Yeşilkaya A, Yeğin A, Yücel G, Alicigüzel Y, Aksu T A
Department of Biochemistry, Akdeniz University, School of Medicine, Antalya, Turkey.
Int J Clin Lab Res. 1996;26(1):60-8. doi: 10.1007/BF02644778.
We studied the effect of cumene hydroperoxide, t-butyl hydroperoxide, and hydrogen peroxide on intact healthy human erythrocytes (15 g hemoglobin/dl) using chemiluminescence to monitor peroxidation. We measured the chemiluminescence spectrum, the process of hemolysis, the pH shift, and absorbance spectrum during the incubation with chemicals producing oxidative stress. Maximum chemiluminescence was reached with cumene hydroperoxide at about 50 min, but with t-butyl hydroperoxide at 100 min. The effect of organic hydroperoxide was concentration dependent, whereas the effect of hydrogen peroxide was independent of concentration. Peroxides induced hemolysis after 30 min. The pH shift to alkaline was observed in the first 20-min period. Incubation with organic hydroperoxides induced a decrease in absorption at 580, 545, and 345 nm. Hydrogen peroxide induced a decrease in the same period of time but this returned to the normal range by 120 min. There was no change in absorption at 420 nm with any of the peroxidative agents. Our results suggest that low-level chemiluminescence is a useful model for studying hydroperoxide-induced peroxidation in human erythrocytes.
我们使用化学发光法监测过氧化反应，研究了氢过氧化异丙苯、叔丁基过氧化氢和过氧化氢对完整健康人体红细胞（血红蛋白含量为15 g/dl）的影响。我们测量了与产生氧化应激的化学物质孵育期间的化学发光光谱、溶血过程、pH值变化和吸收光谱。氢过氧化异丙苯在约50分钟时达到最大化学发光，而叔丁基过氧化氢在100分钟时达到最大化学发光。有机过氧化氢的作用呈浓度依赖性，而过氧化氢的作用与浓度无关。过氧化物在30分钟后诱导溶血。在最初的20分钟内观察到pH值向碱性转变。与有机过氧化氢孵育会导致在580、545和345 nm处的吸收降低。过氧化氢在同一时间段内也会导致吸收降低，但在120分钟时恢复到正常范围。使用任何一种过氧化物试剂时，420 nm处的吸收均无变化。我们的结果表明，低水平化学发光是研究过氧化氢诱导人体红细胞过氧化反应的有用模型。
