三色流式细胞术在小鼠外周血微核检测中的验证  

Validation of the Three-color Flow Cytometry Method for Scoring of Micronucleus in Mouse Peripheral Blood

作　　者：周长慧 王庆利[2] 王征 张铭 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：《中国医药工业杂志》2013年第3期269-273,305,共6页Chinese Journal of Pharmaceuticals

基　　金：十二五重大新药创制滚动平台(2012ZX09302002)

摘　　要：采用不同作用机制的遗传毒性阳性药和阴性药对三色流式细胞术(FCM)检测小鼠外周血微核(MN)进行方法验证,并与常规人工镜检方法对比。每组10只雄性ICR小鼠,分别单次给予溶剂对照(生理盐水或0.5%羧甲纤维素钠)、苯并芘(BaP)、氟尿嘧啶(5-FU)、甲氨蝶呤(MTX)、丝裂霉素C(MMC)、甲磺酸甲酯(MMS)、秋水仙素(Col)、硫酸长春新碱(Vinc)、新化合物A或新化合物B,检测各组小鼠外周血中20 000个网织红细胞(RET)的MN数,同时采用常规显微镜计数各组小鼠骨髓中2 000个嗜多染红细胞(PCE)的MN数。FCM检测的外周血中MN-RET百分率与常规显微镜观察的骨髓中MN-PCE百分率有良好的相关性(r=0.913)。FCM小鼠外周血微核法是一种灵敏、快速、客观的自动化微核检测法,可以作为常规显微镜小鼠骨髓阅片法的替代方法。A study was performed to validate the three-color flow cytometry-based technique （FCM） for enumerating micronucleus （MN） in mouse peripheral blood and to evaluate the degree of correspondence between FCM and conventional microscopy-based method. For these studies, groups of ten mice received a single dose of solvent （normal saline or 0.5 % CMC-Na）, benzo [a]pyrene, fluorouracil, methotrexate, mitomycin C, methyl methanesulfonate, colchicine, vincristine sulfate, new compound A or new compound B. Micronucleated reticulocyte frequencies were measured by FCM E20 000 reticuloeytes （RET） per sample] while micronucleated polychromatic erythrocytes were scored by conventional microscopy-based analysis [2 000 polychromatic erythrocytes （PCE） per sample]. Parallel analysis of micronucleus induction in peripheral blood by FCM and bone marrow using conventional microscopy- based method showed concordant results （r=0.913）. The flow cytometric assessment of MN-RETs in mouse peripheral blood was a sensitive, fast and objective automatic analysis of micronuclei, which could be an acceptable alternative to microscopy-based analysis in mouse bone marrow micronucleus test in vivo.

关 键 词：三色流式细胞术 小鼠外周血 微核 验证 

分 类 号：R99[医药卫生—毒理学]