DNA damage and repair in cardiovascular disease

心血管疾病中的DNA损伤与修复

• 批准号: 15590749
• 负责人: ISHIDA Mari
• 金额: $ 2.11万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590749/
• 关键词: DNA damage; genomic repair; atherosclerosis

(1)Oxidative stress-induced DNA damage and repair in vascular smooth muscle cells.We found that oxidative stress (H_2O_2) induced the accumulation of thymine glycol and 8-oxo-guanidine in the nucleus, by using immunocytochemical technique. With as little as 30 μM H_2O_2, thymine glycol and 8-oxo-guanidine accumulated in the nucleus 15 minutes after H_2O_2 exposure with a peak at 30 minutes. Upon this oxidative stress-induced DNA damage, DNA repair machinery, such as Rad 51 focus formation and BRCA1 accumulation, was activated. Focus formation of Rad51 did not co-localize with BRCA1 accumulation.(2)Identification of the novel protein(s) that translocate(s) to nucleus by oxidative stress.We found that MAP kinase integrating kinase 1 (Mnk1), which is known to be involed in eukaryotic translation, translocates to nucleus by oxidative stress. Mnk1 is phosphorylated 30 minutes after the exposure to H_2O_2, and co-localized with sc35, showing phosphorylated (activated) Mnk1 is present at the speckles. We are now analyzing the significance of Mnk1 phosphorylation and speckle localization in the nucleus.(3)Polymorphism in genomic repair gene and cardiovascular disease.To examine the association between polymorphism of repair gene and cardiovascular disease, genomic DNA was purified from the blood of patients with or without cardiovascular disease and polymorphism of XRCC3 gene was determined with PCR-RFLP method. About 1000 patients were agreed with the informed consent and DNA analysed. The distribution of the alleles in Japanese seems different from that in Western people. The association between polymorphism of XRCC3 and cardiovascular disease is now being analysed.

(1)氧化应激诱导血管平滑肌细胞DNA损伤和修复。我们利用免疫细胞化学技术发现氧化应激(H_2O_2)诱导胸腺嘧啶乙二醇和8-氧代胍在细胞核内积累。当H_2O_2浓度低至30 μM时，胸腺嘧啶乙二醇和8-氧代胍在H_2O_2暴露后15分钟在细胞核中积累，并在30分钟时达到峰值。在这种氧化应激诱导的 DNA 损伤后，DNA 修复机制（例如 Rad 51 焦点形成和 BRCA1 积累）被激活。 Rad51的焦点形成并不与BRCA1积累共定位。(2)通过氧化应激易位到细胞核的新蛋白质的鉴定。我们发现已知参与真核翻译的MAP激酶整合激酶1(Mnk1)通过氧化应激易位到细胞核。 Mnk1 在暴露于 H_2O_2 后 30 分钟被磷酸化，并与 sc35 共定位，表明斑点处存在磷酸化（激活）的 Mnk1。我们现在正在分析Mnk1磷酸化和细胞核斑点定位的意义。(3)基因组修复基因多态性与心血管疾病。为了研究修复基因多态性与心血管疾病之间的关联，从患有或不患有心血管疾病的患者的血液中纯化基因组DNA，并用XRCC3基因的多态性进行测定。 PCR-RFLP方法。约 1000 名患者签署了知情同意书并进行了 DNA 分析。日本人等位基因的分布似乎与西方人不同。目前正在分析 XRCC3 多态性与心血管疾病之间的关联。

项目成果

Effect of amlodipine and cilazapril treatment on platelet Ca2+ handling in spontaneously hypertensive rats.

• DOI: 10.1291/hypres.26.901
• 发表时间: 2003-11
• 期刊: Hypertension research : official journal of the Japanese Society of Hypertension
• 作者: T. Oshima;N. Ono;R. Ozono;Y. Higashi;M. Ishida;T. Ishida;N. Miho;H. Nakashima;Y. Yano;M. Kambe

Mari Ishida: "Mnk1 is required for angiotensin II-induced protein synthesis in vascular smooth muscle cells"Circulation Research. 93. 1218-1224 (2003)

Mari Ishida：“Mnk1 是血管平滑肌细胞中血管紧张素 II 诱导的蛋白质合成所必需的”循环研究。

Tetsuya Oshima: "Effect of Amlodipine and Cilazapril Treatment on Platelet Ca^<2+> Handling in Spontaneously Hypertensive Rats"Hypertension Research. 26. 901-906 (2003)

Tetsuya Oshima：“氨氯地平和西拉普利治疗对自发性高血压大鼠血小板 Ca^<2> 处理的影响”高血压研究。

Yukiko Nakano: "Matrix Metalloproteinase-9 Contributes to Human Atrial Remodeling During Atrial Fibrillation"Journal of the American College of Cardiology. 43. 818-825 (2004)

Yukiko Nakano：“基质金属蛋白酶-9 有助于房颤期间人类心房重塑”美国心脏病学会杂志。

XRCC3 deficiency results in a defect in recombination and increased endoreduplication in human cells

• DOI: 10.1038/sj.emboj.7600087
• 发表时间: 2004-02
• 期刊: The EMBO Journal
• 作者: Takashi Yoshihara;M. Ishida;A. Kinomura;M. Katsura;Takanori Tsuruga;S. Tashiro;T. Asahara;K. Miyagawa