王蕊,徐永鹏,赵桂森,赵临襄,景永奎
WANG Rui1,XU Yong-peng1,ZHAO Gui-sen2,ZHAO Lin-xiang3,JING Yong-kui1(1.School of Life Science and Biopharmaceutics

• 1:沈阳药科大学生命科学与生物制药学院
• 2:沈阳药科大学生命科学与生物制药学院
• 3:山东大学药学院
• 4:沈阳药科大学制药工程学院

摘要(Abstract)：

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 10 μmol·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(KeyWords)： 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
acute myeloid leukemia;ethacrynic acid;ethacrynic acid methyl ester;reactive oxygen specises;reduced glutathione;apoptosis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(30328030,20672069)

作者(Author): 王蕊,徐永鹏,赵桂森,赵临襄,景永奎
WANG Rui1,XU Yong-peng1,ZHAO Gui-sen2,ZHAO Lin-xiang3,JING Yong-kui1(1.School of Life Science and Biopharmaceutics

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