不同毒力结核分支杆菌感染RAW264.7巨噬细胞不同时间细胞因子转录水平标准曲线的建立及应用  

Establishment and Application a Standard Curve of Quantitative Real- time PCR for Cytokine Transcript Levels after Infected by Different Virulence Mycobacterium tuberculosis at Different Time Point

作　　者：张妍[1] 宋纪伟[2] 曾范利[3] 时坤[3] 李健明[3] 刘杨[1] 刘菲[1] 孙凡婷 杜锐[3,4] 

机构地区：[1]吉林农业大学动物科学技术学院,长春130118 [2]吉林市人民医院,吉林吉林132001 [3]吉林农业大学中药材学院,长春130118 [4]教育部动物生产及产品质量安全重点实验室,吉林省药用动物二级实验室,长春130118

出　　处：《中国兽药杂志》2014年第12期5-10,共6页Chinese Journal of Veterinary Drug

基　　金：国家科技支撑计划(2011BAI03B02-1);吉林省科技厅科技支撑计划(20120225)

摘　　要：为检测不同毒力结核分枝杆菌感染巨噬细胞后细胞因子转录水平的变化,构建IL-6、IL-10、TNF-α重组标准质粒,建立了实时荧光定量PCR标准曲线。应用该方法对不同毒力结核分枝杆菌感染RAW264.7巨噬细胞6个时间点细胞因子的转录水平进行分析,结果显示H37Rv组、BCG组相比于对照组刺激后均使三种细胞因子的转录水平发生变化,其中IL-6和TNF-α发生明显变化。本试验通过对不同时间点细胞因子转录水平的分析为结核分枝杆菌对巨噬细胞凋亡机制的研究提供新思路。In order to detect the change of cytokine transcript levels after infected by Mycobacterium tuberculosis, build recombinant standard plasmids of IL-6, IL-10,TNF-α, a quantitative real-time PCR standard curve was established for cytokine transcript levels. Application of the method was used to analyze cytokine transcript levels that different virulence Mycobacterium tuberculosis infecting RAW264. 7 macrophages at six time points. Comparison with the control group, H37Rv and BCG made changes in transcript levels of these three cytokines, while the IL-6 and TNF-α have more obvious varies. This study provided new ideas of mechanism between Mycobacterium tuberculosis and macrophages through different time points cytokines transcriptional level.

关 键 词：结核分枝杆菌 RAW264.7细胞 荧光定量PCR 细胞因子 

分 类 号：S852.618[农业科学—基础兽医学]