Quantitative kinetics of intracellular singlet oxygen generation using a fluorescence probe
Quantitative kinetics of intracellular singlet oxygen generation using a fluorescence probe
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使用荧光探针进行细胞内单线态氧生成的定量动力学
DOI：
10.1038/s41598-020-67155-7
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发表时间：
2020
影响因子：
4.6
通讯作者：
Yoshida Yasukazu
中科院分区：
文献类型：
--
作者：
Murotomi Kazutoshi;Umeno Aya;Sugino Sakiko;Yoshida Yasukazu
单线态氧（1 O2）是一种参与多种生理活动的活性氧。我们以前曾报道过糖尿病前期患者的1 O2特异性氧化产物增加，这表明1 O2的测量可能是生理和病理状况的重要指标。由于1 O2在溶液中的高反应活性和短寿命，在生物活动中1 O2的产生和猝灭的周转非常迅速。然而，1 O2在活细胞中产生的动态变化尚未得到充分的探讨。在这项研究中，我们研究了是否可以使用远红外荧光探针1 O2，Si-DMA，除了1 O2发电机，内过氧化物，哺乳动物细胞1 O2生成的动力学进行定量。用内过氧化物处理哺乳动物活细胞后，Si-DMA荧光强度的动力学呈剂量依赖性增加。另外，处理与1 O2猝灭剂降低荧光强度后，内过氧化物处理。我们的研究结果表明，利用Si-DMA和延时成像技术可以很容易地获得细胞内1 O2的动力学，这为了解哺乳动物细胞中1 O2的产生机制以及1 O2发生剂和猝灭剂的探索提供了新的见解。
Singlet oxygen (1O2) is a type of reactive oxygen species involved in numerous physiological activities. We previously reported that1O2-specific oxidation products are increased in patients with prediabetes, suggesting that measurement of1O2may be an important indicator of physiological and pathological conditions. The turnover in the generation and quenching of1O2is extremely rapid during biological activities owing to it high reactivity and short lifetime in solution. However, the dynamic changes in1O2generation in living cells have not been fully explored. In this study, we investigated whether the kinetics of1O2generation can be quantified using a far-red fluorescent probe for mitochondrial1O2, Si-DMA, following addition of the1O2generator, endoperoxide, to mammalian cells. The kinetics of Si-DMA fluorescence intensity dose-dependently increased following treatment of mammalian living cells with endoperoxide. Alternatively, treatment with1O2quenchers decreased the fluorescence intensities following endoperoxide treatment. Our results indicate that the kinetics of intracellular1O2can be readily obtained using Si-DMA and time-lapse imaging, which provides new insights into the mechanism of1O2generation in mammalian cells and the exploration of1O2generators and quenchers.