Oxygen radical-induced mitochondrial DNA damage and repair in pulmonary vascular endothelial cell phenotypes.
Mitochondrial (mt) DNA is damaged by free radicals. Recent data also show that there are cell type-dependent differences in mtDNA repair capacity. In this study, we explored the effects of xanthine oxidase (XO), which generates superoxide anion directly, and menadione, which enhances superoxide production within mitochondria, on mtDNA in pulmonary arterial (PA), microvascular (MV), and pulmonary venous (PV) endothelial cells (ECs). Both XO and menadione damaged mtDNA in the EC phenotypes, with a rank order of sensitivity of (from most to least) PV > PA > MV for XO and MV = PV > PA for menadione. Dimethylthiourea and deferoxamine blunted menadione- and XO-induced mtDNA damage, thus supporting a role for the iron-catalyzed formation of hydroxyl radical. Damage to the nuclear vascular endothelial growth factor gene was not detected with either XO or menadione. PAECs and MVECs, but not PVECs, repaired XO-induced mtDNA damage quickly. Menadione-induced mtDNA damage was avidly repaired in MVECs and PVECs, whereas repair in PAECs was slower. Analysis of mtDNA lesions at nucleotide resolution showed that damage patterns were similar between EC phenotypes, but there were disparities between XO and menadione in terms of the specific nucleotides damaged. These findings indicate that mtDNA in lung vascular ECs is damaged by XO- and menadione-derived free radicals and suggest that mtDNA damage and repair capacities differ between EC phenotypes.
Grishko
V
V
Department of Pharmacology, College of Medicine, University of South Alabama, Mobile, AL 36688, USA.
Solomon
M
M
Wilson
G L
GL
LeDoux
S P
SP
Gillespie
M N
MN
D000818:Animals; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D004249:DNA Damage / Q000502:physiology*; D004252:DNA Mutational Analysis; D004260:DNA Repair / Q000502:physiology*; D004272:DNA, Mitochondrial / Q000187:drug effects / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D017665:Hydroxyl Radical / Q000378:metabolism; D008833:Microcirculation; D010641:Phenotype; D016133:Polymerase Chain Reaction; D011651:Pulmonary Artery; D011667:Pulmonary Veins; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism; D014812:Vitamin K / Q000378:metabolism / Q000494:pharmacology; D014969:Xanthine Oxidase / Q000378:metabolism / Q000494:pharmacology
D004272:DNA, Mitochondrial; D017382:Reactive Oxygen Species; D013481:Superoxides; D014812:Vitamin K; D017665:Hydroxyl Radical; D014969:Xanthine Oxidase