中性粒细胞胞外陷阱（NET）是引发和持续炎症的重要因素。然而，NETs 在幼年特发性关节炎 (JIA) 不同亚型中的作用却很少被研究。因此，我们的目的是探讨 JIA 衍生的中性粒细胞释放 NET 的能力以及 TNF-α（肿瘤坏死因子-α）抑制剂对体外和体内 NET 形成的影响，并评估 NET 衍生产品的关联通过体外刺激和抑制研究评估中性粒细胞释放 NET 的能力以及阿达木单抗对 NET 形成的影响。检测血浆 NET 衍生产物以评估体内 NET 形成的发生率。此外，使用流式细胞术和蛋白质印迹法检测中性粒细胞中与NET相关的信号成分。与来自HC的中性粒细胞相比，来自少关节型JIA、多关节型JIA和附着点炎相关关节炎患者的中性粒细胞更容易自发产生NET。以及体外对 TNF-α 或 PMA 的反应。 JIA患者的外周循环中存在过多的NET形成，血浆中NET衍生产物（游离DNA和MPO-DNA复合物）水平升高可以准确区分JIA患者和HC，并与疾病活动度呈正相关。多元线性回归分析显示，红细胞沉降率和TNF-α水平是自变量，且与游离DNA浓度呈正相关。值得注意的是，TNF-α抑制剂可以在体外和体内有效阻止NET的形成。此外，JIA 来源的中性粒细胞中 NET 相关激酶的磷酸化水平显着升高。我们的数据表明，NET 可能发挥致病作用，并可能参与 JIA 中 TNF-α 介导的炎症。循环中的 NET 衍生产品具有潜在的诊断和疾病监测价值。此外，与JIA患者NET形成分子机制相关的初步结果为NET靶向治疗提供了理论基础。版权所有©2024 Tang，Zhong，Wu，Huang，Liu，Chen，Xi，Wen，He，Yang，Liu 、熊和金。

Neutrophil extracellular traps (NETs) are important factors in initiating and perpetuating inflammation. However, the role of NETs in different subtypes of juvenile idiopathic arthritis (JIA) has been rarely studied. Therefore, we aimed to explore the ability of JIA-derived neutrophils to release NETs and the effect of TNF-α (tumor necrosis factor-alpha) inhibitors on NET formation both in vitro and in vivo, and evaluate the associations of NET-derived products with clinical and immune-related parameters.The ability of neutrophils to release NETs and the effect of adalimumab on NET formation was assessed via in vitro stimulation and inhibition studies. Plasma NET-derived products were detected to assess the incidence of NET formation in vivo. Furthermore, flow cytometry and western blotting were used to detect NET-associated signaling components in neutrophils.Compared to those derived from HCs, neutrophils derived from patients with oligoarticular-JIA, polyarticular-JIA and enthesitis-related arthritis were more prone to generate NETs spontaneously and in response to TNF-α or PMA in vitro. Excessive NET formation existed in peripheral circulation of JIA patients, and elevated plasma levels of NET-derived products (cell-free DNA and MPO-DNA complexes) could accurately distinguish JIA patients from HCs and were positively correlated with disease activity. Multiple linear regression analysis showed that erythrocyte sedimentation rate and TNF-α levels were independent variables and were positively correlated with cell-free DNA concentration. Notably, TNF-α inhibitors could effectively prevent NET formation both in vitro and in vivo. Moreover, the phosphorylation levels of NET-associated kinases in JIA-derived neutrophils were markedly increased.Our data suggest that NETs might play pathogenic roles and may be involved in TNF-α-mediated inflammation in JIA. Circulating NET-derived products possess potential diagnostic and disease monitoring value. Furthermore, the preliminary results related to the molecular mechanisms of NET formation in JIA patients provide a theoretical basis for NET-targeted therapy.Copyright © 2024 Tang, Zhong, Wu, Huang, Liu, Chen, Xi, Wen, He, Yang, Liu, Xiong and Jin.