生殖支原体脂质相关膜蛋白经Toll样受体2激活NF-κB  

Lipid-associated membrane proteins of Mycoplasma genitalium activate NF-κB via Toll-like receptors 2

作　　者：何军[1] 游晓星[2] 曾焱华[2] 伍宁[1] 吴移谋[2] 

机构地区：[1]南华大学附属南华医院,衡阳421000 [2]南华大学病原生物学研究所,衡阳421001

出　　处：《中华微生物学和免疫学杂志》2010年第12期1137-1140,共4页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目（30770115）;湖南省教育厅资助项目（10C1133）

摘　　要：目的 探讨生殖支原体（Mg）脂质相关膜蛋白（LAMPs）能否经Toll样受体2（TLR2）激活核转录因子κB（NF-κB）.方法 提取MgLAMPs刺激THP-1细胞,ELISA检测NF-κBp65活化水平,RT-PCR检测THP-1细胞TLR2 mRNA的表达,随后ELISA检测TLR2中和抗体对LAMPs激活NF-κBp65的影响,最后用LAMPs刺激瞬时共转染pFLAG-TLR2、pNF-κB-luc、pRL-TK的293T细胞,双荧光素酶报告基因分析TLR2在LAMPs激活NF-κB中的作用.结果 LAMPs能诱导THP-1细胞激活NF-κBp65,并且与LAMPs浓度呈明显的剂量依赖性,当LAMPs为4.0μg/ml时NF-κBp65活化程度最高;LAMPs能上调THP-1细胞TLR2 mRNA的表达;TLR2中和抗体能使LAMPs激活NF-κBp65的活化程度降低60%;共转染pFLAG-TLR2浓度的增加,NF-κB的活化程度明显增加,并且与TLR2的转染量呈剂量依赖性.结论 Mg LAMPs能经TLR2激活NF-κB,TLR2介导的激活在LAMPs发挥致病性过程起着重要作用.Objective To investigate whether nuclear transcription factor κB（NF-κB） through Toll-like receptors 2（TLR2） was activated by lipid-associated membrane proteins（LAMPs） of Mycoplasma genitalium.Methods LAMPs were extractded and THP-1 cells were stimulated.The activation of NF-κBp65 was detected by ELISA and the expression of TLR2 mRNA was detected by RT-PCR.Effects of TLR2 neutralizing antibody on LAMPs induced the activation of NF-κBp65 was analyzed by ELISA.After LAMPs stimulated 293T cells with the co-transfection pFLAG-TLR2,pNF-κB-luc,pRL-TK,the activity of NF-κB firefly luciferase and pRL-TK Renilla luciferase were detected by the dual-luciferase reporter gene,to analyzed the role of TLR2-mediated NF-κB activation by LAMPs in 293T cells.Results The activation of NF-κBp65 was mediated in LAMPs induced THP-1 cells and was significantly increased by LAMPs in a dose dependent manner.when LAMPs was 4.0 μg/ml,the activation of NF-κBp65 was the highest level.TLR2 mRNA expression was up-regulated by LAMPs in THP-1 cells.TLR2 neutralizing antibody could inhibit the activation of NF-κB by 60% in LAMPs stimulated THP-1.NF-κB fluorescence was significantly increased by co-transfection pFLAG-TLR2 in a dose-dependent manner. ConclusionMycoplasma genitalium-derived lipid-associated membrane proteins activate NF-κB via TLR2 and the activation of TLR2-mediated play an important role in pathogenic process of LAMPs.

关 键 词：生殖支原体 脂质相关膜蛋白 TOLL样受体2 核转录因子ΚB 

分 类 号：R759[医药卫生—皮肤病学与性病学]