Haga Sanae, Remington S James, Morita Naoki, Terui Keita, Ozaki Michitaka
Department of Molecular Surgery, Hokkaido University School of Medicine, Sapporo, Hokkaido, 060-8638 Japan.
Antioxid Redox Signal. 2009 Oct;11(10):2563-72. doi: 10.1089/ars.2009.2681.
Noninvasive evaluation of organ redox states provides invaluable information in many clinical settings. We evaluated a newly developed reduction/oxidation-sensitive green fluorescent protein (roGFP) probe that reports cellular redox potentials and their dynamic changes in live cells. On hypoxia/reoxygenation (H/R) of AML12 liver cells, roGFP indicated mild reduction during hypoxia, but immediate transient oxidation after reoxygenation. The roGFP probe confirmed the antioxidative effects of N-acetylcysteine, catalase, redox factor-1, and Mn-SOD/CuZn-SOD against H/R-induced cellular oxidative stress (OS). In a mouse liver ischemia/reperfusion (I/R) model, roGFP transduced by using an adenoviral vector revealed immediate reduction of the liver under ischemia, and two distinct peaks of OS: (a) early, observed within 60 min after reperfusion, similar to the in vitro study; and (b) later, at 24 h. The early peak levels paralleled the ischemic time up to 75 min and the postischemic liver injury (sGOT/GPT/LDH) in the later phase (6 and 24 h after I/R). The roGFP probe successfully indicated postischemic OS of the liver in living mice, accurately predicting postischemic liver injury. This probe may represent an effective OS marker indicating organ redox states and also predicting the damage/function.
在许多临床环境中，对器官氧化还原状态进行无创评估可提供宝贵信息。我们评估了一种新开发的还原/氧化敏感型绿色荧光蛋白（roGFP）探针，该探针可报告活细胞中的细胞氧化还原电位及其动态变化。在AML12肝细胞进行缺氧/复氧（H/R）处理时，roGFP显示在缺氧期间有轻度还原，但在复氧后立即出现短暂氧化。roGFP探针证实了N-乙酰半胱氨酸、过氧化氢酶、氧化还原因子-1和锰超氧化物歧化酶/铜锌超氧化物歧化酶对H/R诱导的细胞氧化应激（OS）的抗氧化作用。在小鼠肝脏缺血/再灌注（I/R）模型中，通过腺病毒载体转导的roGFP显示缺血时肝脏立即出现还原，以及两个不同的氧化应激峰值：（a）早期，在再灌注后60分钟内观察到，类似于体外研究；（b）后期，在24小时。早期峰值水平与长达75分钟的缺血时间以及后期（I/R后6小时和24小时）的缺血后肝损伤（sGOT/GPT/LDH）平行。roGFP探针成功地显示了活体小鼠肝脏缺血后的氧化应激，准确预测了缺血后肝损伤。该探针可能是一种有效的氧化应激标志物，可指示器官氧化还原状态并预测损伤/功能。
