孕酮诱导猪中性粒细胞胞外诱捕网形成及其机制

The functional study of progesterone on the formation of neutrophil extracellular trap in swine

中性粒细胞(polymorphonuclear neutrophils,PMN)胞外诱捕网(neutrophil extracellular traps,NETs)是一种重要的先天性免疫反应机制。为了探究孕酮是否与猪PMN NETs形成相关,本研究通过分离母猪外周血获得PMN,与不同质量浓度孕酮(10,20,40μg/L)共孵育2 h后,通过激光共聚焦显微镜观察NETs的形成;运用picoGreen~?试剂盒定量分析NETs的释放水平;通过乳酸脱氢酶(lactate dehydrogenase,LDH)试剂盒分析细胞外LDH活性。结果表明,孕酮可以显著诱导以DNA为骨架,装饰有PMN弹性蛋白(neutrophil elastase,NE)和髓过氧化物酶(myeloperoxidase,MPO)等蛋白颗粒的NETs形成,NETs形成与孕酮诱导剂量以及孵育时间呈正相关,且孕酮诱导猪PMN释放NETs的过程中,细胞外LDH活性无改变。本研究为有关雌性生殖激素参与调节母猪免疫系统的相关研究提供了新思路。

Neutrophil extracellular traps(NETs)are an important innate immune mechanism,which was discovered in recent years.In this study,to explore whether progesterone can induce the formation of NETs in swine,we used progesterone(10,20,40μg/L)to incubated with neutrophils for 2 h.The formation of NETs induced by progesterone was observed by laser confocal microscopy and quantitatively detected by picoGreen~?.Moreover,the levels of lactate dehydrogenase(LDH)were measured by LDH cytotoxicity assay kit.The results indicated that progesterone significantly induced the formation of NETs,which was a DNA-based network structure modified with neutrophil elastase(NE)and myeloperoxidase(MPO).Futhermore,progesterone-induced NETs was a dose-and time-dependent process,but do not alter extracellular LDH activity.This study provides new clues and foundations for the interaction between sex hormones and innate immunity.