大鼠体内Pig-a基因突变试验的优化及ENU时-效关系和量-效关系评估

发布时间：2018-09-10 20:06

【摘要】：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。
[Abstract]:Objective to optimize the mutagenesis test of rat Pig-a gene, and to explore the time-effect and dose-effect relationships of the positive mutagens N- ethyl-N-nitrosourea (ENU) in this experiment. Methods Thirty male SD rats were randomly divided into five groups: the solvent control group (n = 10) and the control group (n = 40 mg 路kg ~ (-1) and 80mg/kg (n = 4). The rats were given oral administration of ENU10 mg 路kg ~ (-1) / kg ~ (40 mg / kg) and 80mg/kg for 3 days. The red blood cells were isolated from the tail vein on the 90th day of the experiment. After labeling with Anti-CD59-APC and SYTO 13 nucleic acid dyes, the percentage of CD59 phenotypic mutation to mature erythrocyte (RBCCD59-), CD59 phenotype mutation to reticulocyte (RETCD59-) and the percentage of RET to total erythrocyte were analyzed by flow cytometry. Results the rate of RBCCD59- increased with time and dose reactivity at 1590 days after exposure to ENU. The results showed that the rate of RBCCD59- increased first, then stabilized and fluctuated slightly, but remained at a high level. The peak appeared at 30 days after exposure and decreased at 45 days, and the RETCD59- rate increased with the increase of ENU dose. There was no significant difference in the percentage of RET between the five groups at each time point after 30 days of decrease in the percentage of RET. Conclusion the modified rat Pig-a gene mutation test can be used for rapid detection of Pig-a gene mutation in red blood cells of rats induced by short term (3 d) exposure to Pig-a, which has a time-effect relationship and a dose-effect relationship.
【作者单位】：四川大学华西公共卫生学院四川省食品安全监测与风险评估重点实验室;
【基金】：国家自然科学基金重点项目(No.81030053);国家自然科学基金面上项目(No.81573197)资助
【分类号】：R99

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