| 基于三色流式细胞术的小鼠外周血微核试验方法的建立 |
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| 引用本文： | 周长慧,王征,王庆利,杨琛懋,常艳. 基于三色流式细胞术的小鼠外周血微核试验方法的建立[J]. 癌变．畸变．突变, 2011, 23(2): 128-133. DOI: 10.3969/j.issn.1004-616x.2011.02.011 |
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| 作者姓名： | 周长慧 王征 王庆利 杨琛懋 常艳 |
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| 作者单位： | 1. 上海医药工业研究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 |
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| 基金项目： | 上海科委专业技术服务能力建设基金资助项目 |
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| 摘 要： | 目的：为克服传统显微镜检微核（micronucleus,MN）方法耗时、费力、主观等缺点,建立基于CD71、CD61荧光抗体和碘化丙啶（cyclophosphamidr,PI）的三色流式细胞术（flow cytometry,FCM）研究小鼠外周血微核的自动化检测方法。方法：经口单次给予小鼠0、20、40和80mg/kg环磷酰胺（CP）,于不同时间点收集外周血;另一批小鼠给予CP40mg/（kg·d）经口灌胃,连续染毒5d,分别于给药前和每次给药后24h收集外周血。采用-80℃甲醇固定血细胞,CD71、CD61荧光抗体、RNA酶和PI孵育血细胞,以疟原虫感染的红细胞作为生物标准物调校FCM后,对CP诱导的小鼠外周血中含MN的CD71＋网织红细胞（reticulocytes,RET）进行检测。结果：单次染毒后48h MN-RET达峰值,且MN-RET呈现明显的剂量依赖性增加（r=0.984 9,P〈0.01）;重复染毒5d,FCM监测的MN-RET染毒2d后达到稳态水平。FCM检测的MN-RET峰值和稳态水平与常规显微镜观察的骨髓微核率有良好的相关性（r=0.9212,P〈0.01）。结论：FCM小鼠外周血微核法稳定可靠,可以作为常规显微镜小鼠骨髓阅片法的替代方法,用于小鼠体内微核试验。
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| 关 键 词： | CD71 流式细胞仪 网织红细胞 微核 环磷酰胺 |
Three-color labeling method for flow cytometric measurement of micronucleus in mouse peripheral blood |
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| ZHOU Chang-hui,WANG Zheng,WANG Qing-li,YANG Chen-mao,CHANG Yan. Three-color labeling method for flow cytometric measurement of micronucleus in mouse peripheral blood[J]. Carcinogenesis,Teratogenesis and Mutagenesis, 2011, 23(2): 128-133. DOI: 10.3969/j.issn.1004-616x.2011.02.011 |
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| Authors: | ZHOU Chang-hui WANG Zheng WANG Qing-li YANG Chen-mao CHANG Yan |
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| Affiliation: | ZHOU Chang-hui~1,WANG Zheng~1,WANG Qing-li~2,YANG Chen-mao~1,CHANG Yan~(1,*) (1.National Shanghai Center for New Drug Safety Evaluation & Research,Shanghai Institute of Pharmaceutical Industry,Shanghai 201203,2.Center for Drug Evaluation,State Food and Drug Administration,Beijing 100038,China) |
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| Abstract: | OBJECTIVE:To overcome time-consuming,laborious,subjective and other shortcomings of the traditional microscopic examination method of micronuclei(MN),we established flow cytometry(FCM) method for automatic analysis of micronuclei in mouse peripheral blood stained with anti-CD71-FITC,anti-CD61-PE and PI. METHOD:Mice received a single oral dose of 0,20,40,80 mg/kg cyclophosphamide and blood was sampled at different times before and after treatment.Additional mice were orally treated with 40 mg/kg cyclophospha... |
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| Keywords: | CD71 |
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