活性氧在亚砷酸钠诱导Chang肝细胞凋亡中的作用

目的研究在亚砷酸钠（NaAsO2）诱导人Chang肝细胞株的凋亡过程中细胞内活性氧（ROS）和还原型谷胱甘肽（GSH）的作用。方法通过流式细胞术Annexin V/PI双染法检测细胞凋亡,采用2′,7′-二乙酰二氯荧光素（DCFH-DA）检测细胞内ROS水平,应用DTNB法测定细胞内GSH含量。结果与对照组相比,在5～30μmol/LNaAsO2浓度范围内,随着NaAsO2浓度的增高,Chang肝细胞凋亡率、细胞内ROS水平及GSH含量均升高（P〈0.05） 5mmol/L N-乙酰半胱氨酸（NAC）可显著抑制NaAsO2诱导的细胞凋亡的发生（P〈0.05）及细胞内ROS水平的升高（P〈0.05）,但细胞内GSH含量继续升高（P〈0.05）。结论砷诱导的Chang肝细胞的凋亡可能主要取决于细胞内ROS的产生,而不是细胞内GSH含量的下降。

Reactive oxygen species play an important role in NaAsO_2-induced Chang liver cells apoptosis

Objective To study the role of reactive oxygen species（ROS） and reduced glutathione（GSH） in NaAsO2-induced apoptosis in Chang liver cells.Methods The apoptosis rate was assessed by AnnexinV/PI staining.The level of ROS was detected by staining cells with DCFH-DA.The level of intracellular GSH was measured with improved DTNB method.Results After treatment with 5 to 30 μmol/L NaAsO2,compared with control group,a hallmark of apoptosis and accompanied with the enhanced generation of intracellular ROS and GSH were found（P〈0.05）.The addition of N-acetyl-L-cysteine（NAC） effectively rescued the cells from NaAsO2-induced apoptosis and reduced the generation of ROS（P〈0.05）,but elevated the level of GSH（P〈0.05）.Conclusions These data suggest that the arsenic-induced Chang liver cells apoptosis is mostly dependent on generation of ROS rather than GSH depletion.