Reperfusion promotes mitochondrial dysfunction following focal cerebral ischemia in rats.
Reperfusion promotes mitochondrial dysfunction following focal cerebral ischemia in rats.
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DOI：
10.1371/journal.pone.0046498
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Qi S
中科院分区：
文献类型：
--
作者：
Li J;Ma X;Yu W;Lou Z;Mu D;Wang Y;Shen B;Qi S
线粒体功能障碍与脑缺血后观察到的细胞死亡有关，并且已经提出了这种功能障碍的若干机制。短暂性脑缺血后再灌注可引起持续的甚至更严重的脑损伤。许多证据表明，线粒体在缺血性损伤中遭受严重的损伤。本研究旨在观察大鼠局灶性脑缺血再灌注后线粒体功能障碍的特点。雄性Wistar大鼠进行局灶性脑缺血。使用Percoll密度梯度离心分离线粒体。将分离的线粒体固定用于电子显微镜检查;使用分光光度法定量钙诱导的线粒体肿胀。Western blotting检测亲环素D。使用荧光探针选择性染色线粒体，以测量其膜电位，并使用流式细胞术分析来测量活性氧化物质的产生。在暴露于再灌注后的线粒体形态中观察到损伤的迹象。Ca 2+诱导的线粒体肿胀随Ca 2+浓度的增加而逐渐增加，且随再灌注时间的延长而加重。亲环素D蛋白表达在再灌注24小时后达到高峰。线粒体膜电位在再灌注期间显著下降，在再灌注24小时后观察到最大的下降。再灌注2小时后，线粒体活性氧化物质激增，并在再灌注期间维持在较高水平。局灶性脑缺血再灌注引起线粒体形态学损伤和Ca 2+诱导的线粒体肿胀。这种肿胀的机制可能是通过亲环素D蛋白的上调、线粒体膜电位的破坏和过度反应性氧化物质的产生来介导的。
Mitochondrial dysfunction has been implicated in the cell death observed after cerebral ischemia, and several mechanisms for this dysfunction have been proposed. Reperfusion after transient cerebral ischemia may cause continued and even more severe damage to the brain. Many lines of evidence have shown that mitochondria suffer severe damage in response to ischemic injury. The purpose of this study was to observe the features of mitochondrial dysfunction in isolated mitochondria during the reperfusion period following focal cerebral ischemia. Male Wistar rats were subjected to focal cerebral ischemia. Mitochondria were isolated using Percoll density gradient centrifugation. The isolated mitochondria were fixed for electron microscopic examination; calcium-induced mitochondrial swelling was quantified using spectrophotometry. Cyclophilin D was detected by Western blotting. Fluorescent probes were used to selectively stain mitochondria to measure their membrane potential and to measure reactive oxidative species production using flow cytometric analysis. Signs of damage were observed in the mitochondrial morphology after exposure to reperfusion. The mitochondrial swelling induced by Ca2+ increased gradually with the increasing calcium concentration, and this tendency was exacerbated as the reperfusion time was extended. Cyclophilin D protein expression peaked after 24 hours of reperfusion. The mitochondrial membrane potential was decreased significantly during the reperfusion period, with the greatest decrease observed after 24 hours of reperfusion. The surge in mitochondrial reactive oxidative species occurred after 2 hours of reperfusion and was maintained at a high level during the reperfusion period. Reperfusion following focal cerebral ischemia induced significant mitochondrial morphological damage and Ca2+-induced mitochondrial swelling. The mechanism of this swelling may be mediated by the upregulation of the Cyclophilin D protein, the destruction of the mitochondrial membrane potential and the generation of excessive reactive oxidative species.
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影响因子：
2.9
作者：
Hansson, MJ;Persson, T;Elmér, E
通讯作者：
Elmér, E
影响因子：
3.3
作者：
GARCIA, JH
通讯作者：
GARCIA, JH
影响因子：
4.2
作者：
Iijima, T;Mishima, T;Iwao, Y
通讯作者：
Iwao, Y
影响因子：
4.4
作者：
BROWN, AW;BRIERLEY, JB
通讯作者：
BRIERLEY, JB
影响因子：
12.4
作者：
Kowaltowski, A;Vercesi, AE;Fiskum, G
通讯作者：
Fiskum, G