Cadenas E, Boveris A, Chance B
Biochem J. 1980 Mar 15;186(3):659-67. doi: 10.1042/bj1860659.
Submitochondrial particles from bovine heart mitochondria showed low-level chemiluminescence when supplemented with organic hydroperoxides. Chemiluminescence seems to measure integratively radical reactions involved in lipid peroxidation and related processes. Maximal light-emission was about 1500 counts/s and was reached 2-10min after addition of hydroperoxides. Ethyl hydroperoxide, cumene hydroperoxide and t-butyl hydroperoxide were effective in that order. Antimycin and rotenone increased chemiluminescence by 50-60%; addition of substrates, NADH and succinate did not produce marked changes in the observed chemiluminescence. Cyanide inhibited chemiluminescence; half-maximal inhibitory effect was obtained with 0.03mm-cyanide and the inhibition was competitive with respect to t-butyl hydroperoxide. Externally added cytochrome c (10-20mum) had a marked stimulatory effect on chemiluminescence, namely a 12-fold increase in light-emission of antimycin-inhibited submitochondrial particles. Stimulation of hydroperoxide-induced chemiluminescence of submitochondrial particles by cytochrome c was matched by a burst of O(2) consumption. O(2) is believed to participate in the chain radical reactions that lead to lipid peroxidation. Superoxide anion seems to be involved in the chemiluminescence reactions as long as light-emission was 50-60% inhibitible by superoxide dismutase. Singlet-oxygen quenchers, e.g. beta-carotene and 1,4-diazabicyclo[2,2,2]-octane, affected light-emission. beta-Carotene was effective either when incorporated into the membranes or added to the cuvette. The present paper suggests that singlet molecular oxygen is mainly responsible for the light-emission in the hydroperoxide-supplemented submitochondrial particles.
牛心线粒体的亚线粒体颗粒在添加有机氢过氧化物时会显示出低水平的化学发光。化学发光似乎综合地测量了脂质过氧化及相关过程中涉及的自由基反应。最大发光量约为1500计数/秒，在添加氢过氧化物后2 - 10分钟达到。乙基氢过氧化物、异丙苯氢过氧化物和叔丁基氢过氧化物按此顺序有效。抗霉素和鱼藤酮使化学发光增加50 - 60%；添加底物、NADH和琥珀酸对观察到的化学发光没有产生明显变化。氰化物抑制化学发光；0.03mmol/L的氰化物可产生半数最大抑制作用，且该抑制作用对叔丁基氢过氧化物具有竞争性。外部添加的细胞色素c（10 - 20μmol）对化学发光有显著的刺激作用，即抗霉素抑制的亚线粒体颗粒的发光增加了12倍。细胞色素c对亚线粒体颗粒氢过氧化物诱导的化学发光的刺激与氧气消耗的爆发相匹配。据信氧气参与导致脂质过氧化的链式自由基反应。只要超氧化物歧化酶可抑制50 - 60%的发光，超氧阴离子似乎就参与化学发光反应。单线态氧猝灭剂，如β - 胡萝卜素和1,4 - 二氮杂双环[2,2,2] - 辛烷，会影响发光。β - 胡萝卜素无论是掺入膜中还是添加到比色皿中都有效。本文表明单线态分子氧主要负责添加氢过氧化物的亚线粒体颗粒中的发光。
