体外培养下氢醌诱导淋巴细胞凋亡及对DNA损伤的影响

目的:研究氢醌对人淋巴细胞的细胞周期阻滞与凋亡、遗传毒性和氧化损伤作用。方法:离体培养淋巴细胞24 h后加S9液,设置氢醌低、中、高浓度(50,150,450μmol/L)组,另设空白对照组,经染毒处理后分别采用MTT比色法检测各组PBL相对存活率,FCM检测细胞周期和凋亡,DCFH-DA法检测ROS含量,FT-IR-ATR检测细胞损伤,SCGE检测DNA链断裂,微核实验检测染色体畸变。结果:氢醌能降低细胞存活度,诱导S+G2/M期周期阻滞,有明显促凋亡作用。细胞内ROS含量剂量依赖性增加,DNA/RNA的光谱区域(1 000～1 490 cm-1)峰型改变明显,高浓度氢醌(450μmol/L)诱导细胞DNA断裂与染色体畸变。结论:氢醌诱导人淋巴细胞结构和功能改变,与DNA氧化应激及DNA-蛋白质损伤有关。

Effects of Hydroquinone on Inducing Apoptosis and DNA Lesion of Lymphocyte in vitro

Objective To investigate the effects of hydroquinone on inducing apoptosis,blocking cell cycles,genotoxicity and oxidative injury of human lymphocyte cells in vitro.Methods Human lymphocyte cells were cultured for 24 hours in vitro and then added S9 solution.The cells were divided into low,middle,high hydroquinone groups with doses of 50,150,450 μmol/L,and control group were set.The relative survival rates of PBL were detected with MTT method,the cell cycle and apoptosis were detected with flow cytometry,the content of ROS were determined with DCFH-DA assay,the cells injury,DNA lesions and chromosome aberration were detected with FT-IR-ATR,SCGE and micronucleus test in all groups after administration,respectively.Results Hydroquinone showed obviously effect on inducing cell apoptosis by decreasing the survival rate and blocking the cell cycle at S and G2/M phase.The contents of ROS was increased in a dose-dependent manner,the variation of DNA/RNA in spectral region from 1 490 to 1 000 cm-1 were apparent,DNA lesions and chromosome aberration were induced with hydroquinone at dose of 450 μmol/L.Conclusion Hydroquinone could induce structure and functional changes in human lymphocyte cells by DNA oxidative stress and lesions.