Alamar blue法检测药物细胞内抗结核活性的研究  

Study of Alamar blue assay for determining the intracellular antitubercular activity of new drugs

作　　者：郑梅琴[1] 陆宇[1] 王彬[1] 付雷[1] 

机构地区：[1]北京市结核病胸部肿瘤研究所,北京101149

出　　处：《中国抗生素杂志》2011年第6期468-473,共6页Chinese Journal of Antibiotics

基　　金："重大新药创制"科技重大专项(No.2009ZX09303-005)

摘　　要：目的建立基于Alamar blue法检测的巨噬细胞模型,应用于氯苯吩嗪衍生物细胞内抗结核活性的测定。方法以不同菌量/细胞比例感染巨噬细胞不同时间,阳性药INH、RFP、EMB与感染细胞作用不同时间,CFU计数测定细胞内菌量,考察最适感染复数、感染时间与药物作用时间。以比色法测定一些常用抗生素对感染细胞内菌量的影响,并与CFU计数结果比较,考察比色法的准确性、特异性,并应用比色法测定氯苯吩嗪衍生物的细胞内抗结核活性。结果最适感染复数为10:1,感染4h为宜,药物作用时间为3d。比色法与CFU计数测定药物细胞内抗菌活性的结果一致。所测的吩嗪类化合物显示出良好的细胞内抗菌活性。结论建立了Alamar blue比色法巨噬细胞筛选模型,可对细胞内药物抗结核活性作出快速、客观的评价。Objective To establish an Alamar blue colorimetric macrophage model and apply it to determine the intracellular antitubercular activities of clofazimine derivatives. Methods The optimal MOI, phagocytosed time and drug exposured time were selected based on the bacteria amount in treated macrophages, which was determined by CFU count. The activities of some commonly used antibiotics were investigated within infected macrophages by the colorimetric method and compared with the results obtained from CFU count. The intracellular activities of clofazimine derivatives were determined by the colorimetric method. Results The optimal MOI, phagocytosed time and drug exposured time were 10：1, 4h and 3d respectively. The intracellular activities of the commonly used antibiotics obtained from the colorimetric method were approximate to that from CFU count. The tested clofazimine derivatives showed good intracellular antitubercular activities. Conclusion An Alamar blue colorimetric macrophage model was established and enabled to objectively evaluate the intracellular antitubercular activities of drugs within a short period of time.

关 键 词：药物筛选 巨噬细胞 Alamar BLUE 

分 类 号：R978.3[医药卫生—药品]