天津医药

• 细胞与分子生物学 • 上一篇    下一篇

Syk 参与单增李斯特菌感染中炎症复合体的形成

刘倩倩,申艳娜,刘运德   

  1. 天津医科大学医学检验学院
  • 收稿日期:2014-01-14 修回日期:2014-03-01 出版日期:2014-05-15 发布日期:2014-05-15
  • 通讯作者: 刘倩倩

Syk kinase Signaling is Required for the Inflammasomes Activation during Listeria monocytogenes Infection

  • Received:2014-01-14 Revised:2014-03-01 Published:2014-05-15 Online:2014-05-15
  • Contact: Qian-Qian LIU

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摘要： 【摘要】 目的 本研究旨在探讨单增李斯特菌（Listeria monocytogenes, LM）感染过程中炎症复合体形成的分子机制。方法 LM分别感染syk，P38MAPK以及PI3K抑制剂组小鼠腹腔由来巨噬细胞后，利用ELISA方法检测上清液中IL-18的蛋白水平；免疫荧光观察细胞内炎症复合体的关键蛋白组分ASC的变化情况；western blot检测LM感染巨噬细胞不同时间点syk蛋白激酶的磷酸化水平。结果 LM感染后，syk抑制剂组产生的IL-18蛋白水平与未处理组相比显著降低，而其他抑制剂组产生的IL-18的蛋白水平未明显改变，进而免疫荧光结果又显示syk抑制剂可明显降低胞内ASC-speck的形成个数。syk磷酸化水平在LM感染5min即可被检测到，进一步证实syk参与炎症复合体活化的过程。结论 syk参与调控单增李斯特菌感染过程中炎症复合体的活化。

关键词: syk, 单核细胞增生李斯特菌, 炎症小体

Abstract: [Abstract] Objective The aim of this study is to clarify the role of syk kinase in inflammasomes activation during Listeria monocytogenes (LM) infection. Methods Murine peritoneal exudates macrophages pretreated with inhibitor of syk, P38MAPK, or PI3K kinase were infected with LM for 24h, the protein level of IL-18 in the supernatant was detected by ELISA kit, the activation condition of key molecule ASC in the infected-macrophages cytoplasm was observed under fluorescence microscope. Furthermore, the syk protein kinase phosphorylation level at different time point during LM infection were detected by western blot. Results IL-18 protein level produced by macrophages in the presence of syk inhibitor was significantly reduced compared with that in macrophages without pretreatment. Meanwhile, the percentage of ASC-speck formation in the cytoplasm of syk inhibitor pretreated-- macrophages infected with LM diminished apparently. Furthermore, we found that the phosphorylation level of syk could be detectable at 5min postinfection, which confirmed the critical role of syk in the inflammasomes activation upon LM infection. Conclusion Syk kinase signaling is required for the inflammasomes activation upon Listeria monocytogenes infection in murine macrophages.

Key words: syk, listeria monocytogenes, inflammasome