Role of oxidative stress in lysosomal membrane permeabilization and apoptosis induced by gentamicin, an aminoglycoside antibiotic
Role of oxidative stress in lysosomal membrane permeabilization and apoptosis induced by gentamicin, an aminoglycoside antibiotic
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DOI：
10.1016/j.freeradbiomed.2011.07.015
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发表时间：
2011-11-01
影响因子：
7.4
通讯作者：
Mingeot-Leclercq, Marie-Paule
中科院分区：
文献类型：
--
作者：
Denamur, Sophie;Tyteca, Donatienne;Mingeot-Leclercq, Marie-Paule
庆大霉素是一种氨基糖苷类抗生素，用于治疗严重的细菌感染，可能会导致急性肾功能衰竭。在治疗浓度下，庆大霉素聚集在溶酶体中，并诱导肾近端小管细胞凋亡。在庆大霉素处理的肾LLC-PK1细胞中，以前被解释为溶酶体膜通透性的溶酶体释放吖啶橙先于与庆大霉素孵育期间发生的凋亡级联反应。然而，庆大霉素溶酶体蓄积与细胞凋亡之间的联系尚不清楚。我们在这里研究了活性氧(ROS)的产生是否可以解释庆大霉素诱导的丫啶橙释放和细胞凋亡，以及铁在这些事件中的意义。我们发现庆大霉素在吖啶橙释放和细胞凋亡之前诱导ROS的产生，并且在低于需要的药物浓度下诱导ROS的产生。ROS抗氧化剂或清除剂、过氧化氢酶和N-乙酰半胱氨酸在很大程度上阻止了这些事件。生命共聚焦成像显示，庆大霉素诱导的ROS产生发生在溶酶体中。去铁胺是一种铁络合剂，它被内吞并积聚在溶酶体中，在很大程度上阻止了庆大霉素诱导的ROS的产生和细胞凋亡。通过显示荧光黄释放到胞浆中，提供了庆大霉素诱导溶酶体膜通透性的直接证据。荧光黄是一种膜无关的内吞示踪剂，其相对分子质量与庆大霉素相当。总之，我们的数据表明溶酶体铁和早期ROS的产生在庆大霉素诱导的溶酶体膜通透性和细胞凋亡中起着关键作用。(C)2011 Elsevier Inc.保留所有权利。
Gentamicin, an aminoglycoside antibiotic used to treat severe bacterial infections, may cause acute renal failure. At therapeutic concentrations, gentamicin accumulates in lysosomes and induces apoptosis in kidney proximal tubular cells. In gentamicin-treated renal LLC-PK1 cells, acridine orange release from lysosomes, previously interpreted as lysosomal membrane permeabilization, precedes the apoptotic cascade that develops during incubation with gentamicin. However, the link between gentamicin lysosomal accumulation and apoptosis remains unclear. We here examined if reactive oxygen species (ROS) production could account for gentamicin-induced acridine orange release and apoptosis, and the implication of iron in these events. We found that gentamicin induced ROS production prior to, and at lower drug concentrations than required for, acridine orange release and apoptosis. ROS antioxidant or scavenger, catalase, and N-acetylcysteine largely prevented these events. Vital confocal imaging revealed that gentamicin-induced ROS production occurs in lysosomes. Deferoxamine, an iron chelator, which is endocytosed and accumulates in lysosomes, largely prevented gentamicin-induced ROS production as well as apoptosis. Direct evidence for gentamicin-induced permeabilization of lysosomal membrane was provided by showing the release into the cytosol of Lucifer yellow, a membrane-impermeant endocytic tracer with a comparable molecular weight as gentamicin. Altogether, our data demonstrate a key role of lysosomal iron and early ROS production in gentamicin-induced lysosomal membrane permeabilization and apoptosis. (C) 2011 Elsevier Inc. All rights reserved.