LC-MS/MS分析人支气管上皮细胞与烟曲霉共培养模型中胶霉毒素的含量  

Using LC-MS/MS for determination of Gliotoxin in vitro model of human bronchial epithelial infected by Aspergillus fumigatus

作　　者：沈亮亮[1] 韩禹[1] 郑建锋[1] 赵敬军[1] 

机构地区：[1]同济大学附属同济医院皮肤科,上海200065

出　　处：《中国真菌学杂志》2014年第5期268-271,共4页Chinese Journal of Mycology

摘　　要：目的：采用液相色谱-串联质谱法（LC-MS/MS）分析人支气管上皮细胞与烟曲霉共培养模型中胶霉毒素的含量。方法建立人支气管上皮细胞与烟曲霉共培养模型并于不同时间段检测共培养模型中胶霉毒素的含量。以支气管上皮细胞单独培养为对照组，分别于12h、24h、36h收集对照组、AF293（烟曲霉标准株）共培养组、AFB5233WT（烟曲霉野生株）及AFB5233ΔGlip（烟曲霉胶霉毒素基因敲除株）共培养组细胞培养上清液，采用LC-MS/MS检测胶霉毒素的含量。结果共培养模型中胶霉毒素水平随培养时间增加而逐渐升高（P〈005），胶霉毒素回收率为687%~726%。结论该方法快速、灵敏、结果准确，适用于测定人支气管上皮细胞与烟曲霉共培养模型中胶霉毒素的含量。Objective To establish a LC-MS/MS method for the in vitro determination of Gliotoxin（ GT）in co-culture model of human bronchial epithelial（ HBE）infected by Aspergillus fumigatus（ AF） Methods Developing co-cultured model of HBE infec-ted by AF,the concentration of GT were detected by LC-MS/MS in different periods Results The GT concentration of supernatant from co-culture increased over time（ P 〈0 05）and the recovery of GT was 68 7%-72 6% Conclusion This method is sensi-tive,simple and accurate,and suitable for the determination of GT in the co-culture model.

关 键 词：人支气管上皮细胞 烟曲霉 胶霉毒素 LC-MS/MS 含量测定 

分 类 号：R379.6[医药卫生—病原生物学]