Mitochondrial ATP production is necessary for activation of the extracellular-signal-regulated kinases during ischaemia/reperfusion in rat myocyte-derived H9c2 cells
Mitochondrial ATP production is necessary for activation of the extracellular-signal-regulated kinases during ischaemia/reperfusion in rat myocyte-derived H9c2 cells
复制标题
DOI：
10.1042/0264-6021:3490119
复制
发表时间：
2000-07-01
影响因子：
4.1
通讯作者：
Guppy, M
中科院分区：
文献类型：
--
作者：
Abas, L;Bogoyevitch, MA;Guppy, M
为了寻找刺激参与激活的丝裂原活化蛋白激酶（MAPKs）在缺血和再灌注期间，我们模拟了一个系统中的事件在体外有利于连续和非侵入性测量的几个主要的扰动发生的时间：O-2张力，线粒体呼吸和能量状态。使用H9 c2细胞（来自大鼠心脏的克隆系），我们发现，激活细胞外信号调节MAPKs（ERK）的复氧被废除，如果线粒体被抑制之前和期间复氧。因此，重新引入O-2 pcv sc不足以激活ERK。通过线粒体呼吸恢复和维持细胞ATP水平是必要的，尽管ATP单独恢复是不够的。ERK激活H2 O2，而不是佛波酯，也敏感的线粒体抑制。因此，复氧和H2 O2介导的氧化应激共享ERK激活的机制，是ATP或ATP依赖性的，这一共同特征表明，复氧反应是由活性氧介导的。ERK活性和ATP水平之间的相关性也被发现在缺氧阶段的缺血，这种影响是不是由于底物限制的激酶。我们的研究结果揭示了细胞代谢在ERK激活中的重要性，并引入ATP作为ERK级联反应机制的新参与者。
To search for the stimuli involved in activating the mitogen-activated protein kinases (MAPKs) during ischaemia and reperfusion, we simulated the event in a system in vitro conducive to continuous and non-invasive measurements of several major perturbations that occur at the time: O-2 tension, mitochondrial respiration and energy status. Using H9c2 cells (a clonal line derived from rat heart), we found that activation of the extracellular signal-regulated MAPKs (ERKs) on reoxygenation was abolished if the mitochondria were inhibited prior to and during reoxygenation. Re-introduction of O-2 pcv sc is therefore not sufficient to activate the ERKs. Recovery and maintenance of cellular ATP levels by mitochondrial respiration is necessary, although ATP recovery alone is not sufficient. ERK activation by H2O2, but not phorbol esters, was also sensitive to mitochondrial inhibition. Thus, reoxygenation and H2O2-mediated oxidative stress share a mechanism of ERK activation that is ATP- or mitochondrion-dependent, and this common feature suggests that the reoxygenation response is mediated by reactive oxygen species. A correlation between ERK activity and ATP levels was also found during the anoxic phase of ischaemia, an effect that was not due to substrate limitation for the kinases. Our results reveal the importance of cellular metabolism in ERK activation, and introduce ATP as a novel participant in the mechanisms underlying the ERK cascade.