A Flow Cytometry-based Assay for Measuring Mitochondrial Membrane Potential in Cardiac Myocytes After Hypoxia/Reoxygenation
A Flow Cytometry-based Assay for Measuring Mitochondrial Membrane Potential in Cardiac Myocytes After Hypoxia/Reoxygenation
复制标题
基于流式细胞术的测定缺氧/复氧后心肌细胞线粒体膜电位的方法
DOI：
10.3791/57725
复制
发表时间：
2018-07-01
影响因子：
1.2
通讯作者：
Gao, Shuo
中科院分区：
文献类型：
--
作者：
Chen, Guihao;Yang, Yuejin;Gao, Shuo
及时有效地对闭塞冠状动脉进行再灌注是减少 ST 段抬高型心肌梗死患者心肌梗死面积的最佳策略。然而，再灌注本身可导致心肌细胞进一步死亡，这种现象称为再灌注损伤。线粒体打开！通透性转换孔（mPTP），随着线粒体的减少！膜电位（MMP），或线粒体！去极化，被普遍认为是再灌注损伤的最后一步，并且与线粒体有关！和心肌细胞死亡。 JC-1 是一种亲脂性阳离子染料，根据 MMP 的值在线粒体中积累。 MMP 越高，JC-1 在线粒体中积累越多。线粒体中 JC-1 含量的增加可以通过荧光发射从绿色（类似于 530 nm）到红色（类似于 590 nm）的变化来反映。因此，红/绿荧光强度比的降低可以表明线粒体的去极化。在这里，我们利用 JC-1 来测量 MMP，即缺氧/复氧后人心肌细胞中 mPTP 的开放，通过流式细胞术检测。
Timely and efficient reperfusion of the occluded coronary artery is the best strategy for decreasing myocardial infarct size in patients with a ST-segment elevated myocardial infarction. However, reperfusion per se can result in further cardiomyocyte death, a phenomenon known as reperfusion injury. The opening of the mitochondria! permeability transition pore (mPTP), with the decrease of the mitochondria! membrane potential (MMP), or mitochondria! depolarization, is universally recognized as the final step of reperfusion injury and is responsible for mitochondria! and cardiomyocyte death. JC-1 is a lipophilic cationic dye that accumulates in mitochondria depending on the value of MMP. The higher the MMP is, the more JC-1 accumulates in the mitochondria. The increasing amounts of JC-1 in mitochondria can be reflected by a fluorescence emission shift from green (similar to 530 nm) to red (similar to 590 nm). Therefore, the reduction of the red/green fluorescence intensity ratio can indicate the depolarization of mitochondria. Here, we take advantage of JC-1 to measure MMP, or the opening of mPTP in human cardiac myocytes after hypoxia/reoxygenation, detected by flow cytometry.