On the neurotoxicity mechanism of leukoaminochrome o-semiquinone radical derived from dopamine oxidation:: mitochondria damage, necrosis, and hydroxyl radical formation
On the neurotoxicity mechanism of leukoaminochrome o-semiquinone radical derived from dopamine oxidation:: mitochondria damage, necrosis, and hydroxyl radical formation
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DOI：
10.1016/j.nbd.2004.03.014
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发表时间：
2004-07-01
影响因子：
6.1
通讯作者：
Segura-Aguilar, J
中科院分区：
文献类型：
--
作者：
Arriagada, C;Paris, I;Segura-Aguilar, J
当DT-心肌黄酶被抑制时，多巴胺氧化产物氨基色素单电子还原产生白氨基色素邻半醌自由基。将100 μ M氨基染料与100 μ M双香豆素（DT-心肌黄酶抑制剂）孵育2小时，诱导56%的细胞死亡（P < 0.001）伴随形成（i）细胞内氢过氧化物（与对照相比增加4.2倍; P < 0.001）;（ii）羟基自由基，用ESR和自旋捕获剂检测（与单独的氨基色素相比，当细胞在双香豆素存在下与氨基色素一起孵育时增加2.4倍）;（iii）通过透射电子显微镜测定的细胞内水肿和细胞膜退化;（iv）不存在凋亡，通过使用具有流式细胞术的anexin-V来支持;（v）由荧光染料5，5 ′，6，6 ′-四氯-1，1 ′，3测定的线粒体膜电位的强烈降低，3 ′-四乙基苯并咪唑基碘化乙叉羰（P < 0.01）;（vi）透射电镜观察线粒体内外膜肿胀和破裂。这些结果支持提出的作用leukoaminochrome o-semiquinone自由基作为神经毒素在帕金森病的神经变性和DT-心肌黄酶作为神经保护酶。(C)2004年爱思唯尔公司All rights reserved.
Leukoaminochrome o-semiquinone radical is generated during one-electron reduction of dopamine oxidation product aminochrome when DT-diaphorase is inhibited. Incubation of 100 muM aminochrome with 100 muM dicoumarol, an inhibitor of DT-diaphorase during 2 h, induces 56% cell death (P < 0.001) with concomitant formation of (i) intracellular hydroperoxides (4.2-fold increase compared to control; P < 0.001); (ii) hydroxyl radicals, detected with ESR and spin trapping agents (2.4-fold increase when cells were incubated with aminochrome in the presence of dicoumarol compared to aminochrome alone); (iii) intracellular edema, and cell membrane deterioration determined by transmission electron microscopy; (iv) absence of apoptosis, supported by using anexin-V with flow cytometry; (v) a strong decrease of mitochondrial membrane potential determined by the fluorescent dye 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarboevanineiodide (P < 0.01); (vi) swelling and disruption of outer and inner mitochondrial membranes determined by transmission electron microscopy. These results support the proposed role of leukoaminochrome o-semiquinone radical as neurotoxin in Parkinson's disease neurodegeneration and DT-diaphorase as neuroprotective enzyme. (C) 2004 Elsevier Inc. All rights reserved.