Turrens J F, Giulivi C, Boveris A
J Free Radic Biol Med. 1986;2(2):135-40. doi: 10.1016/s0748-5514(86)80062-9.
The intracellular steady-state concentrations of hydrogen peroxide or superoxide anion were increased by inhibiting either catalase, glutathione peroxidase, or superoxide dismutase activities. Catalase was inhibited with aminotriazole while glutathione peroxidase activity was blocked by eliminating reduced glutathione after addition of either iodoacetamide diethylmaleate or phorone. The concentration of aminotriazole that stimulated chemiluminescence in 50% (60 mM) was very similar to the Ki for catalase activity (70 mM). Cyanide, an inhibitor of both catalase and superoxide dismutase, stimulated chemiluminescence in 50% at a concentration (0.15 mM) which is much closer from the Ki for superoxide dismutase (0.25 mM) than from the Ki for catalase (15 microM). The superoxide dismutase inhibitor diethyldithiocarbamate also increased chemiluminescence six- to ten-fold. Depletion of reduced glutathione stimulated spontaneous chemiluminescence when its concentration decreased below 4.5 mumol X g liver-1. The results shown herein suggest that the changes in the intracellular steady-state concentration occurring after inhibition of any antioxidant enzyme are responsible for the increased spontaneous chemiluminescence. Spontaneous chemiluminescence from intact cells may be used as a noninvasive method for monitoring intracellular free radical metabolism.
通过抑制过氧化氢酶、谷胱甘肽过氧化物酶或超氧化物歧化酶的活性，可提高细胞内过氧化氢或超氧阴离子的稳态浓度。用氨基三唑抑制过氧化氢酶，而在加入碘乙酰胺、马来酸二乙酯或佛尔酮后，通过消除还原型谷胱甘肽来阻断谷胱甘肽过氧化物酶的活性。刺激50%化学发光的氨基三唑浓度（60 mM）与过氧化氢酶活性的Ki值（70 mM）非常相似。氰化物是过氧化氢酶和超氧化物歧化酶的抑制剂，在浓度为0.15 mM时刺激50%的化学发光，该浓度与超氧化物歧化酶的Ki值（0.25 mM）相比，比与过氧化氢酶的Ki值（15 microM）更接近。超氧化物歧化酶抑制剂二乙基二硫代氨基甲酸盐也使化学发光增加了6到10倍。当还原型谷胱甘肽的浓度降至4.5 μmol·g肝脏-1以下时，其耗竭会刺激自发化学发光。本文所示结果表明，抑制任何抗氧化酶后细胞内稳态浓度的变化是自发化学发光增加的原因。完整细胞的自发化学发光可用作监测细胞内自由基代谢的非侵入性方法。
