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半滑舌鳎(Cynoglossus semilaevis)中性粒细胞胞外陷阱(neutrophil extracellular traps):产生特征以及抗菌作用
Thesis Advisor孙黎
Degree Grantor中国科学院大学
Place of Conferral北京
Degree Discipline海洋生物学
Keyword中性粒细胞胞外陷阱 Netosis 半滑舌鳎 抗菌效果 天然免疫防御
中性粒细胞胞外陷阱(NETs)是中性粒细胞释放到胞外的网状纤维结构,它作为机体一种细胞免疫防御机制抵抗了微生物的入侵。NETs是以DNA作为基本结构骨架,其上间隙附着大量的蛋白结构。NETs的产生被称作“NETosis”。NETosis具有两种方式:一种是自杀性的,即细胞在释放NETs的过程中死亡,另一种是在释放NETs的过程中仍然保持细胞活性。目前,关于哺乳动物中的NETosis研究很多,但是对鱼类中的NETosis却知之甚少。半滑舌鳎(Cynoglossus semilaevis)是一种在中国具有非常重要经济价值的硬骨鱼;在本研究中,我们研究了半滑舌鳎中的NETosis。实验证明,在佛波酯(phorbol 12-myristate 13-acetate,PMA)和三种不同的鱼类致病菌荧光假单胞菌(Pseudomonas fluorescens)、哈维氏弧菌(Vibrio harveyi)和迟缓爱德华氏菌(Edwardsiella tarda)刺激下,中性粒细胞能够在活性状态下释放大量的NETs结构。
半滑舌鳎中性粒细胞释放的NETs能够捕获细菌,但却不能杀死细菌。胞内复制实验表明,NETs能够抑制胞外菌(荧光假单胞菌和哈维氏弧菌)的繁殖,但不能抑制胞内菌(迟缓爱德华氏菌)的繁殖。在中性粒细胞产生NETs的过程中,活性氧(reactive oxygen species,ROS)、一氧化氮(nitric oxide,NO)和髓过氧化物酶(myeloperoxidase,MPO)的产生量也有显著增加,并且,用相应的抑制剂抑制ROS、NO和MPO的产生显著降低了PMA以及细菌诱导的NETs量。总的来说,我们的研究结果第一次阐明了在硬骨鱼类中存在着不依赖细胞死亡的NETosis,半滑舌鳎的中性粒细胞能够产生具有抗菌效果的NETs,并且其NETs的产生需要有ROS、NO和MPO的存在。总的来说,这些新发现有助于进一步理解鱼类NETosis的机制以及NETs在鱼类中的功能。
Other AbstractNeutrophil extracellular traps (NETs) are structures released by neutrophils as a cellular immune defense against microbial invasion. The process of NETs generation, netosis (NETosis), can take place via either a suicidal mechanism, during which the NETs releasing cells became dead, or a “live” mechanism, during which the NETs-releasing cells remain vital. NETosis has been studied intensively in mammals in recent years, but very little is known about the NETosis in fsh. In this study, we examined NETosis in tongue sole (Cynoglossus semilaevis), a species of teleost with important economic values. We found that following stimulation with phorbol 12-myristate 13-acetate (PMA) and three common fsh bacterial pathogens, abundant NETs structures were released by neutrophils that were most likely in a live state.
 The released NETs captured, but did not kill, the bacterial pathogens; however, the replication of extracellular, but not intracellular, pathogens was inhibited by NETs to significant extents. Reactive oxygen species (ROS), nitric oxide (NO), and myeloperoxidase (MPO) production were observedto be enhanced in NETosing neutrophils, and blocking the production of these factors by inhibitors signifcantly decreased NETs production induced by PMA and all three bacteria. Taken together, these results indicate for the frst time that in teleost there exists a non-cell death pathway of NETosis that produces NETs with antibacterial effects in a ROS-, NO-, and MPO-dependent manner. Taken together, these findings add new insights into the mechanism of NETosis and the functionality of NETs in fish.
Subject Area生物学 ; 细胞免疫学
Document Type学位论文
Recommended Citation
GB/T 7714
赵明丽. 半滑舌鳎(Cynoglossus semilaevis)中性粒细胞胞外陷阱(neutrophil extracellular traps):产生特征以及抗菌作用[D]. 北京. 中国科学院大学,2017.
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