Mitochondrial DNA damage triggers mitochondrial dysfunction and apoptosis in oxidant-challenged lung endothelial cells.
Oxidant-induced death and dysfunction of pulmonary vascular cells play important roles in the evolution of acute lung injury. In pulmonary artery endothelial cells (PAECs), oxidant-mediated damage to mitochondrial DNA (mtDNA) seems to be critical in initiating cytotoxicity inasmuch as overexpression of the mitochondrially targeted human DNA repair enzyme, human Ogg1 (hOgg1), prevents both mtDNA damage and cell death (Dobson AW, Grishko V, LeDoux SP, Kelley MR, Wilson GL, and Gillespie MN. Am J Physiol Lung Cell Mol Physiol 283: L205-L210, 2002). The mechanism by which mtDNA damage leads to PAEC death is unknown, and the present study tested the specific hypothesis that enhanced mtDNA repair suppresses PAEC mitochondrial dysfunction and apoptosis evoked by xanthine oxidase (XO). PAECs transfected either with an adenoviral vector encoding hOgg1 linked to a mitochondrial targeting sequence or with empty vector were challenged with ascending doses of XO plus hypoxanthine. Quantitative Southern blot analyses revealed that, as expected, hOgg1 overexpression suppressed XO-induced mtDNA damage. Mitochondrial overexpression of hOgg1 also suppressed the XO-mediated loss of mitochondrial membrane potential. Importantly, hOgg1 overexpression attenuated XO-induced apoptosis as detected by suppression of caspase-3 activation, by reduced DNA fragmentation, and by a blunted appearance of condensed, fragmented nuclei. These observations suggest that mtDNA damage serves as a trigger for mitochondrial dysfunction and apoptosis in XO-treated PAECs.
Ruchko
Mykhaylo
M
Department of Pharmacology, University of South Alabama College of Medicine, Mobile, Alabama 36688, USA.
Gorodnya
Olena
O
LeDoux
Susan P
SP
Alexeyev
Mikhail F
MF
Al-Mehdi
Abu-Bakr
AB
Gillespie
Mark N
MN
D000818:Animals; D017209:Apoptosis; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002467:Cell Nucleus / Q000648:ultrastructure; D002478:Cells, Cultured; D004249:DNA Damage; D053938:DNA Fragmentation; D045647:DNA Glycosylases / Q000378:metabolism; D004272:DNA, Mitochondrial; D042783:Endothelial Cells / Q000187:drug effects* / Q000648:ultrastructure; D004789:Enzyme Activation; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology; D011651:Pulmonary Artery / Q000187:drug effects* / Q000503:physiopathology* / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D014969:Xanthine Oxidase / Q000494:pharmacology*
D004272:DNA, Mitochondrial; D016877:Oxidants; D014969:Xanthine Oxidase; D045647:DNA Glycosylases; C483741:OGG1 protein, rat; C505407:CASP3 protein, human; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases