Intravital Analysis of Oxidative Stress During Organ Circulatory Failure

器官循环衰竭期间氧化应激的活体分析

• 批准号: 63440033
• 负责人: TSUCHIYA Masaharu
• 金额: $ 9.09万
• 依托单位: Department of Internal Medicine, School of Medicine, Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (A)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63440033/
• 关键词: oxygen radicals; digital imaging; photonic intensifier; microcirculation; chemiluminescence; dichlorofluorescin diacetate; ischemia; oxidative stress; 虚血; 化学発光; dichlorofluorescin; digital image processor; 血小板活性化因子(PAF); 微小循環障害

This project was aimed to establish the methodology to investigate microtopography of oxygen radical formation in microcirculatory units. Recent development of digital imaging photonic microscopy has made it possible to analyze spatial and temporal alterations of oxyradical generation during tissue injury. Luminol-dependent photonic imaging revealed granulocyte-mediated oxidative stress during microvascular damage induced by platelet-activating factor or endotoxin. Our data also demonstrated that the interface between venular indothelium and sticking granulocytes may be the most critical site of oxidant-dependent cellular damage. We have developed another new method to visualize intracellular oxidative stress in the isolated perfused rat liver using digital imaging microfluorography and a hydroperoxide-sensitive fluorescence dye, dichlorofluorescin diacetate. In the experimental model of carbon tetrachloride-induced hepatic injury, the method has revealed that cytochrome P450-dependent oxidant formation takes place predominantly in pericentral regions, preceding cell death at the same area. The current technique will provide a useful method to investigate the microtopograp hy of organ microcirculatory oxidative stress associated with xenobiotic metabolism or circulatory failure.

本研究旨在建立微循环单位氧自由基形成的微地形学研究方法。最近发展的数字成像光子显微镜已经有可能分析组织损伤过程中氧自由基产生的空间和时间变化。鲁米诺依赖性光子成像揭示了血小板活化因子或内毒素诱导的微血管损伤期间粒细胞介导的氧化应激。我们的数据还表明，小静脉吲哚和粘附粒细胞之间的界面可能是氧化剂依赖性细胞损伤的最关键部位。我们已经开发了另一种新的方法来可视化细胞内的氧化应激在离体灌注大鼠肝脏使用数字成像显微荧光和过氧化氢敏感的荧光染料，二氯荧光素二乙酸酯。在四氯化碳诱导的肝损伤的实验模型中，该方法揭示了细胞色素P450依赖性氧化剂的形成主要发生在中央周围区域，在同一区域的细胞死亡之前。该技术为研究外源性代谢或循环衰竭相关的器官微循环氧化应激提供了一种有效的方法。

项目成果

Suematsu, M., Oshio, C., Kurose, I., Miura, S., Asako, H., Sekizuka, E., Houzawa, S,, Nagata, H., Tsuchiya, M: "Profeie analysis of oxyradical diffusion from a single neutrophil using photonic image intensifier microscopy" Cytoprotection and Cytobiology V

Suematsu, M.、Oshio, C.、Kurose, I.、Miura, S.、Asako, H.、Sekizuka, E.、Houzawa, S.、Nagata, H.、Tsuchiya, M：“氧自由基扩散的 Profeie 分析

Suematsu, M., Houzawa, S., Miura. S., Nagata, H., Kitahora, T., Morishita, T., Oshio. C., Tsuchiya, M .: "Effects of serine protease inhibitors on oxyradical burst from phagocytizing neutrophils-Analysis by chemiluminescence counting and its microscopic i

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Asako, H., Suematsu, M., Kato, S., Yanagisawa, T., Ukigai, K., Kurose, I., Suzuki, H., Tsuchiya, M.: "Real-time analysis of oxidative stress in perfused rat hepatic microcirculatory units visualized by digital imaging microscopic fluorography." Microcircu

Asako, H.、Suematsu, M.、Kato, S.、Yanagisawa, T.、Ukigai, K.、Kurose, I.、Suzuki, H.、Tsuchiya, M.：“灌注中氧化应激的实时分析

Suematsu,M.,et.al.: "Intralobular heterogeneity of carbon tetrachlorideーinduced oxidative stress in perfused rat liver visualized by digital imaging fluorescence microscopy." Lab.Invest.64. (1991)

Suematsu,M.,et.al.：“通过数字成像荧光显微镜观察四氯化碳诱导的氧化应激的小叶内异质性”(1991)。

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