ENU致大鼠体内Pig-a基因突变的时-效关系和量-效关系研究  

Time-course and dose-response relationship study of ENU-induced Pig-a gene mutation in the rat

作　　者：张铭 周长慧 王征 王庆利[2] 常艳 

机构地区：[1]中国医药工业研究总院国家上海新药安全评价研究中心,上海201203 [2]国家食品药品监督管理局药品审评中心,北京100038

出　　处：《癌变．畸变．突变》2013年第3期198-200,204,共4页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：国家"十二五"重大专项基金资助项目(2012ZX09505-001-003)

摘　　要：目的：建立大鼠体内Pig-a基因突变试验方法,研究N-乙基-N-亚硝基脲（N-ethyl-N-nitrosourea,ENU）诱导的大鼠体内磷脂酰肌醇聚糖A类（phosphatidylinositol glycanclass-A,Pig-a）基因突变的时-效关系和量-效关系,探索该试验整合到重复剂量毒性试验中的可能性。方法：将15只雄性SD大鼠按照体质量随机分为3组：溶媒对照组（PBS,pH=6.0）、ENU低剂量组（20mg/kg）和ENU高剂量组（40mg/kg）,灌胃给药,容量均按10mL/kg,每天1次,连续给药3d。分别于给药前1d、给药后第15、30和45天颈静脉取血,分离红细胞,经抗体和核酸染料标记后采用流式细胞仪分析网织红细胞（reticulocyte,RET）、总红细胞（redbloodcell,RBC）Pig-a基因突变率和RET百分率。结果：ENU低、高剂量组大鼠在给药后第15、30和45天的RBC和RETPig-a基因突变率平均值与对照组相比均明显升高（P均〈0.01）,高剂量组约为低剂量组的2~3倍。第15~45天,大鼠体内Pig-a基因突变率保持在较高水平,且呈现剂量反应趋势。而在此期间,RET百分率与对照组的比值约为1。结论：本研究成功建立了大鼠体内Pig-a基因突变流式检测方法,提示该试验可整合至重复剂量毒性试验中。OBJECTIVE： To study time-course and dose-response relationship of ENU-induced rat phosphatidylinositol glycan class-A（Pig-a） gene mutation and explore the potential of this assay to be integrated into repeat dose study,and to establish a rat Pig-a gene mutation assay. MEHTODS：Fifteen rats were randomly assigned to be treated with PBS and ENU （20 mg/kg and 40 mg/kg） for 3 consecutive days by oral gavage. Jugular blood samples were collected on days -1 （the day before administration）,15,30 and 45. Erythrocytes were enriched and incubated with Anti-CD59-PE and nucleic acid dye solution,and then analyzed with flow cytometer. RESULTS：On days 15,30 and 45,the Pig-a gene mutation frequencies in RBCs and RETs in low and high dosage groups were elevated significantly compared with that in control group （all P values 0.01）,and the mean frequencies in high dosage group were about 2-3 times of that in low dosage group. The Pig-a gene mutation frequencies were maintained at high levels during days 15-45. CONCLUSION：The in vivo Pig-a gene mutation assay was established successfully in rats. Time-course results suggested that this assay can be integrated into repeated-dose study.

关 键 词：Pig-a基因 突变 时-效关系 量-效关系 

分 类 号：R994.1[医药卫生—毒理学]