Giulivi C, Hochstein P, Davies K J
Department of Molecular Pharmacology and Toxicology, University of Southern California, Los Angeles.
Free Radic Biol Med. 1994 Jan;16(1):123-9. doi: 10.1016/0891-5849(94)90249-6.
Red blood cells are frequently employed in studies of oxidative stress. Technical difficulties have previously prevented the measurement of H2O2 production by red blood cells, except during exposure to certain drugs or toxicants. We now show that a combination of glutathione depletion and 3-amino-1,2,4-triazole (aminotriazole) treatment can be used to measure the endogenous generation of H2O2 by red blood cells. In our studies, aminotriazole was used as an H2O2 dependent (irreversible) catalase inhibitor, and catalase inhibition was used as an indirect measure of H2O2 production. Our results indicate that H2O2 is generated at a rate of 1.36 +/- 0.2 microM/h (3.9 +/- 0.6 nmol.h-1.g Hb-1), and that the steady-state red blood cell concentration of H2O2 is approximately 2 x 10(-10) M. Kinetic comparisons of H2O2 production and oxyhemoglobin autooxidation (which generates O2.- that dismutases to H2O2) indicate that the latter is probably the main source of H2O2 in red blood cells.
红细胞常用于氧化应激研究。此前，技术难题阻碍了对红细胞产生过氧化氢（H₂O₂）的测量，只有在接触某些药物或毒物时除外。我们现在表明，谷胱甘肽耗竭与3 - 氨基 - 1,2,4 - 三唑（氨基三唑）处理相结合，可用于测量红细胞内源性产生的H₂O₂。在我们的研究中，氨基三唑用作依赖H₂O₂的（不可逆）过氧化氢酶抑制剂，过氧化氢酶抑制用作H₂O₂产生的间接指标。我们的结果表明，H₂O₂的产生速率为1.36±0.2微摩尔/小时（3.9±0.6纳摩尔·小时⁻¹·克血红蛋白⁻¹），且红细胞中H₂O₂的稳态浓度约为2×10⁻¹⁰摩尔/升。对H₂O₂产生与氧合血红蛋白自动氧化（产生超氧阴离子，超氧阴离子歧化生成H₂O₂）的动力学比较表明，后者可能是红细胞中H₂O₂的主要来源。
