Mitochondrial DNA integrity may be a determinant of endothelial barrier properties in oxidant-challenged rat lungs
Mitochondrial DNA integrity may be a determinant of endothelial barrier properties in oxidant-challenged rat lungs
复制标题
DOI：
10.1152/ajplung.00210.2011
复制
发表时间：
2011-12-01
影响因子：
4.9
通讯作者：
Gillespie, Mark N.
中科院分区：
文献类型：
--
作者：
Chouteau, Joshua M.;Obiako, Boniface;Gillespie, Mark N.
舒托JM，Obiako B，Gorodnya OM，Pastukh VM，Ruchko MV，Wright AJ，Wilson GL，吉莱斯皮MN.线粒体DNA完整性可能是氧化剂激发大鼠肺内皮屏障特性的决定因素。美国生理学杂志肺细胞分子生理学301：L892-L898，2011年。首次发表于2011年9月2日; doi：10.1152/ajplung.00210.2011. -在培养的肺动脉内皮细胞和其他细胞类型中，线粒体靶向DNA修复酶的过表达可保护细胞免受氧化剂诱导的线粒体DNA（mtDNA）损伤和细胞死亡。线粒体DNA的完整性是否支配完整肺循环中内皮细胞的功能特性尚不清楚。因此，本研究使用分离的缓冲液灌注的大鼠肺，以确定是否融合蛋白靶向8-氧代鸟嘌呤DNA糖基化酶1（Ogg1）或核酸内切酶III（Endo III）的线粒体减弱线粒体DNA损伤和血管屏障功能障碍引起的葡萄糖氧化酶（GOX）产生的过氧化氢。我们发现，Endo III和Ogg1融合蛋白在加入灌注介质后30分钟内在肺细胞线粒体中积累。两种结构都防止了GOX诱导的血管滤过系数增加。尽管无法检测到GOX诱导的核DNA损伤，但定量Southern印迹分析显示，GOX诱导的大量氧化性线粒体DNA损伤可以通过两种融合蛋白的预处理来预防。Ogg1结构还逆转了先前存在的GOX诱导的血管屏障功能障碍和氧化mtDNA损伤。总的来说，这些研究结果支持的想法，线粒体DNA是一个哨兵分子管理肺血管屏障反应的氧化应激在完整的肺和线粒体DNA修复途径可能是一个目标的药物干预氧化剂肺损伤。
Chouteau JM, Obiako B, Gorodnya OM, Pastukh VM, Ruchko MV, Wright AJ, Wilson GL, Gillespie MN. Mitochondrial DNA integrity may be a determinant of endothelial barrier properties in oxidant-challenged rat lungs. Am J Physiol Lung Cell Mol Physiol 301: L892-L898, 2011. First published September 2, 2011; doi: 10.1152/ajplung.00210.2011.-In cultured pulmonary artery endothelial cells and other cell types, overexpression of mt-targeted DNA repair enzymes protects against oxidant-induced mitochondrial DNA (mtDNA) damage and cell death. Whether mtDNA integrity governs functional properties of the endothelium in the intact pulmonary circulation is unknown. Accordingly, the present study used isolated, buffer-perfused rat lungs to determine whether fusion proteins targeting 8-oxoguanine DNA glycosylase 1 (Ogg1) or endonuclease III (Endo III) to mitochondria attenuated mtDNA damage and vascular barrier dysfunction evoked by glucose oxidase (GOX)-generated hydrogen peroxide. We found that both Endo III and Ogg1 fusion proteins accumulated in lung cell mitochondria within 30 min of addition to the perfusion medium. Both constructs prevented GOX-induced increases in the vascular filtration coefficient. Although GOX-induced nuclear DNA damage could not be detected, quantitative Southern blot analysis revealed substantial GOX-induced oxidative mtDNA damage that was prevented by pretreatment with both fusion proteins. The Ogg1 construct also reversed preexisting GOX-induced vascular barrier dysfunction and oxidative mtDNA damage. Collectively, these findings support the ideas that mtDNA is a sentinel molecule governing lung vascular barrier responses to oxidant stress in the intact lung and that the mtDNA repair pathway could be a target for pharmacological intervention in oxidant lung injury.