The role of singlet oxygen in ischemia-reperfusin injury : the study using laser scanning confocal microscopy and electron paramagnetic resinance

单线态氧在缺血再灌注损伤中的作用：激光扫描共聚焦显微镜和电子顺磁树脂研究

• 批准号: 16390450
• 负责人: ARAI Toshiyuki
• 金额: $ 9.28万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390450/
• 关键词: ischemia-reperfusion injury; singlet oxygen; 6-formylpterin; photodynamic effects; rat hepatocytes; human pancreatic cancer cell line; human leukemia cell line; human dendritic cells; NAC; GSH; ヒスチジン; ヒトT細胞

To elucidate the role of singlet oxygen in ischemia-reperfusion injury, the following studies were performed.1)The generation and quenching of singlet oxygen were chemically examined. When the solution including 6-formylpterin (6FP) was irradiated by fluorescent llight, singlet oxygen was generated, which was quenched by N-acetyl-L-cystein (NAC), glutathine (GSH) and histidine.2)The biological effects of photochemically generated singlet oxygen against were examined using rat hepatocytes. When the hepatocyted were loaded with 6FP and then irradiated by fluorescent llight, the cell death was induced, which was attenuated by the pre-treatment with NAC and GSH, but not with histidine.3)The effects of reactive oxygen species (ROS) generated by 6FP on the drug-induced cell injury were examined. When the rat hepatocytes were loaded with tumor necrosis factor-α and actinomycin D, the apoptotic cell injury was induced, which was attenuated by the pre-treatment with 6FP. The generated ROS by 6FP was thought to suppress the caspase activity and attenuated the apoptotic cell injury.4)The photodynamic effects of a novel pterin derivative were examined using a human pancreatic cancer cell line, Panc-1 cells. When the cells were loaded with the novel pterin and irradiated by ultraviolet (UV-A), the cell death was induced. The singlet oxygen generated during irradiation was thought to cause the cell death.5)The effects of 6FP on the heat-induced apoptosis were examined using a human leukemia cell line, U937 cells. When the cells were loaded with 6FP and exposed to heat shock (44℃ for 20 min), the apoptotic cell death was enhanced. The hydrogen peroxide generated by 6FP was thought to cause the enhancement.6)Lipopolysaccharide (LPS)-induced ROS generation and changes in glutathione level and their relation to the maturation of dendritic cells were examined. It was revealed that the ROS was involved in the cytokine production.

为了阐明单线态氧在缺血再灌注损伤中的作用，进行了以下研究：1)对单线态氧的产生和猝灭进行了化学研究。当含有6-甲酰蝶呤（6FP）的溶液受到荧光光照射时，产生单线态氧，单线态氧被N-乙酰基-L-半胱氨酸（NAC）、谷胱甘肽（GSH）和组氨酸猝灭。2）利用大鼠肝细胞研究光化学产生的单线态氧的生物学效应。当肝细胞负载6FP并用荧光灯照射时，会诱导细胞死亡，NAC和GSH预处理可减轻细胞死亡，但组氨酸则不会减轻这种死亡。3）考察6FP产生的活性氧（ROS）对药物诱导的细胞损伤的影响。当大鼠肝细胞负载肿瘤坏死因子-α和放线菌素D时，会诱导细胞凋亡损伤，6FP预处理可减弱这种损伤。 6FP产生的ROS被认为可以抑制半胱天冬酶活性并减轻细胞凋亡损伤。4)使用人胰腺癌细胞系Panc-1细胞检查了新型蝶呤衍生物的光动力效应。当细胞负载新型蝶呤并受到紫外线（UV-A）照射时，诱导细胞死亡。照射过程中产生的单线态氧被认为是导致细胞死亡的原因。5)使用人白血病细胞系U937细胞检查了6FP对热诱导的细胞凋亡的影响。当细胞负载6FP并热休克（44℃，20分钟）时，细胞凋亡增强。 6FP产生的过氧化氢被认为是导致这种增强的原因。6)研究了脂多糖(LPS)诱导的ROS产生和谷胱甘肽水平的变化及其与树突状细胞成熟的关系。结果表明，ROS 参与了细胞因子的产生。

项目成果

Enhancement of radiation-induced apoptosis by 6-formylpterin

• DOI: 10.1080/1071576042000191754
• 发表时间: 2004-04-01
• 期刊: FREE RADICAL RESEARCH
• 影响因子: 3.3
• 作者: Cui, ZG;Kondo, T;Makino, K
• 通讯作者: Makino, K

Novel 6-formylpterin derivatives: chemical synthesis and O2 to ROS conversion activities.

新型 6-甲酰蝶呤衍生物：化学合成和 O2 到 ROS 的转化活性。

• DOI: 10.1039/b602778d
• 发表时间: 2006
• 期刊: Organic & biomolecular chemistry
• 影响因子: 3.2
• 作者: Mitsuru Nonogawa;T. Arai;N. Endo;S. Pack;T. Kodaki;K. Makino
• 通讯作者: K. Makino

A hydrogen peroxide-generating agent, 6-formylpterin, enhances heat-induced apoptosis

• DOI: 10.1080/02656730400025404
• 发表时间: 2005-05
• 期刊: International Journal of Hyperthermia
• 影响因子: 3.1
• 作者: S. Wada;Zheng-Guo Cui;Takashi Kondo;Qing‐Li Zhao;R. Ogawa;M. Shoji;Toshiyuki Arai;Keisuke Makino;Isao Furuta