单细胞碱性凝胶电泳用于检测小鼠胸腺细胞DNA链断裂

目的 引进单细胞凝胶电泳(SCGE)实验方法，用以检测小鼠胸腺细胞DNA链断裂。方法 载有经紫外线和H2O2作用过的胸腺细胞和凝胶的玻片首先在pH10的弱碱性裂解液中作用1h，在电泳缓冲液中解旋30min，电泳30min，胸腺细胞核用20mg／L的EB染色5min，用荧光显微镜和显微细胞图像分析系统观察和分析H2O2和紫外线在体外条件下对正常胸腺细胞的DNA损伤作用。结果 紫外线和不同浓度H2O20．001、0．01、0．1、1．0mmoL／L均可导致小鼠胸腺细胞DNA损伤，且随着H2O2浓度的增加DNA损伤逐渐加重。结论 SCGE可用于检测小鼠胸腺细胞DNA链断裂，使用这一实验方法成功地检测出H2O2和紫外线引起的胸腺细胞DNA损伤。

Detection of DNA strand breakage in mice thymus by single-cell gel electropherosis

Objective To establish the single-cell electropherosis (SCGE) protocol for detection of DNA strand breakage in mice thymus. Methods The slides on which thymus cells and agarose were layered were immersed in a cold lysing solution of pH 10 to lyse the thymus cells for 1 h. Then, electrophoresis was performed in electrophoresis running buffer at 30 V and 220 mA for 30 min,the thymus cells nuclei were stained with 20 mg/L of EB for 5 min. The in vitro hydrogen peroxide-induced DNA damage in mice thymus was measured with SCGE established by this experiment. Results The study showed that H 2O 2 and ultraviolet radiation can cause DNA damage. Conclusion SCGE may be used to detect DNA strand breakage in mice thymus. Hydrogen peroxide-induced DNA damage in mice thymus cells is detected successfully using SCGE protocol established by this experiment.