2-甲氧基雌二醇对MUTZ-1细胞内活性氧和线粒体膜电位的影响

Role of reactive oxygen species and mitochondrial membrane potential in 2-methoxyestradiol-induced apoptosis of MUTZ-1 cells

目的探讨在2-甲氧基雌二醇（2-ME）诱导骨髓增生异常综合征（myelodysplastic syndrome，MDS）MUTZ-1细胞凋亡中活性氧（ROS）和线粒体膜电位（△ψm）的作用及其作用机制。方法MUTZ-1细胞与不同浓度2-ME共育，FCM分析荧光探针DCFH-DA标记后细胞内ROS和荧光探针JC-1标记后细胞△ψm的变化。加入抗氧化剂过氧化氢酶（CAT）后，MUTZ-1细胞内相应指标的变化。结果经1-4μmol／L 2-ME作用MUTZ-1细胞12h，与对照组相比，细胞内ROS升高（F=102．56，P〈0.05）和细胞△ψm下调（F=108．02，P〈0．05），且MUTZ-1细胞△ψm下降与细胞内ROS升高成正相关（r=0．981，P=0．019）。CAT能够明显抑制2-ME诱导MUTZ-1细胞内ROS升高（F=68．26，P〈0．01）和细胞△ψm下调（F=55．29，P〈0.01）。结论2-ME诱导MUTZ-1细胞凋亡，可能与2-ME刺激细胞内ROS积聚过多、损伤线粒体结构的完整性以及降低细胞线粒体膜电位有关。

Objective To investigate the changes of reactive oxygen species （ROS） and mitochondrial membrane potential （△ψm） in 2-methoxyestradiol （2-ME）-induced apoptosis of MUTZ-1, a cell line of myelodysplastic syndrome （MDS） , and explore the mechanism. Methods MUTZ-1 cells were incubated with different concentrations （0, 1,2 and 4 ？ tool/L） of 2-ME. The intracellular accumulation of ROS was determined by DCFH-DA staining and the loss of AtOm was detected by flow-cytometry with JC-1 staining, respectively. Results After treatment with 2-ME for 12 hours, an increase of ROS accumulation and loss of AtOm in MUTZ-1 cells were detected in a dose-dependent manner, which were significantly higher than those in control group. There were strong associations between dissipation of △ψm and ROS accumulation at corresponding concentration points （r=0.981 ,P=0.019）. Furthermore, pretreatment with catalase （CAT） significantly rescued the cells from accumulation of ROS（ F = 68.26 ,P 〈 0.01 ） and loss of △ψm（ F = 55. 29 ,P 〈0.01 ）. Conclusion 2-ME-induced apoptosis of MUTZ-1 cells are probably related with intracellular accumulation of ROS and dissipation of △ψm.