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借助于自旋捕集剂DMPO,用电子自旋共振波谱仪直接定量检测了经维生素A酸(RA)诱导的人早幼粒白血病细胞HL—60受12—O—十四烷酰—佛波醇13—醋酸酯(TPA)刺激引起氧消耗爆发性增加(呼吸爆发)时主要产生的活性氧—羟基自由基(·OH).诱导分化的细胞产生·OH的量随刺激剂TPA在反应体系中量的增加(50～400 ng)而增高,并和RA诱导时间相关,在诱导d3达峰值,和硝基蓝四氮唑(NBT)还原反应及细胞生长曲线的时间过程一致.同时用自旋探针CTPO和TEMPOL及加宽剂Crox现察到RA诱导分化的HL—60细胞呼吸爆发过程中消耗细胞外介质中的氧.同样实验条件下,未经诱导的HL—60细胞不具有呼吸爆发功能,而人多形核白细胞(PMN)呼吸爆发时也消耗细胞外介质中的氧,检测到的活性氧自由基主要为超氧化物阴离子(O_2)。
With the help of the spin-trapping agent DMPO, the electron spin resonance spectroscopy was used to directly detect the concentration of 12-O-tetradecanoyl-phorbol myricetin (HL-60) induced by retinoic acid (RA) in human promyelocytic leukemia cells Alcohol 13-acetate (TPA) stimulates the reactive oxygen species (· OH), which is mainly produced when the oxygen consumption is increased explosively (respiratory burst). The amount of · OH produced by the differentiated cells increases with the stimulus TPA in the reaction system (50-400 ng), and was correlated with the time of RA induction, and reached the peak value at the day of induction of d3, which was consistent with the time course of the reduction reaction of NBT and cell growth curve.At the same time, Spiral probes CTPO and TEMPOL and broadening agent Crox have been shown to elicit depletion of oxygen in the extracellular medium during respiratory burst in RA-induced HL-60 cells.In the same experimental conditions, non-induced HL-60 cells do not have respiration However, PMN also consumes oxygen in the extracellular medium during respiratory burst, and the main active oxygen free radicals detected are superoxide anion (O 2).