Analyses of Roles of SOD1 in Protection against Photoaging and Carcinogenesis Using SOD1-deficient Mice

单线态氧对细胞产生劣化作用，并诱导线粒体释放细胞色素c。虽然cyt-c通常诱导凋亡，单线态氧抑制凋亡途径。原因之一是单线态氧诱导cyt-c氧化。单线态氧也使参与细胞凋亡途径的半胱天冬酶和组织蛋白酶失活。尽管DNA和脂质的氧化修饰标记已经建立，但蛋白质的氧化修饰标记还没有合适的。我们检查了四种氨基酸（Trp His、Met和Tyr）的氧化产物，N-甲酰基犬尿氨酸（NFK）是由Trp产生的主要产物。针对NFK的抗体被提出并且被发现与犬尿氨酸（Trp氧化的中间体）以及NFK反应。这是关于抗NFK抗体的首次报道，并且可用于原位检测Trp氧化和犬尿氨酸。我们还建立了一个系统来检测谷胱甘肽化蛋白使用生物素标记的GST作为探针，这使得简单的检测谷胱甘肽化蛋白。我们发现，脾肿大和贫血的SOD 1缺陷小鼠在长期饲养的他们。此外，几乎所有小鼠的面部皮肤都出现了炎症，而抗氧化剂N-乙酰半胱氨酸的使用可以显著延缓炎症的发生。从新生小鼠分离的角质形成细胞的原代培养表明粘附能力下降和培养的血清成分的需要。尽管在常规饲养条件下，SOD 1缺陷小鼠仅报告了次要表型，但长期观察让我们知道，SOD 1缺陷小鼠实际上表现出与老年小鼠相似的症状。

Singlet oxygen exerted deteriorating effects on cells and induced cytochrome c (cyt-c) release from mitochondria. Although cyt-c generally induces apoptosis, singlet oxygen suppressed the apoptotic pathway. One reason is that singlet oxygen induced oxidation of cyt-c. Singlet oxygen also inactivated caspases and cathepsins that are involved in the apoptotic pathway.Although markers for oxidative modification of DNA and lipids are established, there is no proper marker for that of proteins. We examined oxidative products of four amino acids, Trp His, Met, and Tyr, N-formylkynurenine (NFK) was generated from Trp as a major product. Antibody against NFK was raised and found to be reactive to kynurenine, an intermediate of Trp oxidation, as well as NFK. This is the first report on the anti-NFK antibody and could be useful for detection of Trp oxidation and kynurenine in situ. We also established a system to detect glutathionylated proteins using biotinylated GST as a probe, which enabled simple detection of glutathionylated proteins.We found splenomegaly and anemia in SOD1-deficient mice during long-time breeding of them. In addition, almost all mice developed inflammation in facial skin, which is markedly delayed by administration of an antioxidant, N-acetylcysteine. Primary culture of keratinocytes isolated from neonatal mice indicated the decrease in adhering capability and requirement of serum component for culture. Although only minor phenotype has been reported for SOD1-deficient mice under conventional breeding conditions, long-term observation let us know that SOD1-deficient mice actually show symptoms that are similar to aged mice.