Monitoring mitochondrial electron fluxes using NAD(P)H-flavoprotein fluorometry reveals complex action of isoflurane on cardiomyocytes.
Monitoring mitochondrial electron fluxes using NAD(P)H-flavoprotein fluorometry reveals complex action of isoflurane on cardiomyocytes.
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DOI：
10.1016/j.bbabio.2010.07.009
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发表时间：
2010-10
影响因子：
4.3
通讯作者：
Bienengraeber, Martin
中科院分区：
文献类型：
--
作者：
Sedlic, Filip;Pravdic, Danijel;Hirata, Naoyuki;Mio, Yasushi;Sepac, Ana;Camara, Amadou K.;Wakatsuki, Tetsuro;Bosnjak, Zeljko J.;Bienengraeber, Martin
关键词：
线粒体生物能研究主要依赖于分离的线粒体，因此排除了对线粒体整体功能重要的其他细胞区室的调节作用。在完整的心肌细胞中，我们通过使用激光扫描共聚焦显微镜同时检测 NAD(H)P 和黄素蛋白 (FP) 荧光强度来跟踪沿电子传递链 (ETC) 特定位点的电子通量动态。该方法用于描述异氟烷（一种挥发性麻醉剂和心脏保护剂）对 ETC 的影响。与充分表征的 ETC 抑制剂和解偶联剂的作用进行比较，揭示了异氟醚的两种不同作用：解偶联诱导的线粒体去极化和在复合物 I 水平上抑制 ETC。相关地，心肌细胞中的耗氧量测量证实了异氟醚的剂量依赖性双重作用，并且在分离的线粒体中，主要在复合物 I 水平上阻碍 ETC。这些作用可能是所报道的轻度刺激线粒体活性氧 (ROS) 产生所需的原因。异氟醚的心脏保护作用。总之，异氟醚对完整心肌细胞中的 ETC 表现出复杂的作用，改变其电子通量，从而增强 ROS 的产生。 NAD(P)H-FP 荧光测定法是探索药物对线粒体的影响并确定其在完整心肌细胞 ETC 内的特定作用位点的有用方法。
Mitochondrial bioenergetic studies mostly rely on isolated mitochondria thus excluding the regulatory role of other cellular compartments important for the overall mitochondrial function. In intact cardiomyocytes, we followed the dynamics of electron fluxes along specific sites of the electron transport chain (ETC) by simultaneous detection of NAD(H)P and flavoprotein (FP) fluorescence intensities using a laser-scanning confocal microscope. This method was used to delineate the effects of isoflurane, a volatile anesthetic and cardioprotective agent, on the ETC. Comparison to the effects of well-characterized ETC inhibitors and uncoupling agent revealed two distinct effects of isoflurane: uncoupling-induced mitochondrial depolarization and inhibition of ETC at the level of complex I. In correlation, oxygen consumption measurements in cardiomyocytes confirmed a dose-dependent, dual effect of isoflurane, and in isolated mitochondria an obstruction of the ETC primarily at the level of complex I. These effects are likely responsible for the reported mild stimulation of mitochondrial reactive oxygen species (ROS) production required for the cardioprotective effects of isoflurane. In conclusion, isoflurane exhibits complex effects on the ETC in intact cardiomyocytes, altering its electron fluxes, and thereby enhancing ROS production. The NAD(P)H-FP fluorometry is a useful method for exploring the effect of drugs on mitochondria and identifying their specific sites of action within the ETC of intact cardiomyocytes.
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影响因子：
2.9
作者：
ESTABROOK, RW
通讯作者：
ESTABROOK, RW
影响因子：
4.8
作者：
Li, NY;Ragheb, K;Robinson, JP
通讯作者：
Robinson, JP
影响因子：
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作者：
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通讯作者：
Bienengraeber, Martin
DOI：
10.1006/bmmb.1993.1051
发表时间：
1993-08-01
期刊：
BIOCHEMICAL MEDICINE AND METABOLIC BIOLOGY
影响因子：
--
作者：
KUNZ, WS;GELLERICH, FN
通讯作者：
GELLERICH, FN
影响因子：
8.8
作者：
Marinovic, J;Bosnjak, ZJ;Stadnicka, A
通讯作者：
Stadnicka, A