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目的研究依他尼酸甲酯(EAME)对人急性髓性白血病HL-60细胞的凋亡诱导作用,并初步探讨EAME诱导HL-60细胞凋亡的机制。方法采用吖啶橙(AO)、溴化乙啶(EB)双染法考察药物的凋亡诱导活性;采用流式细胞术检测活性氧的蓄积和线粒体膜电位的变化;利用荧光标记法检测细胞内还原型谷胱甘肽(GSH)含量的变化。结果EAME浓度在2~10μmol.L-1内对HL-60细胞具有显著的凋亡诱导能力;EAME诱导活性氧蓄积,降低线粒体膜电位和细胞内GSH含量;N-乙酰半胱氨酸(NAC)能够完全逆转EAME对GSH水平的耗竭作用及对HL-60细胞的凋亡诱导作用;丁硫氨酸亚砜胺(BSO)可以协同EAME进一步降低细胞内GSH水平,同时增强凋亡诱导能力。结论EAME可诱导HL-60细胞发生凋亡,活性氧在凋亡诱导过程中起主要作用。细胞内GSH水平与细胞对EAME诱导凋亡的敏感性呈反向相关。
Objective To study the apoptosis-inducing effect of methyl methacrylate (EAME) on human acute myeloid leukemia HL-60 cells and to investigate the mechanism of EAME-induced HL-60 cell apoptosis. Methods Apoptosis-inducing activity of drugs was investigated by acridine orange (AO) and ethidium bromide (EB) staining. The changes of reactive oxygen species accumulation and mitochondrial membrane potential were detected by flow cytometry. Changes in content of reduced glutathione (GSH). Results EAME induced the apoptosis of HL-60 cells in the concentration range of 2 ~ 10μmol.L-1. EAME induced the accumulation of reactive oxygen species, decreased the mitochondrial membrane potential and intracellular GSH content. NAC (NAC ) Completely reversed the effect of EAME on the depletion of GSH and the induction of apoptosis in HL-60 cells. Butylthionine sulfoxide (BSO) synergized with EAME to further reduce intracellular GSH levels and enhance the induction of apoptosis. Conclusion EAME can induce apoptosis in HL-60 cells and reactive oxygen species play a major role in the induction of apoptosis. Intracellular GSH levels were inversely correlated with the sensitivity of cells to EAME-induced apoptosis.