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目的探讨铜绿假单胞菌生物被膜对巨噬细胞的免疫逃逸作用以及相关机制。方法用PMA刺激THP-1细胞获得巨噬细胞模型。用6孔板建膜法获得铜绿假单胞菌生物被膜菌。分别用铜绿假单胞菌的生物被膜菌和浮游菌感染巨噬细胞,观察巨噬细胞形态的变化,并检测巨噬细胞的细胞毒性和吞噬功能的变化。进一步用ELISA试剂盒检测感染的巨噬细胞培养上清中炎症细胞因子IL-1β的变化。结果成功构建巨噬细胞模型和铜绿假单胞菌生物被膜菌模型。与感染了浮游菌的细胞相比,感染了生物被膜菌的巨噬细胞形态变化小,释放的LDH降低,吞噬功能减弱,IL-1β的表达量减少。结论铜绿假单胞菌生物被膜菌可以逃逸巨噬细胞的免疫防御作用,其机制可能与降低巨噬细胞的炎症反应有关。
Objective To investigate the immune escape of macrophages from Pseudomonas aeruginosa biofilms and its related mechanism. Methods PMA stimulated THP-1 cells to obtain macrophage model. Pseudomonas aeruginosa biofilm was obtained using a 6-well plate. Macrophages were infected with biofilms and planktonic bacteria of Pseudomonas aeruginosa respectively to observe the morphological changes of macrophages and the changes of macrophages cytotoxicity and phagocytosis. The changes of inflammatory cytokines IL-1β in infected macrophage culture supernatants were further detected by ELISA kit. Results Macrophage model and Pseudomonas aeruginosa biofilm model were successfully constructed. Compared with the cells infected with the planktonic bacteria, macrophages infected with biofilm are less changed in morphology, the released LDH is reduced, the phagocytic function is weakened, and the expression of IL-1β is decreased. Conclusion Pseudomonas aeruginosa biofilm can escape the immune defensive effect of macrophages, and its mechanism may be related to reducing the inflammatory response of macrophages.