Maruyama W, Kato Y, Yamamoto T, Oh-Hashi K, Hashizume Y, Naoi M
Laboratory of Biochemistry and Metabolism, Department of Basic Gerontology, National Institute for Longevity Sciences, 36-3 Gengo, Morioka-cho, Obu, 474-8522 Aichi.
J Am Aging Assoc. 2001 Jan;24(1):11-8. doi: 10.1007/s11357-001-0002-8.
Peroxynitrite produced from nitric oxide and superoxide has been proposed to cause neuronal dysfunction and cell death in aging and age-related degenerative diseases. 3-Nitrotyosine, an oxidation product of tyrosine by peroxynitrite, was reported to increase in degenerating brains. In this paper, involvement of peroxynitrite in neuronal cell death was studied by analyses of human brains and in vitro experiments on cell death induced by a peroxynitrite-generating agent, SIN-1. 3-Nitrotyrosine-containing proteins were detected in lipofuscin, a typical aging-related pigment in human brains. The cytotoxicity of peroxynitrite was examined in human dopaminergic SH-SY5Y cells by use of SIN-1. SIN-1 induced apoptotic cell death in the cells, and increased the level of 3-nitrotyrosine-containing proteins. The intracellular transduction of death signal was studied in apoptosis induced by peroxynitrite. Apoptosis was induced by sequential death cascade, collapse of mitochondrial membrane potential, activation of caspases and fragmentation of nuclear DNA. In addition, phosphorylation of p38 mitogen activated phosphokinase (MAPK) was found to be associated with apoptosis by SIN-1, as shown by inhibition of apoptotic process by SB202190, a p38 inhibitor. Involvement of peroxynitrite in the cell death is discussed in relation to neuronal degeneration in aging and age-associated diseases.
由一氧化氮和超氧化物产生的过氧亚硝酸盐被认为会在衰老及与年龄相关的退行性疾病中导致神经元功能障碍和细胞死亡。据报道，作为酪氨酸被过氧亚硝酸盐氧化的产物，3-硝基酪氨酸在退化的大脑中含量增加。在本文中，通过对人脑的分析以及对过氧亚硝酸盐生成剂SIN-1诱导的细胞死亡进行体外实验，研究了过氧亚硝酸盐在神经元细胞死亡中的作用。在脂褐素（人脑中一种典型的与衰老相关的色素）中检测到了含3-硝基酪氨酸的蛋白质。利用SIN-1在人多巴胺能SH-SY5Y细胞中检测过氧亚硝酸盐的细胞毒性。SIN-1诱导细胞发生凋亡性细胞死亡，并增加了含3-硝基酪氨酸蛋白质的水平。研究了过氧亚硝酸盐诱导的凋亡中死亡信号的细胞内转导。凋亡是由一系列死亡级联反应、线粒体膜电位崩溃、半胱天冬酶激活和核DNA片段化诱导的。此外，p38丝裂原活化蛋白激酶（MAPK）的磷酸化被发现与SIN-1诱导的凋亡有关，p38抑制剂SB202190对凋亡过程的抑制作用证明了这一点。本文结合衰老和年龄相关疾病中的神经元退化讨论了过氧亚硝酸盐在细胞死亡中的作用。
