依他尼酸甲酯诱导人急性髓性白血病HL-60细胞凋亡作用和机制的初步探讨  

Apoptotic effect and mechanism of ethacrynic acid methyl ester in human acute myeloid leukemia HL-60 cells

作　　者：王蕊[1] 徐永鹏[1] 赵桂森[2] 赵临襄[3] 景永奎[1] 

机构地区：[1]沈阳药科大学生命科学与生物制药学院,辽宁沈阳110016 [2]山东大学药学院,山东济南250012 [3]沈阳药科大学制药工程学院,辽宁沈阳110016

出　　处：《沈阳药科大学学报》2008年第5期393-398,共6页Journal of Shenyang Pharmaceutical University

基　　金：国家自然科学基金资助项目(30328030,20672069)

摘　　要：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 apoptotic effect and its mechanisms of ethacrynic acid methyl ester （EAME） in HL-60 cells. Methods Apoptotic cells were determined by morphologic observation after AO and EB staining; hydrogen peroxide（H2O2） production and mitochondrial membrane potential（MMP） alteration were determined by flow cytometry after staining with DCFH-DA and Rh123, respectively; intracellular reduced glutathione（GSH） content was determined by fluorometric method after labeling with monochlorobimane. Results EAME significantly induced apoptosis in HL-60 cells at concentrations lower than 10μmol· L^-1. The apoptosis induced by EAME was correlated with H2O2 accumulation and MMP decrease which could be inhibited by catalase. EAME decreased the intracellular GSH levels and NAC blocked EAME-induced the depletion of GSH levels, and apoptosis induction ability. BSO had a synergistic effect with EAME to decrease GSH contents and induce apoptosis in HL-60 cells. Conclusions EAME is a potent apoptosis inducer of HL-60 cells, and EAME-induced apoptosis is mediated through ROS production that can be blocked by increased intracellular GSH levels.

关 键 词：人急性髓性白血病 依他尼酸 依他尼酸甲酯 活性氧 还原型谷胱甘肽 细胞凋亡 

分 类 号：R733[医药卫生—肿瘤] R96[医药卫生—临床医学]