IL-8 通过烟酰胺腺嘌呤二核苷酸磷酸氧化酶和丝裂原活化蛋白激酶途径依赖性机制触发葡萄膜炎中性粒细胞细胞外陷阱的形成。
IL-8 Triggers Neutrophil Extracellular Trap Formation Through an Nicotinamide Adenine Dinucleotide Phosphate Oxidase- and Mitogen-Activated Protein Kinase Pathway-Dependent Mechanism in Uveitis.
机构信息
The First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Ophthalmology, Chongqing Eye Institute, Chongqing Branch (Municipality Division) of National Clinical Research Center for Ocular Diseases, Chongqing, China.
Department of Hepatobiliary Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
出版信息
Invest Ophthalmol Vis Sci. 2023 Oct 3;64(13):19. doi: 10.1167/iovs.64.13.19.
PURPOSE
To explore the mechanism underlying IL-8-induced neutrophil extracellular trap (NET) formation in patients with ocular-active Behçet's disease (BD) and the effect of inhibiting NET formation on the severity of inflammation in experimental autoimmune uveitis (EAU) mice.
METHODS
The serum extracellular DNA and neutrophil elastase (NE) and IL-8 levels in patients with ocular-active BD, the expression of myeloperoxidase, NE, and histone H3Cit in IL-8-induced neutrophils isolated from healthy controls, and the effects of NETs on HMC3 cells were detected. Female C57BL/6J mice were immunized with IRBP651-670 to induce EAU and EAU mice received intravitreal injection of the CXCR2 (IL-8 receptor) antagonist SB225002 or PBS. The serum levels of extracellular DNA, NE, and keratinocyte-derived chemokine (the mouse ortholog of human IL-8) and expression of myeloperoxidase, NE, and histone H3Cit in mouse retinas were detected. Disease severity was evaluated by clinical and histopathological scores.
RESULTS
Serum keratinocyte-derived chemokine expression levels in EAU mice and IL-8 expression levels in patients with ocular-active BD increased. IL-8 notably increased NET formation in a dose-dependent manner through an nicotinamide adenine dinucleotide phosphate oxidase and mitogen-activated protein kinase pathway dependent mechanism. IL-8-induced NET formation damaged HMC3 cells in vitro. Pretreatment with SB225002 notably ameliorated the production of NETs in EAU mice.
CONCLUSIONS
Our data confirm that NET formation is induced by IL-8. IL-8-induced NET formation was found to be related to mitogen-activated protein kinase and nicotinamide adenine dinucleotide phosphate pathways. Pretreatment with the CXCR2 antagonist SB225002 alleviated neutrophil infiltration and suppressed NET formation in EAU mice.
目的
探讨白细胞介素-8（IL-8）诱导眼型贝赫切特病（BD）患者中性粒细胞胞外诱捕网（NET）形成的机制，以及抑制 NET 形成对实验性自身免疫性葡萄膜炎（EAU）小鼠炎症严重程度的影响。
方法
检测眼型 BD 患者血清细胞外 DNA 和中性粒细胞弹性蛋白酶（NE）及 IL-8 水平，检测健康对照者 IL-8 诱导的中性粒细胞中髓过氧化物酶、NE 和组蛋白 H3Cit 的表达，检测 NET 对 HMC3 细胞的影响。用 IRBP651-670 免疫雌性 C57BL/6J 小鼠诱导 EAU，EAU 小鼠玻璃体腔注射 CXCR2（IL-8 受体）拮抗剂 SB225002 或 PBS。检测血清细胞外 DNA、NE 和角蛋白细胞来源趋化因子（人 IL-8 的小鼠同源物）水平，检测小鼠视网膜中髓过氧化物酶、NE 和组蛋白 H3Cit 的表达。通过临床和组织病理学评分评估疾病严重程度。
结果
EAU 小鼠血清角蛋白细胞来源趋化因子表达水平和眼型 BD 患者 IL-8 表达水平升高。IL-8 通过烟酰胺腺嘌呤二核苷酸磷酸氧化酶和丝裂原活化蛋白激酶途径依赖性机制显著增加 NET 的形成，呈剂量依赖性。IL-8 诱导的 NET 形成可体外损伤 HMC3 细胞。SB225002 预处理显著改善了 EAU 小鼠 NET 的产生。
结论
本研究数据证实 NET 的形成是由 IL-8 诱导的。发现 IL-8 诱导的 NET 形成与丝裂原活化蛋白激酶和烟酰胺腺嘌呤二核苷酸磷酸途径有关。CXCR2 拮抗剂 SB225002 预处理可减轻 EAU 小鼠中性粒细胞浸润并抑制 NET 形成。
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