基于Vero E6细胞的抗SARS药物筛选方法的构建(英文)  

Development of a screening method for potential anti SARS compounds based on Vero-E6 cells

作　　者：王琳[1] 黎荣松[2] 郭海燕[2] 张海清[2] 栗世铀[1] 陈聪[2] 肖培根[3] 谈学海[2] 

机构地区：[1]北京中医药大学中药学院,北京100102 [2]北京华大基因研究中心,北京101300 [3]中国医学科学院中国协和医科大学药用植物研究所,北京100094

出　　处：《中草药》2004年第9期1019-1023,共5页Chinese Traditional and Herbal Drugs

基　　金：国家高技术研究发展计划(863计划)资助项目(2003AA208217)

摘　　要：目的构建基于Vero-E6细胞的抗SARS药物筛选模型。方法运用四唑氮化合物(MTS)方法,对由SARS病毒(BJ-01)引起的对Vero-E6细胞的细胞毒性作用进行检测。首先优化了实验条件(检测了MTS不同孵育时间和不同接种细胞数对测试结果的影响,并绘制了病毒滴度曲线);在检测化合物的抗病毒作用时,首先将细胞接种于96孔板中,加入待测药物和BJ-01病毒,48h后用显微镜观察细胞形态变化,96h后加入MTS和电子耦联剂(PMS)的混合溶液,在490nm检测其吸光度。结果对4000多种化合物的抗SARS病毒作用进行了检测。获得10种可能有抗SARS病毒作用的药物。结论基于Vero-E6细胞的MTS检测方法提供了一种快速、安全和方便的抗SARS药物筛选方法。Object To develop a Vero E6 cell line based assay for screening potential anti SARS compounds. Methods Conditions of the Vero E6 cell line based MTS assay were optimized. Virus titer curve had been generated. To screen anti SARS virus compounds, Vero E6 cells were plated in 96 well plates, test compounds were added and immediately followed with BJ 01 SARS virus. Cell morphology was observed by microscope after 48 h of incubation. MTS and PMS mixture was added in 96 h and absorbance values were measured at 490 nm. Results More than 4 000 compounds had been screened and ten potential anti SARS compounds had been found. Conclusion The Vero E6 cell based MTS assay offers a rapid, safe, and convenient method for screening potential anti SARS compounds.

关 键 词：重急性呼吸综合征 VERO-E6细胞 四唑氮化合物 

分 类 号：R965.1[医药卫生—药理学]