维生素A酸诱导分化的人早幼粒白血病细胞系HL-60的呼吸爆发功能  

Respiratory burst function of the differentiating HL-60 cells induced by retinoic acid

作　　者：任蕴芳[1] 孙存普[2] 丛建波[2] 李秀森[1] 忻文娟[3] 

机构地区：[1]军事医学科学院基础医学研究所 [2]军事医学科学院放射医学研究所,北京100850 [3]中国科学院生物物理研究所

出　　处：《中国药理学与毒理学杂志》1990年第1期49-53,共5页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金

摘　　要：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)。Retinoic acid(RA)induces terminal differentiation of the human promyelocytic leukemia cell line HL-60.The first detectable marker for differentiation is nitroblue terazolium(NBT)reduction that depends upon active oxygen generation in intact cells.In the present paper,TPA-stimulated respiratory burst of HL-60 cells induced by RA or polymorphonuclear(PMN)leukocytes was monitored by using electron spin resonsnce(ESR)method with the spin trap DMPO.The results showed that hydroxyl radicals generated by HL-60 induced by RA and superoxide anions formed by PMN cells were trapped during the respiratory burst course.Furthermore,the experiments with spin probe oxymetry indicated thatthe oxygen in the culture medium was taken up by the cells during the respiratory burst course.Our data suggest that the time course for the respiratory burst of differentiating HL-60 induced by RA was consistent with that for NBT reduction,and the function reached its maximum 3 d after induced.

关 键 词：维生素A酸 诱导分化 HL-60细胞 

分 类 号：R965[医药卫生—药理学]