肺炎嗜衣原体包涵体膜蛋白Cpn0147诱导单核细胞分泌TNF-α和IL-8  

Chlamydophila pneumonia-Derived Inclusion Membrane Protein Cpn0147 Induces Monocytes Secretion of TNF-α and IL-8

作　　者：吴华 卿文衡 刘军 张黎黎 

机构地区：[1]衡阳市中心医院检验科,湖南衡阳421001

出　　处：《中南医学科学杂志》2015年第6期614-617,684,共5页Medical Science Journal of Central South China

基　　金：湖南省卫生厅科技计划项目(B2013-132)

摘　　要：目的：探讨肺炎嗜衣原体（ Cpn）包涵体膜蛋白Cpn0147诱导人单核细胞分泌炎症因子的分子机制。方法用去除内毒素活性的不同浓度Cpn0147重组蛋白（1.0、10.0、30.0和50.0μg/mL）刺激THP-1细胞0~48 h,ELISA检测肿瘤坏死因子-α（TNF-α）和白介素-8（IL-8）的分泌水平；实时定量PCR检测TNF-α和IL-8 mRNA的表达；用Toll样受体2（ TLR2）和TLR4中和抗体处理THP-1细胞,或采用 TLR2和 TLR4 siRNA沉默其表达, ELISA检测THP-1处理前后TNF-α和IL-8分泌的变化。结果 Cpn0147可诱导THP-1细胞表达TNF-α和IL-8的mRNA和蛋白；同时,不同时间点Cpn0147对TNF-α和IL-8的诱导水平有所不同,Cpn0147作用2~6 h后即可诱导TNF-α和IL-8分泌,12 h时达到峰值,随后逐渐下降。采用TLR2和TLR4中和抗体封闭THP-1细胞后,TNF-α和IL-8分泌水平明显减少,采用TLR2和TLR4 siRNA干扰其表达后,TNF-α和IL-8的分泌也得到了类似的结果。结论 Cpn0147可能通过TLR2和TLR4介导TNF-α和IL-8分泌。Objective To investigate the molecular mechanism of the inclusion membrane protein Cpn0147 on the se-cretion of cytokines in monocytes. Methods Cultured THP-1 cells were stimulated with different concentration of endotox-in-free recombinant inclusion protein （1. 0,10. 0,30. 0 and 50. 0μg/mL） for 0~48 h. Secretion of TNF-αand IL-8 were de-tected by ELISA,expression of the mRNA was measured by real-time PCR. In addition,cells were preincubated with Toll-like receptor 2（TLR2） or TLR4 neutralizing antibody,or transfected with siRNA for TLR2 and TLR4,secretion of TNF-αand IL-8 were detected by ELISA. Results Cpn0147 could induce THP-1 cells the secretion of TNF-αand IL-8,and the expres-sion of mRNA. The production of TNF-αand IL-8 varies with time intervals,the ELISA results indicated secretion of cytokines appeared after 2~6 h of stimulation,peaked at 12 h and then decreased. Pre-incubation of TLR2 or TLR4 neutralizing anti-body significantly abrogate Cpn0147-induced cytokines production,similar results was also obtained by RNA interference of TLR2 and TLR4. Conclusion Cpn0147 can induce TNF-αand IL-8 secretion via TLR2 and TLR4.

关 键 词：肺炎嗜衣原体 TOLL样受体2 Toll样受体4 肿瘤坏死因子-α 白介素-8 

分 类 号：R392.12[医药卫生—免疫学]