| 吴 华，卿文衡，刘 军，张黎黎.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.[J].中南医学科学杂志.,2015,43(6):614-617. |
| 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 |
| 投稿时间：2015-09-10 |
| DOI： |
| 中文关键词: 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 |
| 英文关键词:Chlamydophila pneumonia Toll-like receptor 2 TLR4 TNF-α IL-8 |
| 基金项目:湖南省卫生厅科技计划项目（B2013-132）. |
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| 中文摘要: |
| 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50.0 μg/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。 |
| 英文摘要: |
| Objective To investigate the molecular mechanism of the inclusion membrane protein Cpn0147 on the secretion of cytokines in monocytes. Methods Cultured THP-1 cells were stimulated with different concentration of endotoxin-free recombinant inclusion protein （1.0,10.0,30.0 and 50.0 μg/mL） for 0～48 h.Secretion of TNF-α and IL-8 were detected 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-α and IL-8 were detected by ELISA. Results Cpn0147 could induce THP-1 cells the secretion of TNF-α and IL-8,and the expression of mRNA.The production of TNF-α and 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 antibody significantly abrogate Cpn0147-induced cytokines production,similar results was also obtained by RNA interference of TLR2 and TLR4. Conclusion Cpn0147 can induce TNF-α and IL-8 secretion via TLR2 and TLR4. |
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