Oxidative stress precedes skeletal muscle mitochondrial dysfunction during experimental aortic cross-clamping but is not associated with early lung, heart, brain, liver, or kidney mitochondrial impairment
Oxidative stress precedes skeletal muscle mitochondrial dysfunction during experimental aortic cross-clamping but is not associated with early lung, heart, brain, liver, or kidney mitochondrial impairment
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DOI：
10.1016/j.jvs.2013.07.100
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发表时间：
2014-10-01
影响因子：
4.3
通讯作者：
Geny, Bernard
中科院分区：
文献类型：
--
作者：
Guillot, Max;Charles, Anne-Laure;Geny, Bernard
目的：下肢缺血再灌注会导致骨骼肌线粒体改变、活性氧（ROS）产生和远端器官损伤，这些在很大程度上影响患者的预后。然而，没有再灌注的缺血是否会增加ROS的产生并先于线粒体改变以及线粒体功能障碍是否在远端器官中早期发生尚不清楚。本研究测定了单独缺血后或两期再灌注后的肌肉线粒体功能和 ROS 产生，并研究了下肢缺血再灌注后的心、肺、肝、肾和脑线粒体功能。 方法：将 Wistar 大鼠随机分为四组：假手术组（暴露主动脉但无缺血，n = 9）、I3（仅通过主动脉交叉钳夹 3 小时诱导缺血，n = 9）、 I3R10' 和 I3R2（主动脉阻断，然后再灌注 10 分钟 [n [8] 或 2 小时 [n = 9]）。测量血乳酸、丙氨酸转氨酶、天冬氨酸转氨酶和肌酐。使用克拉克氧电极分析骨骼肌、肺、心脏、脑、肝脏和肾脏中的线粒体呼吸链复合物 I、II、III 和 IV 活性和线粒体耦合（受体控制比）。使用二氢乙锭染色测定肌肉、心脏、肝脏和肾脏中的 ROS 产生。还使用单核细胞-巨噬细胞-2 抗体染色研究了远端器官（心脏、肝脏和肾脏）的炎症。结果：所有组的缺血后乳酸水平均升高。在肌肉中，仅缺血后 ROS 显着增加（+324% +/- 66%；P = .038），再灌注 10 分钟后恢复正常，并在再灌注 2 小时时再次增加（+349.2 +/- 67%；P = .024）。有趣的是，线粒体功能不受单独缺血或随后 10 分钟再灌注的影响，但最大线粒体氧化能力（6.10 +/- 0.51 vs 4.24 +/- 0.36 mu mol/min/g，-30%；P
Objective: Lower limb ischemia-reperfusion results in skeletal muscle mitochondrial alterations, production of reactive oxygen species (ROS), and remote organ impairments that are largely involved in patient prognosis. However, whether ischemia without reperfusion increases ROS production and precedes mitochondrial alteration and whether mitochondrial dysfunction occurs early in remote organs is unknown. This study determined muscle mitochondrial function and ROS production after ischemia alone, or followed by two periods of reperfusion, and investigated heart, lung, liver, kidney, and brain mitochondrial functions after lower limb ischemia-reperfusion.Methods: Wistar rats were randomized into four groups: sham (aortic exposure but no ischemia, n = 9), I3 (ischemia alone induced by aortic cross-clamping for 3 hours, n = 9), I3R10' and I3R2 (aortic cross-clamping, followed by reperfusion for 10 minutes [n [8] or 2 hours [n = 9]). Blood lactate, alanine aminotransferase, aspartate aminotransferase, and creatinine were measured. Mitochondrial respiratory chain complexes I, II, III, and IV activities andmitochondrial coupling (acceptor control ratio) were analyzed using a Clark oxygen electrode in skeletalmuscle, lung, heart, brain, liver, and kidney. ROS production was determined using dihydroethidium staining in muscle, heart, liver, and kidney. Inflammation was also investigated in remote organs (heart, liver, and kidney) using monocyte-macrophage-2 antibody staining.Results: Lactate level increased after ischemia in all groups. In muscle, ROS increased significantly after ischemia alone (+324% +/- 66%; P = .038), normalized after 10 minutes of reperfusion, and increased again at 2 hours of reperfusion (+349.2 +/- 67%; P = .024). Interestingly, mitochondrial function was unaffected by ischemia alone or followed by 10 minutes of reperfusion, but maximal mitochondrial oxidative capacity (6.10 +/- 0.51 vs 4.24 +/- 0.36 mu mol/min/g, -30%; P