Mitochondrial dysfunction is an early indicator of doxorubicin-induced apoptosis
Mitochondrial dysfunction is an early indicator of doxorubicin-induced apoptosis
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DOI：
10.1016/s0925-4439(02)00144-8
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发表时间：
2002-10-09
影响因子：
6.2
通讯作者：
Leeuwenburgh, C
中科院分区：
文献类型：
--
作者：
Green, PS;Leeuwenburgh, C
活性氧的产生和线粒体功能障碍与阿霉素诱导的心脏毒性有关。本研究在暴露于阿霉素的H9C2心肌细胞大鼠细胞系和离体大鼠心脏线粒体中检测了促凋亡线粒体死亡信号。使用MTT活力测定法（由功能性线粒体脱氢酶形成的甲醛产物）和钙黄蛋白AM染料（细胞质酯酶裂解后的荧光）评估线粒体和细胞活力。用纳米浓度的阿霉素观察线粒体功能障碍后的细胞死亡。多柔比星诱导的细胞死亡在多柔比星暴露6小时后才出现。多柔比星治疗后，终端（TdT）介导的dutp -生物素缺口末端标记)阳性细胞核的增加证明了细胞凋亡的参与。此外，多柔比星给药分离的线粒体诱导超氧化物产量快速增加，持续至少1小时，随后细胞色素c外排增加。此外，在H9C2心肌细胞系中，caspase-3活性随着阿霉素的使用而增加。阿霉素诱导的细胞凋亡可能需要氧化剂介导的线粒体死亡阈值。(C) 2002 Elsevier Science B.V.版权所有
Generation of reactive oxygen species and mitochondrial dysfunction has been implicated in doxorubicin-induced cardiotoxicity. This study examined pro-apoptotic mitochondrial cell death signals in an H9C2 myocyte rat cell line and in isolated rat heart mitochondria exposed to doxorubicin. Mitochondrial and cellular viability were assessed using an MTT viability assay (formazan product formed by functional mitochondrial dehydrogenases) and calcein AM dye (fluoresces upon cleavage by cytosolic esterases). Mitochondrial dysfunction followed by cell death was observed using nM concentrations of doxorubicin. Significant doxorubicin-induced cell death was not apparent until after 6 h following doxorubicin exposure using the calcein AM assay. The involvement of apoptosis is evidenced by an increase in TUNEL (terminal (TdT)-mediated dUTP-biotin nick end labeling)-positive nuclei following doxorubicin treatment. Furthermore, doxorubicin administered to isolated mitochondria induced a rapid increase in superoxide production, which persisted for at least 1 h and was followed by increased cytochrome c efflux. In addition, caspase-3 activity was increased with doxorubicin administration in the H9C2 myocyte cell line. An oxidant-mediated threshold of mitochondrial death may be required for doxorubicin-induced apoptosis. (C) 2002 Elsevier Science B.V. All rights reserved.