Early mitochondrial dysfunction in electron transfer activity and reactive oxygen species generation after cardiac arrest.
Early mitochondrial dysfunction in electron transfer activity and reactive oxygen species generation after cardiac arrest.
复制标题
DOI：
10.1097/ccm.0b013e31818a8a51
复制
发表时间：
2008-11
影响因子：
8.8
通讯作者：
Becker LB
中科院分区：
文献类型：
--
作者：
Han F;Da T;Riobo NA;Becker LB
线粒体生物学似乎是许多进展到死亡的疾病的核心，但在心脏骤停后仍然缺乏特征。缺血时线粒体电子传递功能障碍和活性氧物质（ROS）泄漏可能导致下游事件，包括线粒体蛋白氧化、酪氨酸亚基化、细胞色素c丢失和最终死亡。我们试图更好地定义全动物心脏骤停后这些线粒体功能的早期固定改变。我们使用未经处理的kcl诱导的8分钟心脏骤停，随后复苏和自然循环恢复（ROSC）的小鼠模型，研究四组动物的线粒体功能：(a)心脏骤停8分钟后（CA8）但没有复苏，(b) ROSC后30分钟（R30）， (c) ROSC后60分钟（R60）和(d)假手术。立即收集心脏线粒体，分离并保存在- 80°C，以便稍后使用合适的底物和抑制剂分光光度法测量电子转移活性和ROS泄漏。采用western blot和密度法分析线粒体细胞色素c含量和酪氨酸硝化。在心脏骤停8分钟后，复合体I明显出现ROS渗漏（CA8组，P<0.05），随后复合体I电子传递活性逐渐降低（CA8>R30>R60）。相比之下，复合物II和II - iii活性在评估的时间点表现出更强的抗缺血能力。在酪氨酸硝化过程中观察到~50 kDa和~25 kDa蛋白的早期变化，同时细胞色素c丢失。线粒体功能障碍在缺血和再灌注过程中是一个相对“有序”的过程。随着酪氨酸亚基化和细胞色素c的丢失，线粒体ROS生成和复合体I的电子转移发生变化；这些可能是未来人类治疗的重要新靶点。
Mitochondrial biology appears central to many conditions that progress to death but remains poorly characterized following cardiac arrest. Mitochondrial dysfunction in electron transfer and reactive oxidant species (ROS) leakage during ischemia may lead to downstream events including mitochondrial protein oxidation, tyrosine nitrosylation, cytochrome c loss, and eventual death. We sought to better define early fixed alterations in these mitochondrial functions following whole animal cardiac arrest. We used a murine model of 8 minutes of untreated KCl-induced cardiac arrest followed by resuscitation and return of spontaneous circulation (ROSC) to study mitochondrial functions in four groups of animals: (a) after 8 min cardiac arrest (CA8) but no resuscitation, (b) 30 min post-ROSC (R30), (c) 60 min post-ROSC (R60) and in (d) shams. Heart mitochondria were immediately harvested, isolated and stored at −80°C for later spectrophotometric measurements of electron transfer activities and ROS leakage using appropriate substrates and inhibitors. Mitochondrial cytochrome c content and tyrosine nitration were analyzed by western blot and densitometry. A significant ROS leakage from Complex I was evident after just 8 min of cardiac arrest (CA8 group, P<0.05), which was followed by a progressive reduction in Complex I electron transfer activity (CA8>R30>R60). In contrast, Complex II and II–III activities appeared more resistant to ischemia at the time points evaluated. Early changes in a ~50 kDa and ~25 kDa protein were observed in tyrosine nitration along with a loss of cytochrome c. A relatively “orderly” process of mitochondrial dysfunction progresses during ischemia and reperfusion. Changes in mitochondrial ROS generation and electron transfer from Complex I occur along with tyrosine nitrosylation and loss of cytochrome c; these may represent important new targets for future human therapies.
登录
查看更多内容
DOI：
10.1152/ajpheart.1997.273.3.h1544
发表时间：
1997-09-01
影响因子：
4.8
作者：
Lesnefsky, EJ;Tandler, B;Hoppel, CL
通讯作者：
Hoppel, CL
影响因子：
8.8
作者：
Lavani, Romeen;Chang, Wei-Tien;Vanden Hoek, Terry L.
通讯作者：
Vanden Hoek, Terry L.
影响因子：
4.8
作者：
Poderoso, JJ;Lisdero, C;Boveris, A
通讯作者：
Boveris, A
影响因子：
4.8
作者：
Chen, Q;Vazquez, EJ;Lesnefsky, EJ
通讯作者：
Lesnefsky, EJ
影响因子：
5
作者：
VandenHoek, TL;Li, CQ;Becker, LB
通讯作者：
Becker, LB