Smith J A, Baker M S, Weidemann M J
Division of Biochemistry and Molecular Biology, School of Life Sciences, Faculty of Science, Australian National University, Canberra, ACT.
Biochem Int. 1992 Dec;28(6):1009-20.
Organic hydroperoxides induce oxidative damage to mammalian cells. We describe how luminol-amplified chemiluminescence can be used to monitor free radical generation (following treatment of erythrocytes in vitro with organic hydroperoxides) throughout the entire time-course of oxidative stress. Enrichment of erythrocyte alpha-tocopherol levels increased the induction time by 25% and led peak chemiluminescence fall of 30%. Furthermore, ascorbate loading reduced the signal four-fold during the induction period. The catalytic role of haemoglobin was shown by the abolition of chemiluminescence by azide and a low (but detectable) signal in haemoglobin-depleted erythrocyte ghosts. Luminol-amplified chemiluminescence enables the kinetics of free radical generation to be monitored continuously. Furthermore, it may enable features of the mechanism of interaction between cellular antioxidants and antioxidant enzymes to be elucidated.
有机氢过氧化物会对哺乳动物细胞造成氧化损伤。我们描述了鲁米诺增强化学发光如何用于监测自由基的产生（在体外使用有机氢过氧化物处理红细胞后），贯穿氧化应激的整个时间进程。红细胞α-生育酚水平的升高使诱导时间增加了25%，并导致化学发光峰值下降了30%。此外，在诱导期，抗坏血酸负载使信号降低了四倍。叠氮化物消除化学发光以及血红蛋白耗尽的红细胞空壳中存在低（但可检测到）信号，表明了血红蛋白的催化作用。鲁米诺增强化学发光能够连续监测自由基产生的动力学。此外，它可能有助于阐明细胞抗氧化剂与抗氧化酶之间相互作用机制的特征。
