Vascular oxidant stress early after balloon injury: evidence for increased NAD(P)H oxidoreductase activity |
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| Authors: | Souza H P Souza L C Anastacio V M Pereira A C Junqueira M L Krieger J E da Luz P L Augusto O Laurindo F R |
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| Affiliation: | 1. Emergency Medicine Research Laboratory, University of São Paulo, São Paulo, Brazil;2. Heart Institute, (InCor), School of Medicine, University of São Paulo, São Paulo, Brazil;3. Instituto de Quı́mica, University of São Paulo, São Paulo, Brazil;4. Federal University of Florianópolis, Santa Catarina, Brazil;1. Molecular Physiology, Molecular Medicine Research Group, School of Medicine, Western Sydney University, Campbelltown Campus, NSW, 2560, Australia;2. Departments of Health Sciences and Biological Sciences, Faculties of Applied Health Sciences and Mathematics & Science, Brock University, St. Catharines, ON, L2S 3A1, Canada;3. Department of Chemical Engineering, Stanford University, Stanford, California 94305;5. Department of Chemistry, Stanford University, Stanford, California 94305;4. School of Medicine, Stanford University, Stanford, California 94305;6. Stanford ChEM-H, Stanford University, Stanford, California 94305;1. Department of Thoracic Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, PR China;2. Department of Pathogen Biology, Medical College, Nantong University, Nantong, Jiangsu 226001, PR China;3. Department of Radiotherapy, Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, PR China;4. Department of Thoracic Surgery, Rugao People’s Hospital, Rugao, Jiangsu 226500, PR China;5. Department of Vasculocardiology, Affiliated Hospital of Nantong University, Nantong, Jiangsu 226001, PR China;6. Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Medical College, Nantong University, Nantong, Jiangsu 226001, PR China;1. Department of Vascular Surgery, Centre Hospitalier Universitaire Vaudois (CHUV), Lausanne, Switzerland;2. Department of Surgery, Brigham and Women''s Hospital, Harvard Medical School, Boston, MA, USA;3. Department of Genetics and Complex Diseases, Harvard School of Public Health, Boston, MA, USA;4. Department of Vascular Surgery, Hôpital Pellegrin, Bordeaux, France;5. Department of Angiology, Centre Hospitalier Universitaire Vaudois (CHUV), Lausanne, Switzerland;1. Hematology Department, University Hospital of Salamanca, Biomedical Research Institute of Salamanca, Salamanca, Spain;2. Cancer Research Center, University of Salamanca, Salamanca, Spain;3. Bellvitge Biomedical Research Institute, Barcelona, Spain;4. Hematology Department, General Hospital of Segovia, Segovia, Spain;5. University Clinic of Navarra, Center of Applied Medical Research, Navarra Institute for Health Research, Pamplona, Spain |
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| Abstract: | Available evidence for oxidative stress after angioplasty is indirect or ambiguous. We sought to characterize the pattern, time course, and possible sources of free radical generation early after arterial balloon injury. Ex vivo injury performed in arterial rings in buffer with lucigenin yielded a massive oxygen-dependent peak of luminescence that decayed exponentially and was proportional to the degree of injury. Signals for injured vs. control arteries were 207. 1 +/- 17.9 (n = 13) vs 4.1 +/- 0.7 (n = 22) cpm x 10(3)/mg/min (p <. 001). Data obtained with 0.25 mmol/l lucigenin were validated with 0. 005-0.05 mmol/l lucigenin or the novel superoxide-sensitive probe coelenterazine (5 micromol/l). Gentle removal of endothelium prior to injury scarcely affected the amount of luminescence. Lucigenin signals were amplified 5- to 20-fold by exogenous NAD(P)H, and were >85% inhibited by diphenyliodonium (DPI, a flavoenzyme inhibitor). Antagonists of several other potential free radical sources, including xanthine oxidase, nitric oxide synthase, and mitochondrial electron transport, were without effect. Overdistension of intact rabbit iliac arteries in vivo (n = 7) induced 72% fall in intracellular reduced glutathione and 68% increase in oxidized glutathione, so that GSH/GSSG ratio changed from 7.93 +/- 2.14 to 0. 81 +/- 0.16 (p <.005). There was also 28.7% loss of the glutathione pool. Further studies were performed with electron paramagnetic resonance spectroscopy. Rabbit aortas submitted to ex vivo overdistension in the presence of the spin trap DEPMPO (5-diethoxy-phosphoryl-5-methyl-1-pyrroline-N-oxide, 100 mmol/l, n = 5) showed formation of radical adduct spectra, abolished by DPI or superoxide dismutase. Computer simulation indicated a mixture of hydroxyl and carbon-centered radical adducts, likely due to decay of superoxide adduct. Electrical mobility shift assays for NF-kappaB activation were performed in nuclear protein extracts from intact or previously injured rabbit aortas. Balloon injury induced early NF-kappaB activation, which was decreased by DPI. In conclusion, our data show unambiguously that arterial injury induces an immediate profound vascular oxidative stress. Such redox imbalance is likely accounted for by activation of vessel wall NAD(P)H oxidoreductase(s), generating radical species potentially involved in tissue repair. |
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