用吖啶橙-流式细胞仪技术检测间苯二胺、对苯二胺诱导的小鼠骨髓细胞微核率  

Detection of Micronucleus Rate of Mouse Bone Marrow Cells Induced by M-phenylenediamine and P-phenylenediamine with Acriding Organe-Flow Cytometry

作　　者：陈秀娟 江漪 李敏 李培宁 孙侠 陈梓灵 丘智峰 黄宇锋 CHEN Xiu-juan;JIANG Yi;LI-Min;LI-Peining;SUN-Xia;CHEN Zi-ling;Qiu Zhi-feng;HUANG Yu-feng(Guangzhou Quality Supervision and Testing Institute,Guangzhou 511447,China)

机构地区：[1]广州质量监督检测研究院

出　　处：《实验动物与比较医学》2019年第3期231-235,共5页Laboratory Animal and Comparative Medicine

基　　金：广东省质监局科研项目(No.2017CZ04)

摘　　要：目的利用吖啶橙-单激光流式细胞仪(AO-FCM)检测间苯二胺(MPD)、对苯二胺(PPD)的小鼠骨髓细胞微核发生率,建立AO-FCM微核自动化检测方法,探讨FCM在化妆品原料致突变性检测中应用的可行性。方法分别用剂量为25.0 mg/kg、12.5 mg/kg、6.25 mg/kg和3.12 mg/kg的MPD及PPD对小鼠进行处理。采用FCM检测小鼠骨髓中的嗜多染红细胞微核与传统显微镜检测的结果进行比较。结果受试物组小鼠骨髓含微核的嗜多染红细胞数量均有相应的增多,且微核率均随着浓度的增加而增高,呈良好的量效关系。FCM与传统显微镜检测所得的微核率呈良好的相关性(r=0.956,P<0.01)。结论 FCM具有客观性、特异性、高效性,可代替传统显微镜用于检测化妆品原料对小鼠骨髓细胞微核的发生率,评价其致突变性。Objective To detect the micronucleus rate of m-phenylenediamine and p-phenylenediamine in mouse bone marrow cells using the single-laser flow cytometry, and an automated experimental method for rapid detection of micronucleus rate in the laboratory was established to investigate the mutation detection of flow cytometry in cosmetic raw materials. Methods Mice were treated with m-phenylenediamine and p-phenylenediamine at doses of 25.0 mg/kg、12.5 mg/kg、6.25 mg/kg、3.12 mg/kg respectively. Flow cytometry was used to detect the polychromatic erythrocyte micronucleus in mouse bone marrow compared with the results of traditional microscopy. Results The number of Micronucleated polychromatic erythrocytes in the test group was increased correspondingly, and the micronucleus rate was increased with the concentration, showing a good dose-effect relationship. Flow cytometry showed a good correlation with the micronucleus rate obtained by conventional microscopy(r = 0.956, P<0.01). Conclusions Flow cytometric analysis is objectivity, specificity and high efficiency. It can replace the traditional microscope to detect the incidence of cosmetic raw materials on the micronucleus of mouse bone marrow cells and evaluate its mutagenicity.

关 键 词：吖啶橙-流式细胞仪(AO-FCM) 微核试验 骨髓细胞微核率 间苯二胺(MPD) 对苯二胺(PPD) 致突变性 化妆品原料 

分 类 号：Q95-33[生物学—动物学]