Enhanced mtDNA repair capacity protects pulmonary artery endothelial cells from oxidant-mediated death

被引:70
作者
Dobson, AW
Grishko, V
LeDoux, SP
Kelley, MR
Wilson, GL
Gillespie, MN [1 ]
机构
[1] Univ S Alabama, Coll Med, Dept Pharmacol, Mobile, AL 36688 USA
[2] Univ S Alabama, Coll Med, Dept Cell Biol & Neurosci, Mobile, AL 36688 USA
[3] Indiana Univ, Sch Med, Wells Ctr Pediat Res, Indianapolis, IN 46202 USA
关键词
mitochondrial deoxyribonucleic acid; xanthine oxidase; Ogg1; cytotoxicity;
D O I
10.1152/ajplung.00443.2001
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
In rat cultured pulmonary arterial (PA), microvascular, and venous endothelial cells (ECs), the rate of mitochondrial (mt) DNA repair is predictive of the severity of xanthine oxidase (XO)-induced mtDNA damage and the sensitivity to XO-mediated cell death. To examine the importance of mtDNA damage and repair more directly, we determined the impact of mitochondrial overexpression of the DNA repair enzyme, Ogg1, on XO-induced mtDNA damage and cell death in PAECs. PAECs were transiently transfected with an Ogg1-mitochondrial targeting sequence construct. Mitochondria-selective overexpression of the transgene product was confirmed microscopically by the observation that immunoreactive Ogg1 colocalized with a mitochondria-specific tracer and, with an oligonucleotide cleavage assay, by a selective enhancement of mitochondrial Ogg1 activity. Overexpression of Ogg1 protected against both XO-induced mtDNA damage, determined by quantitative Southern analysis, and cell death as assessed by trypan blue exclusion and MTS assays. These findings show that mtDNA damage is a direct cause of cell death in XO-treated PAECs.
引用
收藏
页码:L205 / L210
页数:6
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