ENU致大鼠体内Pig-a基因突变的时-效关系和量-效关系研究

doi:10.3969/j.issn.1004-616x.2013.03.008

癌变·畸变·突变

ENU致大鼠体内Pig-a基因突变的时-效关系和量-效关系研究

张铭¹,周长慧¹,王征¹,王庆利²,常艳^{1 ，*}

( 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 )

收稿日期:2012-10-31 修回日期:2012-01-18 出版日期:2013-05-30 发布日期:2013-05-30
通讯作者: 常艳，Tel：021-50800333，E-mail：ychang@ ncdser. com
作者简介:张铭(1986- )，男，山东人，硕士研究生，研究方向：遗传毒理学。E-mail：aiguozhe0301@163.com
基金资助:
国家“十二五”重大专项基金资助项目(2012ZX09505-001-003)

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

ZHANG Ming¹，ZHOU Chang-hui¹，WANG Zheng¹，WANG Qing-li²，CHANG Yan¹，*

(1. China State Institute of Pharmaceutical Industry, National Shanghai Center for New Drug Safety Evaluation & Research, Shanghai 201203; 2. Center for Drug Evaluation, State Food and Drug Administration，Beijing 100038, China)

Received:2012-10-31 Revised:2012-01-18 Online:2013-05-30 Published:2013-05-30
Contact: CHANG Yan，Tel：021-50800333，E-mail：ychang@ ncdser. com

摘要/Abstract

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 glycan class-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 (20 mg/kg)和ENU高剂量组 (40 mg/kg)，灌胃给药，容量均按10 mL/kg，每天1次，连续给药3 d。分别于给药前1 d、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 (red blood cell，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RET Pig-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。

关键词: Pig-a基因, 突变, 时-效关系, 量-效关系

Abstract:

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.

Key words: Pig-a gene, mutation, time-course relationship, dose-response relationship

张铭,周长慧,王征,王庆利,常艳. ENU致大鼠体内Pig-a基因突变的时-效关系和量-效关系研究[J]. 癌变·畸变·突变, doi: 10.3969/j.issn.1004-616x.2013.03.008.

ZHANG Ming，ZHOU Chang-hui，WANG Zheng，WANG Qing-li，CHANG Yan，. Time-course and dose-response relationship study of ENU-induced Pig-a gene mutation in the rat[J]. Carcinogenesis, Teratogenesis & Mutagenesis, doi: 10.3969/j.issn.1004-616x.2013.03.008.

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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

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