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EFFECT OF INTRACELLULAR REDOX STATUS ON IMMUNOLOGICALLY COMPETENT CELLS : APPLICATION OF THE INDICATOR FOR EVALUATION OF OXIDATIVE STRESS

细胞内氧化还原状态对免疫活性细胞的影响：氧化应激评价指标的应用

基本信息

批准号：
15590118
负责人：
UENO Hitoshi
金额：
$ 1.34万
依托单位：
SETSUNAN 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-15590118/
关键词：
reactive oxidant intermediates redox immune disruption apoptosis ROI carboxy-DCFH-DA hydrogen peroxide flowcytometry

项目摘要

This study was conducted to investigate about relationship between signal transduction in immunologically competent cells and the intracellular redox status that is in a balance on the formation of reactive oxidant intermediates (ROI) and the defense ability against oxidative stress and also to develop an evaluation system for immune disruption by exogenous oxidative stress. When both human cell lines, T-cell leukemia (Jurkat) and promyelocytic leukemia (HL-60) were exposed to hydrogen peroxide (H_2O_2) at the concentration of less than 50 μmol/L that causes mild oxidative stress, ROI was detected by carboxy-DCFH-DA flow cytometry only in the latter cells. This result indicated that there was a difference in redox status between both cell lines even though the same extent of oxidative stress was suffered. In HL-60 cells, viability was decreased by H_2O_2 exposure and this was caused by apoptosis. Production of TNF-α was observed when the cells were exposed to the concentration levels of H_2O_2 that caused apoptosis. Treatment of TNF-α at the detection levels also resulted in the induction of apoptosis. These results suggest that carboxy-DCFH-DA flow cytometry is capable to detect the formation of intracellular ROI which is concomitant with apoptosis induction and TNF-α formation.

本研究旨在探讨免疫活性细胞内信号转导与细胞内氧化还原状态之间的关系，即细胞内氧化还原状态在氧化反应中间体（ROI）的形成和抗氧化应激能力之间的平衡，并建立外源性氧化应激对免疫干扰的评价体系。当人T细胞白血病（Jurkat）和早幼粒细胞白血病（HL-60）细胞暴露于浓度小于50 μmol/L的过氧化氢（H_2O_2）引起轻度氧化应激时，羧基-DCFH-DA流式细胞术仅在后者细胞中检测到ROI。该结果表明，即使遭受相同程度的氧化应激，两种细胞系之间的氧化还原状态也存在差异。H_2O_2暴露可使HL-60细胞活力下降，这是由于细胞凋亡所致。当H_2O_2暴露于引起细胞凋亡的浓度水平时，可观察到TNF-α的产生。检测水平的TNF-α处理也导致细胞凋亡的诱导。这些结果表明，羧基-DCFH-DA流式细胞术能够检测细胞内ROI的形成，其伴随着凋亡诱导和TNF-α的形成。