NMDA receptor modulation of glutamate release in activated neutrophils
NMDA receptor modulation of glutamate release in activated neutrophils
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DOI：
10.1016/j.ebiom.2019.08.004
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发表时间：
2019-09-01
期刊：
影响因子：
11.1
通讯作者：
Ackland, Gareth L.
中科院分区：
文献类型：
--
作者：
del Arroyo, Ana Gutierrez;Hadjihambi, Anna;Ackland, Gareth L.
背景：神经元耗竭改善实验性脓毒症/脑损伤的神经功能结局。我们假设，中性粒细胞可能会加剧神经元损伤，通过释放的神经毒性量的神经递质glutamate.Methods：实时谷氨酸释放的主要人类中性粒细胞使用酶生物传感器。在存在/不存在N-甲基-D-天冬氨酸受体（NMDAR）拮抗剂的情况下，比较了人全血、分离的中性粒细胞或人细胞系中中性粒细胞的细菌和直接蛋白激酶C（佛波醇12-肉豆蔻酸酯13-乙酸酯; PMA）活化。使用流式细胞术（吞噬作用，活性氧（ROS）的产生）和实时呼吸测定法（耗氧量）比较了NMDAR支架蛋白缺乏的野生型和转基因小鼠中性粒细胞的中性粒细胞的细菌和直接激活：谷氨酸和NMDAR共激动剂D-丝氨酸迅速释放的中性粒细胞响应细菌和PMA诱导的激活。药理学NMDAR阻断减少谷氨酸、D-丝氨酸的自分泌释放和活化的原代人中性粒细胞的呼吸爆发。限制NMDAR介导的神经元损伤的高度特异性小分子抑制剂ZL 006也通过腹膜炎小鼠模型中活化的中性粒细胞减少ROS，这是通过NMDAR GluN 2B亚基与其支架蛋白突触后密度蛋白-95（PSD-95）解偶联实现的。PSD-95的基因切除减少了活化的鼠嗜中性粒细胞的ROS产生。NMDAR GluN 2B亚基的药理学阻断减少了荧光假单胞菌诱导的原发性人中性粒细胞活化，荧光假单胞菌是一种分泌谷氨酸的革兰氏阴性杆菌，与引起医院获得性infections.Interpretation的病原体密切相关：这些数据表明，激活的中性粒细胞释放谷氨酸通过对这些细胞表达的NMDAR的作用以自分泌方式增强ROS的产生。(C)2019作者（S）由爱思唯尔公司出版
Background: Neutrophil depletion improves neurologic outcomes in experimental sepsis/brain injury. We hypothesized that neutrophils may exacerbate neuronal injury through the release of neurotoxic quantities of the neurotransmitter glutamate.Methods: Real-time glutamate release by primary human neutrophils was determined using enzymatic biosensors. Bacterial and direct protein-kinase C (Phorbol 12-myristate 13-acetate; PMA) activation of neutrophils in human whole blood, isolated neutrophils or human cell lines were compared in the presence/absence of N-Methyl-D-aspartic acid receptor (NMDAR) antagonists. Bacterial and direct activation of neutrophils from wildtype and transgenic murine neutrophils deficient in NMDAR-scaffolding proteins were compared using flow cytometry (phagocytosis, reactive oxygen species (ROS) generation) and real-time respirometry (oxygen consumption).Findings: Both glutamate and the NMDAR co-agonist D-serine are rapidly released by neutrophils in response to bacterial and PMA-induced activation. Pharmacological NMDAR blockade reduced both the autocrine release of glutamate, D-serine and the respiratory burst by activated primary human neutrophils. A highly specific small-molecule inhibitor ZL006 that limits NMDAR-mediated neuronal injury also reduced ROS by activated neutrophils in a murine model of peritonitis, via uncoupling of the NMDAR GluN2B subunit from its' scaffolding protein, postsynaptic density protein-95 (PSD-95). Genetic ablation of PSD-95 reduced ROS production by activated murine neutrophils. Pharmacological blockade of the NMDAR GluN2B subunit reduced primary human neutrophil activation induced by Pseudomonas fluorescens, a glutamate-secreting Gram-negative bacillus closely related to pathogens that cause hospital-acquired infections.Interpretation: These data suggest that release of glutamate by activated neutrophils augments ROS production in an autocrine manner via actions on NMDAR expressed by these cells. (C) 2019 The Author(s). Published by Elsevier B.V.