mendoza, Argentina.
authors: A
authors: A
authors: Lucero
authors: Cellular and Molecular Biology Lab, Institute of Histology and Embryology (IHEM) CONICET, Mendoza, Argentina.
authors: C
authors: C
authors: Amaya
authors: Department of Pharmacology & Chemical Biology and Vascular Medicine Institute, 200 Lothrop Street, University of Pittsburgh, Pittsburgh, PA 15261, USA.
authors: D N
authors: DN
authors: Meijles
authors: Department of Pharmacology & Chemical Biology and Vascular Medicine Institute, 200 Lothrop Street, University of Pittsburgh, Pittsburgh, PA 15261, USA.
authors: M E
authors: ME
authors: Cifuentes
authors: Department of Pharmacology & Chemical Biology and Vascular Medicine Institute, 200 Lothrop Street, University of Pittsburgh, Pittsburgh, PA 15261, USA.
authors: P J
authors: PJ
authors: Pagano
authors: Vascular Biology Lab, Institute of Medicine and Experimental Biology of Cuyo (IMBECU) CONICET, School of Medical Sciences, National University of Cuyo, Mendoza, Argentina. Electronic address: ccastro@fcm.uncu.edu.ar.
authors: C
authors: C
authors: Castro
mesh_terms: D000818:Animals; D001011:Aorta / Q000201:enzymology / Q000473:pathology; D001057:Apolipoproteins E / Q000235:genetics; D002339:Carotid Arteries / Q000473:pathology; D045304:Cytochromes c / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D059305:Diet, High-Fat; D018450:Disease Progression; D004791:Enzyme Inhibitors / Q000737:chemistry; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009389:Neovascularization, Pathologic; D054409:Nicotinamide Phosphoribosyltransferase / Q000378:metabolism; D016877:Oxidants / Q000737:chemistry; D018384:Oxidative Stress; D058226:Plaque, Atherosclerotic / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control* / Q000628:therapy*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012044:Regression Analysis; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D001057:Apolipoproteins E; D016207:Cytokines; D004791:Enzyme Inhibitors; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008562:Membrane Glycoproteins; D016877:Oxidants; D017382:Reactive Oxygen Species; D042461:Vascular Endothelial Growth Factor A; C467480:vascular endothelial growth factor A, mouse; D045304:Cytochromes c; C488842:Cybb protein, mouse; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D054409:Nicotinamide Phosphoribosyltransferase; C495867:nicotinamide phosphoribosyltransferase, mouse; D020780:Matrix Metalloproteinase 9; C501830:Mmp9 protein, mouse
keywords: Oxidative stress; Vascular Nox; Vulnerable plaque