Oxidative stress and alterations in actin cytoskeleton trigger glutathione efflux in Saccharomyces cerevisiae
Oxidative stress and alterations in actin cytoskeleton trigger glutathione efflux in Saccharomyces cerevisiae
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DOI：
10.1016/j.bbamcr.2010.07.007
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发表时间：
2010-12-01
影响因子：
5.1
通讯作者：
Vai, Marina
中科院分区：
文献类型：
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作者：
Bradamante, Silvia;Villa, Alessandro;Vai, Marina
谷胱甘肽（GSH）是生命系统中最丰富的抗氧化剂，其显著缺乏在衰老和从神经系统疾病到早期动脉粥样硬化和各种免疫功能受损的疾病的发病机制中起主要作用。在FOTON-M3使命期间，我们进行了空间实验SCORE（酿酒酵母氧化应激反应评价），试图阐明GSH体内平衡。微重力和高氧条件下诱导大量的细胞外释放GSH从S。酿酒酵母细胞（约40%w/dw），改变芽的分布，并激活高渗透压甘油（HOG）和细胞完整性/PKC途径，以及蛋白质羰基化。在模拟微重力条件下进行的一整套实验验证了单次空间飞行实验的结果，结果表明，细胞骨架的改变是所观察到的效应的主要原因。在基础实验中，我们通过用二氢细胞松弛素B（DHC B）（一种有效的肌动蛋白聚合抑制剂）或（R）-（+）-反式-4-（5-氨基-2-甲基-2-苯基）-（5-氨基-2-甲基-2-苯基）-（5-氨基-2-甲基-2-苯基）处理诱导细胞骨架修饰。（1-氨基乙基）-N-（4-吡啶基）环己烷甲酰胺二盐酸盐一水合物（Y-27632），选择性ROCK（Rho相关卷曲螺旋形成蛋白丝氨酸/苏氨酸激酶）抑制剂，证实了肌动蛋白在GSH流出中的作用。我们还发现，GSH的释放可以使用有效的氯离子通道阻断剂5-硝基-2（3-苯基丙基氨基）苯甲酸（NPPB）抑制。(C)2010 Elsevier B. V.保留所有权利。
A marked deficiency in glutathione (GSH), the most abundant antioxidant in living systems, plays a major role in aging and the pathogenesis of diseases ranging from neurological disorders to early atherosclerosis and the impairment of various immunological functions. In an attempt to shed light on GSH homeostasis, we carried out the space experiment SCORE (Saccharomyces cerevisiae oxidative stress response evaluation) during the FOTON-M3 mission. Microgravity and hyperoxic conditions induced an enormous extracellular release of GSH from S. cerevisiae cells (approximate to 40% w/dw), changed the distribution of the buds, and activated the high osmolarity glycerol (HOG) and cell integrity/PKC pathways, as well as protein carbonylation. The results from the single spaceflight experiment were validated by a complete set of experiments under conditions of simulated microgravity and indicate that cytoskeletal alterations are mainly responsible for the observed effects. The results of ground experiments in which we induced cytoskeletal modifications by means of treatment with dihydrocytochalasin B (DHCB), a potent inhibitor of actin polymerisation, or (R)-(+)-trans-4-(1-aminoethyl)-N-(4-pyridyl)cyclohexanecarboxamide dihydrochloride monohydrate (Y-27632), a selective ROCK (Rho-associated coiled-coil forming protein serine/threonine kinase) inhibitor, confirmed the role of actin in GSH efflux. We also found that the GSH release can be inhibited using the potent chloride channel blocker 5-nitro-2(3-phenylpropylamino) benzoic acid (NPPB). (C) 2010 Elsevier B.V. All rights reserved.