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Elucidation of internal generation of new nitrogen dioxide-like species and its defense mechanisms.

阐明新的二氧化氮样物质的内部生成及其防御机制。

基本信息

批准号：
11307006
负责人：
OGINO Keiki
金额：
$ 23.8万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

项目摘要

The pathophysiological significance of free-tyrosine or protein-bound tyrosine nitration by the generation of reactive nitrogen species via eosinophil peroxidase was investigated with eosinophils, eosinophil-derived leukemia cells or atopic dermatitis- like-NC/Nga mice. Regarding the immunohistochemical staining method, artifact were occasionally observed in sections containing eosinophils. As a result of this research, an increase in the immunostaining cells for nitrotyrosine in eosinophils of atopic dermatitis-like lesions in NC/Nga mice and the susceptibility of nitrotyrosine staining in eosinophils by reactive nitrogen species derived from adjacent cells were demonstrated. Therefore, it was speculated that eosinophils have a role in the scavenging of reactive nitrogen species. We demonstrated that the expression of myeloperoxidase in human eosinophilic leukemia cell line (Eol-1) occurred when the cells were degenerated by butyric acid and when the ability of nitrotyrosine formation was detected after cytospin preparations of the cells with H_20_2 and NO_2 contrary to the expectation of eosinophil peroxidase expression. Moreover, in further investigations into the localization of peroxidase responsible for the nitration of tyrosine in rat organs, two peroxidases were recognized. One is eosinophil peroxidase found in the gastrointestinal tract and the other is an unknown peroxidase -like that found in the lungs, spleen and heart. In the heart, an unknown peroxidase-like enzymes or the heme protein contributed for the nitration of tyrosine was localized in myocytes.

用嗜酸性粒细胞、嗜酸性粒细胞来源的白血病细胞或异位性皮炎样nc /Nga小鼠研究了游离酪氨酸或蛋白结合酪氨酸硝化通过嗜酸性粒细胞过氧化物酶产生的病理生理意义。免疫组化染色法在含嗜酸性细胞的切片中偶见伪影。本研究结果表明，NC/Nga小鼠特应性皮炎样病变嗜酸性粒细胞中硝基酪氨酸免疫染色细胞增加，邻近细胞衍生的活性氮对嗜酸性粒细胞中硝基酪氨酸染色的敏感性。因此，推测嗜酸性粒细胞在清除活性氮中有一定的作用。结果表明，骨髓过氧化物酶在人嗜酸性白血病细胞系Eol-1中表达的时间与细胞经丁酸变性和H_20_2和NO_2处理后检测到硝基酪氨酸形成能力的时间相反。此外，在对大鼠器官中负责酪氨酸硝化的过氧化物酶定位的进一步研究中，发现了两种过氧化物酶。一种是在胃肠道中发现的嗜酸性过氧化物酶，另一种是一种未知的过氧化物酶，类似于在肺、脾和心脏中发现的过氧化物酶。在心脏中，一种未知的过氧化物酶样酶或血红素蛋白在肌细胞中参与酪氨酸的硝化作用。