Peculiarities of Neutrophil Extracellular Traps Formation in Chinchilla Rabbits

Q3 Agricultural and Biological Sciences Moscow University Biological Sciences Bulletin Pub Date : 2024-08-29 DOI:10.3103/s0096392524600583
N. V. Vorobjeva, M. S. Muntyan
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Abstract

Neutrophil extracellular traps (NETs) are decondensed nuclear chromatin decorated with bactericidal proteins of various cell organelles and performing an effector function aimed at combating pathogens in the inflammatory focus. At the same time, NETs play an important role in the pathogenesis of many autoimmune and inflammatory diseases as well as malignancies. Rabbits are one of the most commonly used species of laboratory animals in medical and biological research. A large number of models of various diseases of the cardiovascular, immune and other human systems have been developed on rabbits. However, in the scientific literature, there is no information about the ability of rabbit neutrophils to undergo NETosis (the programmed cell death in the process of NETs formation) in response to well-known pharmacological stimuli. The purpose of the present study was to investigate in the in vitro system the ability of neutrophils of Soviet chinchilla rabbit to form NETs in response to mimetic of diacylglycerol phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187. To isolate rabbit neutrophils, the Ficoll-Hypaque one-step density gradient centrifugation with modifications was used. Oxidative burst was assessed with luminol-amplified chemiluminescence method, while NETs formation was assessed with immunofluorescence analysis. The study shows for the first time that neutrophils of Soviet chinchilla rabbit do not form NETs in response to PMA but do form traps in response to A23187, and have low levels of oxidative burst in response to PMA, A23187 and chemoattractant N-formyl-methionyl-leucyl-phenylalanine.

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栗鼠兔中性粒细胞胞外陷阱形成的特殊性
摘要中性粒细胞胞外捕获物(NET)是由各种细胞器的杀菌蛋白装饰的解聚核染色质,具有在炎症病灶中对抗病原体的效应功能。同时,NETs 在许多自身免疫性疾病、炎症性疾病以及恶性肿瘤的发病机制中发挥着重要作用。兔子是医学和生物学研究中最常用的实验动物物种之一。以家兔为研究对象开发了大量心血管、免疫和其他人体系统疾病的模型。然而,在科学文献中,还没有关于兔子中性粒细胞在众所周知的药理刺激下发生 NETosis(NETs 形成过程中的程序性细胞死亡)的能力的信息。本研究的目的是在体外系统中研究苏联绒毛兔的中性粒细胞在二酰基甘油的模拟物--光甘油 12-肉豆蔻酸 13-乙酸酯(PMA)和钙离子诱导剂 A23187 的作用下形成 NETs 的能力。为了分离兔中性粒细胞,采用了经过改良的 Ficoll-Hypaque 一步法密度梯度离心法。氧化爆发用发光酚扩增化学发光法进行评估,NETs的形成用免疫荧光分析法进行评估。该研究首次表明,苏联栗鼠兔的中性粒细胞对 PMA 不形成 NET,但对 A23187 有反应,并对 PMA、A23187 和趋化吸引剂 N-甲酰基蛋氨酰亮氨酰苯丙氨酸有低水平的氧化爆发反应。
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来源期刊
Moscow University Biological Sciences Bulletin
Moscow University Biological Sciences Bulletin Agricultural and Biological Sciences-Agricultural and Biological Sciences (all)
CiteScore
1.00
自引率
0.00%
发文量
18
期刊介绍: Moscow University Biological Sciences Bulletin  is forum for research in all important areas of modern biology. It publishes original work on qualitative, analytical and experimental aspects of research. The scope of articles to be considered includes plant biology, zoology, ecology, evolutionary biology, biophysics, genetics, genomics, proteomics, molecular biology, cell biology, biochemistry, endocrinology, immunology, physiology, pharmacology, neuroscience, gerontology, developmental biology, bioinformatics, bioengineering, virology, and microbiology.
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