MITOCHONDRIAL OXIDATIVE STRESS AFTER CARBON-MONOXIDE HYPOXIA IN THE RAT-BRAIN
MITOCHONDRIAL OXIDATIVE STRESS AFTER CARBON-MONOXIDE HYPOXIA IN THE RAT-BRAIN
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DOI：
10.1172/jci115980
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发表时间：
1992-10-01
影响因子：
15.9
通讯作者：
PIANTADOSI, CA
中科院分区：
文献类型：
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作者：
ZHANG, J;PIANTADOSI, CA
为了更好地理解一氧化碳（CO）缺氧期间和之后的组织损伤机制，我们研究了大鼠脑中部分还原氧（PROS）的产生，该大鼠脑经受1%CO 30 min，然后在空气中再氧合0-180 min。通过测定3-氨基-1，2，4-三唑（ATZ）存在下过氧化氢酶的H2 O2依赖性失活，我们发现复氧后前脑中H2 O2的产生增加。定位的过氧化氢酶的脑microperoxisomes表示的H2 O2生产的细胞内网站，随后的研究中分离的前脑线粒体和CO缺氧后牵连附近的线粒体作为H2 O2的来源。在线粒体中，两个时期的PROS生产的还原型谷胱甘肽/氧化型谷胱甘肽（GSH / GSSG）的比例下降。这些时期的氧化应激发生后立即CO暴露和120分钟后，复氧，GSH/GSSG分别下降50%和43%。谷胱甘肽耗竭数据得到了使用水杨酸探针的羟基自由基生成研究的支持。水杨酸羟基化产物，2，3和2，5-二羟基苯甲酸（DHBA），在线粒体中检测到CO暴露大鼠在GSH/GSSG减少相同的时间间隔内显着增加的金额。DHBA产物在CO暴露后立即增加3.4倍，在120分钟复氧后增加3倍。由于这些氧化应激的迹象并不突出的postmitochondrial分数，我们建议，在大脑中产生的PROS后，CO缺氧主要源于线粒体。这些PROS可能有助于CO介导的神经元损伤严重CO中毒后复氧。
To better understand the mechanisms of tissue injury during and after carbon monoxide (CO) hypoxia, we studied the generation of partially reduced oxygen species (PROS) in the brains of rats subjected to 1% CO for 30 min, and then reoxygenated on air for 0-180 min. By determining H2O2-dependent inactivation of catalase in the presence of 3-amino-1,2,4-triazole (ATZ), we found increased H2O2 production in the forebrain after reoxygenation. The localization of catalase to brain microperoxisomes indicated an intracellular site of H2O2 production; subsequent studies of forebrain mitochondria isolated during and after CO hypoxia implicated nearby mitochondria as the source of H2O2. In the mitochondria, two periods of PROS production were indicated by decreases in the ratio of reduced to oxidized glutathione (GSH / GSSG). These periods of oxidative stress occurred immediately after CO exposure and 120 min after reoxygenation, as indicated by 50 and 43% decreases in GSH/GSSG, respectively. The glutathione depletion data were supported by studies of hydroxyl radical generation using a salicylate probe. The salicylate hydroxylation products, 2,3 and 2,5-dihydroxybenzoic acid (DHBA), were detected in mitochondria from CO exposed rats in significantly increased amounts during the same time intervals as decreases in GSH/GSSG. The DHBA products were increased 3.4-fold immediately after CO exposure, and threefold after 120 min reoxygenation. Because these indications of oxidative stress were not prominent in the postmitochondrial fraction, we propose that PROS generated in the brain after CO hypoxia originate primarily from mitochondria. These PROS may contribute to CO-mediated neuronal damage during reoxygenation after severe CO intoxication.