Progression of subcellular changes during chemical hypoxia to cultured rat hepatocytes: A laser scanning confocal microscopic study
Progression of subcellular changes during chemical hypoxia to cultured rat hepatocytes: A laser scanning confocal microscopic study
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DOI：
10.1002/hep.1840210521
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发表时间：
1995-05
期刊：
影响因子：
13.5
通讯作者：
George Zahrebelski;A. Nieminen;K. Al-Ghoul;T. Qian;B. Herman;J. Lemasters
中科院分区：
文献类型：
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作者：
George Zahrebelski;A. Nieminen;K. Al-Ghoul;T. Qian;B. Herman;J. Lemasters
本研究的目的是通过激光扫描共聚焦显微镜评估在氰化物和碘乙酸盐（线粒体呼吸和糖酵解抑制剂）化学缺氧期间培养的肝细胞亚细胞器的变化。使用的参数特异性荧光团是用于细胞形貌和膜通透性的钙黄绿素、用于溶酶体的罗丹明葡聚糖、用于线粒体膜电位 (Δ Ψ) 的罗丹明 123 和四甲基罗丹明甲酯 (TMRM) 以及用于细胞活力损失的碘化丙啶。在培养的肝细胞化学缺氧的前 30 至 40 分钟内，形成大量表面气泡，细胞体积增加，但 Δ Ψ 下降相对较小。随后，线粒体膜的非特异性通透性增加，线粒体去极化。几分钟后，这些事件发生后，单个溶酶体崩解。再过几分钟后，泡破裂、质膜对钙黄绿素的总渗透性和碘化丙啶核标记表明活力丧失。因此，在化学缺氧期间发生了以下一系列细胞内事件：三磷酸腺苷（ATP）耗尽、细胞肿胀形成气泡、线粒体通透性转变开始、溶酶体崩解、气泡破裂导致质膜衰竭和细胞死亡。对缺氧损伤病理生理学的任何解释都必须考虑到这种独特的事件顺序。
The aim of this study was to evaluate changes in the subcellular organelles of cultured hepatocytes by laser scanning confocal microscopy during chemical hypoxia with cyanide and iodoacetate, inhibitors of mitochon‐drial respiration and glycolysis, respectively. Parameter‐specific fluorophores used were calcein for cell topography and membrane permeability, rhodamine‐dextran for lysosomes, rhodamine 123 and tetramethylrhodamine methylester (TMRM) for mitochondrial membrane potential (Δ Ψ) and propidium iodide for loss of cell viability. During the first 30 to 40 minutes of chemical hypoxia to cultured hepatocytes, numerous surface blebs formed and cell volume increased, but Δ Ψ decreased relatively little. Subsequently, the nonspecific permeability of mitochondrial membranes increased, and mitochondria depolarized. These events were followed a few minutes later by disintegration of individual lysosomes. After a few more minutes, viability was lost as indicated by bleb rupture, gross plasma membrane permeability to calcein, and nuclear labeling with propidium iodide. Thus, the following sequence of intracellular events occurred during chemical hypoxia: adenosine triphosphate (ATP) depletion, bleb formation with cellular swelling, onset of a mitochondrial permeability transition, disintegration of lysosomes, plasma membrane failure from bleb rupture, and cell death. Any explanation of the pathophysiology of hypoxic injury must take into account this unique sequence of events.