Real-time monitoring of reactive oxygen species production during differentiation of human monocytic cell lines (THP-1)
Real-time monitoring of reactive oxygen species production during differentiation of human monocytic cell lines (THP-1)
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DOI：
10.1016/j.aca.2005.06.034
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发表时间：
2005-09-06
影响因子：
6.2
通讯作者：
Yoshimura, T
中科院分区：
文献类型：
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作者：
Kasai, S;Shiku, H;Yoshimura, T
人单核细胞白血病细胞系 THP-1 通过佛波醇肉豆蔻酸酯乙酸酯 (PMA) 处理分化为巨噬细胞。我们报告了分化过程中活性氧（ROS）产生的实时监测。使用电化学和化学发光方法检测了 THP-1 在几个小时内产生的 ROS。在开始用 20 nM-PMA 处理后 1 小时到 10 小时之间，观察到源自 THP-1 的 ROS 的氧化电流增加。与巨噬细胞中观察到的快速氧化反应相比，这种反应异常缓慢。采用化学发光方法进一步研究 20 nM 至 100 KM PMA 处理后的 ROS 产生率。发现用较高浓度的 PMA 处理的细胞的化学发光反应更快。使用化学发光方法监测的氧化反应的时间尺度与使用微电极记录的氧化响应的时间尺度相关。 ROS 产生的缓慢时间尺度反映了 THP-1 向巨噬细胞的分化率。还考察了抑制剂对NADPH氧化酶和NO合酶的影响，发现超氧阴离子是NADPH氧化酶产生的主要自由基。 (c) 2005 Elsevier B.V. 保留所有权利。
Human monocytic leukemia cell line THP-1 differentiates into macrophages by phorbol myristate acetate (PMA) treatment. We report real-time monitoring of reactive oxygen species (ROS) production during the differentiation process. The production of ROS by THP-1 with several hours time scale has been detected using electrochemical and chemiluminescence methods. The increase in oxidation current derived from ROS arising from THP-1 was observed between I and 10 h after the commencement of treatment with 20 nM- PMA. The response was unusually slow compared to the quick oxidative response observed in the macrophages. Chemiluminescence methods were used to further investigate the ROS production rate by carrying out treatment with 20 nM to 100 KM PMA. The chemiluminescence responses were found to be faster for the cells treated with higher concentrations of PMA. The time scale of the oxidative response monitored using the chemiluminescence method correlated with that recorded using a microelectrode. The slow time scale of ROS production reflects the differentiation rate of THP- 1 into macrophages. The effect of inhibitors on NADPH oxidase and NO synthase were also examined to find that the superoxide anion is the main radical produced from NADPH oxidase. (c) 2005 Elsevier B.V. All rights reserved.