Study on oxidative DNA damage and repair enzyme in acute and chronic renal disease

急慢性肾病中DNA氧化损伤及修复酶的研究

• 批准号: 13671117
• 负责人: HIRAKATA Hideki
• 金额: $ 2.3万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671117/
• 关键词: oxidative DNA damage; 8-oxoguanine; OGG1; acute tubular necrosis; ischemia-reperfusion injury; cisplatin; necrosis; apoptosis; 8-オキソグアニン; 虚血再潜流傷害; necrosis; apoptosis

We examined the contents of 8-oxo-2'-deoxyguanosine (8-oxo-dG), which is a major type of oxidative damage in DNA, in the rat kidney during I/R injury, and also investigated the expression level of the OGG1gene encoding the 8-oxoguanine DNA glycosylase. High-performance liquid chromatography with an MS/MS analysis of the nuclear DNA revealed an immediate accumulation of 8-oxo-dG in the nuclear DNA prepared from the cortex and OM of the kidney 1 h after I/R, and an immunohistochemical analysis demonstrated the immediate accumulation of 8-oxo-dG in the nuclei of renal tubular cells both in the cortex and OM. A delayed increase of cytoplasmic staining with anti-8-oxo-dG was observed only in the cortico-medulla and OM, where the cytoplasmic staining in the proximal tubular cells is higher than in the distal tubular cells. The level of cytoplasmic staining representing 8-oxo-dG in mitochondrial DNA, peaked at 6 h after I/R and preceded the necrosis of proximal tubular cells in the OM, An Rna … More se protection assy showed a high level of OGG1 mRNA in the normal kidney, and the level decreased within 3 h only in the OM, and increased thereafter 1 to 7 days of I/R both in the cortex and OM. In situ hybridization showed higher levels of OGG1 mRNA expression in the renal tubules in the OM than in the cortex of the normal kidney, which decreased rapidly within 3 h of I/R. Thus, the accumulation of 8-oxo-dG in the mitochondrial DNA rather than in nuclear DNA is likely to be involved in the pathogenic responses such as necrosis of renal tubular cells during I/R injury of the kidney, together with an altered level of OGG1 expression.Furthermore, we examined the involvement of 8-oxo-dG in cisplatin-induced renal tubular cell death, in vivo and in vitro. Cisplatin induced accumulation of 8-oxo-dG in renal tubular cells before cell death, and co-administration of DMTU, a scavenger of hydroxyl radicals, inhibited such accumulation and renal tubular damage.In the kidney of 5/6 nephrectomized rats, obvious accumulation of 8-oxo-dG was not observed. Less

我们检测了I/R损伤大鼠肾脏中DNA氧化损伤的主要类型8-氧-2′-脱氧鸟苷（8-氧- dg）的含量，并研究了编码8-氧-鸟嘌呤DNA糖基化酶的ogg1基因的表达水平。核DNA的高效液相色谱和MS/MS分析显示，在I/R后1小时，肾皮质和OM制备的核DNA中立即积累了8-oxo-dG，免疫组织化学分析表明，肾皮质和OM的肾小管细胞细胞核中立即积累了8-oxo-dG。抗8-oxo- dg细胞质染色仅在皮质-髓质和OM中观察到延迟性增加，其中近端小管细胞的细胞质染色高于远端小管细胞。线粒体DNA中代表8-oxo-dG的细胞质染色水平在I/R后6小时达到峰值，在OM近端小管细胞坏死之前，An Rna…More se保护试验显示正常肾脏中OGG1 mRNA水平较高，仅在OM中3小时内水平下降，在I/R后1 ~ 7天皮质和OM中水平均有所上升。原位杂交显示，与正常肾皮质相比，OM肾小管中OGG1 mRNA的表达水平较高，并在I/R后3 h内迅速下降。因此，在肾I/R损伤期间，8-oxo-dG在线粒体DNA中而不是在核DNA中积累可能参与了肾小管细胞坏死等致病反应，并改变了OGG1的表达水平。此外，我们在体内和体外研究了8-oxo-dG在顺铂诱导的肾小管细胞死亡中的作用。顺铂诱导8-氧- dg在细胞死亡前在肾小管细胞中积累，同时给药羟自由基清除剂DMTU可抑制这种积累和肾小管损伤。在5/6肾切除大鼠的肾脏中，未观察到明显的8-oxo-dG积累。少

项目成果

Tsuruya K, et al.: "Accumulation of 8-oxoguanine in the cellular DNA and the alteration of the OGG1 expression during ischemia-reperfusion injury in the rat kidney"DNA Repair. 2. 211-229 (2003)

Tsuruya K 等人：“细胞 DNA 中 8-氧代鸟嘌呤的积累以及大鼠肾脏缺血再灌注损伤期间 OGG1 表达的改变”DNA 修复。