tert-Butyl hydroperoxide-induced lipid signaling in hepatocytes:: involvement of glutathione and free radicals
tert-Butyl hydroperoxide-induced lipid signaling in hepatocytes:: involvement of glutathione and free radicals
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DOI：
10.1016/s0006-2952(01)00704-3
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发表时间：
2001-09-15
影响因子：
5.8
通讯作者：
Ruiz-Larrea, MB
中科院分区：
文献类型：
--
作者：
Martín, C;Martínez, R;Ruiz-Larrea, MB
在细胞毒性条件下，叔丁基过氧化氢（TBHP）可从大鼠肝细胞膜磷脂中动员花生四烯酸（AA），从而导致细胞内AA增加，导致细胞死亡。在目前的工作中，参与脂质过氧化，巯基状态，活性氧（ROS）在细胞内AA积累诱导的0.5 MM TBHP在大鼠肝细胞中进行了研究。TBHP处理的细胞保持活力和能量状态在10分钟。然而，TBHP耗尽GSH，以及诱导脂质过氧化和ROS的形成，检测二氯荧光素（DCF）荧光。TBHP还显著增加（32.5%）[C-14]-AA标记的肝细胞内的[C-14]-AA。磷脂酶A（2）（PLA（2））抑制剂米帕林完全抑制[C-14]-AA反应。加入抗氧化剂的细胞悬浮液影响TBHP诱导的脂质反应不同。[C-14]-AA积累与ROS直接相关，与内源GSH负相关。[C-14] -AA与脂质过氧化反应无相关性，异丙嗪可抑制脂质过氧化反应，但不影响[C-14]-AA的增加。我们的结论是，TBHP刺激释放[C-14]-AA从膜磷脂通过PLA（2）介导的机制。内源性GSH和ROS在这种效应中起主要作用，而脂质过氧化相关事件不太可能参与。结果表明，在铁依赖性反应中产生的特定ROS，不同于脂质过氧自由基，参与了PLA（2）的激活，这一过程在TBHP诱导的肝细胞损伤中是重要的。(C)2001 Elsevier Science Inc. All rights reserved.
tert-Butyl hydroperoxide (TBHP) mobilizes arachidonic acid (AA) from membrane phospholipids in rat hepatocytes under cytotoxic conditions, thus leading to an increase in intracellular AA, which precedes cell death. In the present work, the involvement of lipid peroxidation, thiol status, and reactive oxygen species (ROS) in the intracellular AA accumulation induced by 0.5 MM TBHP was studied in rat hepatocytes. Cells treated with TBHP maintained viability and energy status at 10 min. However, TBHP depleted GSH, as well as inducing lipid peroxidation and ROS formation, detected by dichlorofluorescein (DCF) fluorescence. TBHP also significantly increased (32.5%) the intracellular [C-14]-AA from [C-14]-AA-labelled hepatocytes. The phospholipase A(2) (PLA(2)) inhibitor, mepacrine, completely inhibited the [C-14]-AA response. The addition of antioxidants to the cell suspensions affected the TBHP-induced lipid response differently. The [C-14]-AA accumulation correlated directly with ROS and negatively with endogenous GSH. No correlation between [C-14] -AA and lipid peroxidation was found. Promethazine prevented lipid peroxidation and did not affect the [C-14]-AA increase. We conclude that TBHP stimulates the release of [C-14]-AA from membrane phospholipids through a PLA(2)-mediated mechanism. Endogenous GSH and ROS play a major role in this effect, while lipid peroxidation-related events are unlikely to be involved. Results suggest that specific ROS generated in iron-dependent reactions, different from lipid peroxyl radicals, are involved in PLA(2) activation, this process being important in TBHP-induced hepatocyte injury. (C) 2001 Elsevier Science Inc. All rights reserved.