A族链球菌感染巨噬细胞激活TLR2及TLR4表达的研究  

Study on the activation of TLR2 and TLR4 in group A streptococcus-infected macrophages

作　　者：王海欣 陈继科 郭宜 时建华 吴景华[3] 

机构地区：[1]唐山市中医医院口腔科,唐山063000 [2]唐山市协和医院口腔科 [3]河北联合大学附属医院

出　　处：《山西医科大学学报》2015年第1期21-24,100,共5页Journal of Shanxi Medical University

基　　金：唐山市科技局指导项目(14130262a)

摘　　要：目的通过分析A族链球菌(GAS)感染巨噬细胞后TLR2及TLR4的活化情况,以此来研究GAS感染后产生炎症反应的信号通路,为GAS感染巨噬细胞的预防及治疗奠定基础。方法 GAS及热灭活GAS以感染复数(MOI)为100∶1感染小鼠巨噬细胞RAW264.7细胞后1,2,4,6 h,提取总RNA采用real-time PCR技术检测TLR2/4的表达情况;同时GAS及热灭活的GAS注射C57小鼠腹腔,24 h后分离腹腔巨噬细胞并提取蛋白,利用Western blot及免疫荧光技术对TLR2和TLR4的表达量进行分析。结果巨噬细胞表面TLR2及TLR4在GAS感染后1 h活化不明显,但作用2 h后TLR2及TLR4表达明显升高(P<0.05),4 h时达到高峰,6 h后开始降低。热灭活GAS感染巨噬细胞TLR2的表达与GAS感染比较未见到统计学差异(P>0.05),但巨噬细胞表面TLR4的表达与GAS组具有明显的差异(P<0.05)。结论 GAS感染小鼠巨噬细胞可同时激活TLR2及TLR4受体,其中TLR4受体主要通过位于GAS表面及其分泌的活性蛋白成分激活,TLR2受体则主要由其菌体成分进行活化。Objective To investigate the expression of TLR2 and TLR4 in group A streplLococcus（GAS）-infected macrophages and to explore the pathway in GAS-induced inflammation for providing a potential new way in prevention and treatment of GAS-induced infection. Methods The macrophages were infected by GAS and heat-inactivated GAS at MOI of 100： 1. After infection for 1,2, 4, 6 h, the total RNA was separated and the expression of TLR2 or TLR4 was identified by real-time PCR. The peritoneal macrophages infected by GAS or heat-inactivated GAS for 24 h were separated and the activation of TLR2 or TLR4 was analyzed by immunofluorescence and Western blot. Results TLR2 and TLR4 expression levels showed no obvious changes in GAS-infected macrophages at 1 h, and significantly increased at 2 h（P 〈 0.05 ）, peaked at 4 h and decreased at 6 h. There was significant difference in TLR4 expression between heat-inactivated GAS-infected cells and GAS-infected cells （ P 〈 0.05 ）, but not in TLR2 expression （ P 〉 O. 05 ）. Conclusion TLR2 and TLR4 could be simultaneously activated in GAS-infected macrophages. TLR4 raay be activated by the membrane proteins or secreted proteins of GAS, while TLR2 activation may be induced by GAS itself.

关 键 词：GAS 巨噬细胞 TLR2 TLR4 

分 类 号：R392.12[医药卫生—免疫学]