Laboratorio de Inmunidad Innata, Instituto de Medicina Experimental (IMEX)-CONICET, Academia Nacional de Medicina, Buenos Aires, Argentina.
J Innate Immun. 2017;9(4):387-402. doi: 10.1159/000460293. Epub 2017 May 4.
Tissue injury leads to the release of uric acid (UA). At high local concentrations, UA can form monosodium urate crystals (MSU). MSU and UA stimulate neutrophils to release extracellular traps (NET). Here, we investigated whether these NET could be involved in the development of inflammation by stimulating cytokine release by airway epithelial cells. We found that NET significantly increased the secretion of CXCL8/IL-8 and IL-6 by alveolar and bronchial epithelial cells. These effects were not observed when NETosis was inhibited by Diphenyleneiodonium, elastase inhibitor, or Cl-amidine. Similar findings were made with NET induced by cigarette smoke extract, suggesting that NET proinflammatory capacity is independent of the inducing stimulus. Furthermore, NET affected neither the viability and morphology of epithelial cells nor the barrier integrity of polarized cells. The epithelial stimulatory capacity of NET was not affected by degradation of DNA with micrococcal nuclease, treatment with heparin, or inhibition of the elastase immobilized to DNA, but it was significantly reduced by pretreatment with an anti-HMGB-1 blocking antibody. Altogether, our findings indicate that NET exert direct proinflammatory effects on airway epithelial cells that might contribute in vivo to the further recruitment of neutrophils and the perpetuation of inflammation upon lung tissue damage.
组织损伤导致尿酸(UA)释放。在局部高浓度下,UA 可以形成单钠尿酸盐晶体(MSU)。MSU 和 UA 刺激中性粒细胞释放细胞外陷阱(NET)。在这里,我们研究了这些 NET 是否可以通过刺激气道上皮细胞释放细胞因子来参与炎症的发展。我们发现 NET 显著增加了肺泡和支气管上皮细胞分泌的 CXCL8/IL-8 和 IL-6。当 NETosis 被二苯乙烯碘二酮、弹性蛋白酶抑制剂或 Cl-amidine 抑制时,没有观察到这些作用。用香烟烟雾提取物诱导的 NET 也有类似的发现,表明 NET 的促炎能力与诱导刺激无关。此外,NET 既不影响上皮细胞的活力和形态,也不影响极化细胞的屏障完整性。NET 对上皮细胞的刺激能力不受微球菌核酸酶降解 DNA、肝素处理或固定在 DNA 上的弹性蛋白酶的抑制影响,但在用抗 HMGB-1 阻断抗体预处理后显著降低。总之,我们的研究结果表明,NET 对气道上皮细胞发挥直接的促炎作用,这可能有助于在肺组织损伤后体内进一步招募中性粒细胞和炎症的持续存在。