布鲁菌干扰MФ泛素依赖型自噬通路的研究  

Study on Brucella interfering the ubiquitin-dependent autophagic pathway in macrophage

作　　者：陈冬志[1] 洪洋[1] 李春晓[1] 李娜[2] 曹志然[1] 王家鑫[2] 孟明[1] 

机构地区：[1]河北大学医学部,保定071002 [2]河北农业大学动物科技学院

出　　处：《中华微生物学和免疫学杂志》2012年第9期798-802,共5页Chinese Journal of Microbiology and Immunology

基　　金：河北省自然科学基金项目（C2010000243）;国家“十一五”科技支撑计划项目（2006BAD04A10）

摘　　要：目的探讨布鲁菌减毒株S1330对于小鼠巨噬细胞（MФ）泛素依赖型自噬通路的影响。方法应用布鲁菌S1330株作用于小鼠MФ构建体外感染模型。观察M中内的吞噬过程、泛素化水平及自噬水平。实验设立巨噬细胞正常组、感染组、自噬诱导对照组、自噬诱导感染组，应用吉姆萨染色法、免疫荧光法、Westernblot法分别观察不同时间点、不同组MФ内泛素化蛋白含量及自噬水平变化。结果布鲁菌感染MФ0．5h胞内出现泛素化菌体蛋白，随着感染时间的延长，胞内泛素化蛋白聚集持续增多，12hMФ死亡；与之相对应的自噬体标记物LC3B蛋白表达严重不足，而自噬诱导的感染组MФ内泛素化的菌体蛋白比例减少。结论布鲁菌S1330株感染MФ启动胞内泛素化机制，却干扰泛素依赖型自噬通路的成熟，使泛素化菌体蛋白大量聚集不能有效清除，最终导致了MФ功能障碍而死亡。Objective To investigate the effect on ubiquitin-dependent autophagic pathway in macrophage（MФ） infected by B. suis S1330 attenuated strains. Methods Infected MФ in vitro using Bru- cella S1330 strains to construct experimental model. Observed the process of phagocytic, the level of ubiquit- ination and autophagy in MФ of mice. MФ was divided into control group, infected group, positive control group and infected group after RAPA induced autophagy. The Giemsa staining immunofluorescence and Western blot were applied to observe the chances of ubiquitinated and autophagic protein in MФ at different time points within different groups. Results Ubiquitinated bacterial protein was detected at 0.5 h after in- fected MФ. With the time passing, the ubiquitinated bacterial protein increased and aggregated intracellular until MФ dead at 12 h after infected. The expression of LC3B protein was serious deficiency in MФ which infected group, but ubiquitinated bacterial protein decreased significantly in MФ after RAPA induced. Con- clusion Brucella S1330 stain can arouse intracellular ubiquitination process in infected MФ, and interfere the ubiquitin-dependent autophagic pathway. A large number of aggregated and ubiquitinated bacterial pro- tein can not be effectively removed, it leads to MФ dysfunction and dead.

关 键 词：巨噬细胞 布鲁菌 泛素化 自噬 

分 类 号：R378[医药卫生—病原生物学]