论文部分内容阅读
目的对大鼠Pig-a基因突变试验方法进行优化,并初步探讨阳性诱变剂N-乙基-N-亚硝基脲(ENU)在该实验中的时-效关系和量-效关系。方法 30只雄性SD大鼠随机平均分为5组,即溶剂对照组,ENU10mg/kg、20mg/kg、40mg/kg和80mg/kg 4个剂量组,连续3d经口灌胃染毒。分别于实验第0、15、30、45、60、75、90天尾静脉取血,分离红细胞,经Anti-CD59-APC和SYTO 13核酸染料标记后采用流式细胞仪分析CD59表型突变为成熟红细胞(RBCCD59-)率、CD59表型突变为网织红细胞(RETCD59-)率以及RET占总红细胞百分率。结果 ENU各组大鼠在染毒后第15~90天的RBCCD59-率呈现随时间和剂量反应性地增高。RETCD59-率随时间延长,ENU各组表现为先增后基本稳定伴小幅波动趋势,但均保持在较高水平,在染毒后30d出现最高峰,在45d时出现下降;同时,随着ENU剂量的增加,ENU各组RETCD59-率也随之增加。随时间延长,ENU各组大鼠RET百分率有所下降,30d后呈现较为稳定的轻微波动,5组间RET%在各时间点处差异均无统计学意义。结论本研究改良的大鼠Pig-a基因突变试验可进行快速检测,ENU短期(3d)染毒诱导大鼠红细胞Pig-a基因突变具有时-效关系和量-效关系。
OBJECTIVE: To optimize the method for testing the mutation of Pig-a gene in rats and to explore the time-effect and dose-response relationship of the positive mutagen N-ethyl-N-nitrosourea (ENU) in this experiment. Methods Thirty male Sprague-Dawley rats were randomly divided into five groups: solvent control group, ENU 10mg / kg, 20mg / kg, 40mg / kg and 80mg / kg. The blood samples were taken from the tail vein on days 0, 15, 30, 45, 60, 75, and 90, respectively. Erythrocytes were isolated and labeled by Anti-CD59-APC and SYTO 13. The phenotypes of CD59 were analyzed by flow cytometry RBC CD59- rate, RET phenotype of CD59 phenotype, and RET percentage of total erythrocytes. RESULTS: The RBC CD59-rates of rats in ENU groups increased reponsively with time and dose on the 15th to 90th day after exposure. RETCD59-rate over time, ENU showed the first increase after the basic stability with slight fluctuations, but remained at a high level, the peak appeared 30d after exposure, decreased at 45d; the same time, as ENU With the increase of dose, the RETCD59-rate of ENU groups also increased. With the prolongation of time, the percentages of RET in rats in ENU groups decreased slightly and remained stable after 30 days. There was no significant difference in RET% among the five groups at all time points. Conclusion The improved rat Pig-a gene mutation test in this study can be used for rapid detection. There is a time-effect relationship and a dose-response relationship between ENU short-term (3d) exposure induced Pig-a gene mutation in rat erythrocytes.