title: Nitroglycerin induces DNA damage and vascular cell death in the setting of nitrate tolerance.
abstract: Nitroglycerin (GTN) and other organic nitrates are widely used vasodilators. Their side effects are development of nitrate tolerance and endothelial dysfunction. Given the potential of GTN to induce nitro-oxidative stress, we investigated the interaction between nitro-oxidative DNA damage and vascular dysfunction in experimental nitrate tolerance. Cultured endothelial hybridoma cells (EA.hy 926) and Wistar rats were treated with GTN (ex vivo: 10-1000 µM; in vivo: 10, 20 and 50 mg/kg/day for 3 days, s.c.). The level of DNA strand breaks, 8-oxoguanine and O (6)-methylguanine DNA adducts was determined by Comet assay, dot blot and immunohistochemistry. Vascular function was determined by isometric tension recording. DNA adducts and strand breaks were induced by GTN in cells in vitro in a concentration-dependent manner. GTN in vivo administration leads to endothelial dysfunction, nitrate tolerance, aortic and cardiac oxidative stress, formation of DNA adducts, stabilization of p53 and apoptotic death of vascular cells in a dose-dependent fashion. Mice lacking O (6)-methylguanine-DNA methyltransferase displayed more vascular O (6)-methylguanine adducts and oxidative stress under GTN therapy than wild-type mice. Although we were not able to prove a causal role of DNA damage in the etiology of nitrate tolerance, the finding of GTN-induced DNA damage such as the mutagenic and toxic adduct O (6)-methylguanine, and cell death supports the notion that GTN based therapy may provoke adverse side effects, including endothelial function. Further studies are warranted to clarify whether GTN pro-apoptotic effects are related to an impaired recovery of patients upon myocardial infarction.
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Yuliya
authors.initials: Y
authors.lastname: Mikhed
authors.affiliation: Department of Toxicology, University Medical Center of the Johannes Gutenberg University, Obere Zahlbacher Str. 67, 55131, Mainz, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Fahrer
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Oelze
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Swenja
authors.initials: S
authors.lastname: Kröller-Schön
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Steven
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Welschof
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Elena
authors.initials: E
authors.lastname: Zinßius
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Paul
authors.initials: P
authors.lastname: Stamm
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Kashani
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Siyer
authors.initials: S
authors.lastname: Roohani
authors.affiliation: Department of Toxicology, University Medical Center of the Johannes Gutenberg University, Obere Zahlbacher Str. 67, 55131, Mainz, Germany.
authors.forename: Joana Melanie
authors.initials: JM
authors.lastname: Kress
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Ullmann
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Lan P
authors.initials: LP
authors.lastname: Tran
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Eberhard
authors.initials: E
authors.lastname: Schulz
authors.affiliation: Institute of Pharmacy and Biochemistry, Johannes Gutenberg University, Staudinger Weg 5, 55128, Mainz, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Epe
authors.affiliation: Department of Toxicology, University Medical Center of the Johannes Gutenberg University, Obere Zahlbacher Str. 67, 55131, Mainz, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Kaina
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany.|German Center for Cardiovascular Research (DZHK), Partner Site Rhine-Main, Langenbeckstr. 1, 55101, Mainz, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Münzel
authors.affiliation: Center of Cardiology-Cardiology 1, Molecular Cardiology, University Medical Center of the Johannes Gutenberg University, Langenbeckstr. 1, 55101, Mainz, Germany. daiber@uni-mainz.de.|German Center for Cardiovascular Research (DZHK), Partner Site Rhine-Main, Langenbeckstr. 1, 55101, Mainz, Germany. daiber@uni-mainz.de.
authors.forename: Andreas
authors.initials: A
authors.lastname: Daiber
mesh_terms: D000818:Animals; D015153:Blotting, Western; D020552:Comet Assay; D004249:DNA Damage; D004195:Disease Models, Animal; D004361:Drug Tolerance / Q000502:physiology*; D004730:Endothelium, Vascular / Q000187:drug effects*; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D005996:Nitroglycerin / Q000633:toxicity*; D018384:Oxidative Stress; D051381:Rats; D017208:Rats, Wistar; D014665:Vasodilator Agents / Q000633:toxicity*
chemical_list: D014665:Vasodilator Agents; D005996:Nitroglycerin
keywords: DNA damage; Nitrate tolerance; Oxidative stress; Vascular dysfunction