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目的:应用片段化DNA定量分析法和碱性单细胞凝胶电泳法,观察溴化乙锭(EB)对体外培养的小鼠外周血单个核细胞DNA的损伤情况及时间、剂量-效应关系。方法:利用碱性单细胞凝胶电泳法和片段化定量分析法研究在10.0 ng/L EB浓度水平下,培养时间从0～72 h,分析EB对单个核细胞DNA的损伤作用;分别以0、10.0 ng、20.0 ng、50.0 ng/L的EB处理小鼠外周血单个核细胞12 h和24 h,观察剂量-效应关系。结果:10.0 ng/L浓度EB处理的小鼠外周血单个核细胞与对照组在片段化和拖尾率均存在差别(P<0.01);DNA片段化和EB浓度之间存在相关性,24 h时r=0.902(P<0.01);48 h时r=0.819(P<0.05)。结论:溴化乙锭能够明显损伤小鼠外周血单个核细胞DNA,并且呈现明显的时间、剂量效应。
OBJECTIVE: To observe DNA damage and time-dose-response relationship of ethidium bromide (EB) on peripheral blood mononuclear cells cultured in vitro by using fragment DNA quantification and alkaline single cell gel electrophoresis. Methods: The effect of EB on the DNA damage of mononuclear cells was analyzed by alkaline single cell gel electrophoresis and fragmented quantitative analysis at a concentration of 10.0 ng / L EB for 0 ~ 72 h. , 10.0 ng, 20.0 ng and 50.0 ng / L of EB were used to treat peripheral blood mononuclear cells of mice for 12 h and 24 h, and the dose-effect relationship was observed. Results: There was a significant difference in the fragmentation rate and tailing rate between PBMCs treated with 10.0 ng / L EB and control group (P <0.01). The correlation between DNA fragmentation and EB concentration was significant R = 0.902 (P <0.01), and r = 0.819 (P <0.05) at 48 h. Conclusion: Ethidium bromide can obviously damage the DNA of peripheral blood mononuclear cells in mice and show obvious time and dose effects.