Neutrophil extracellular trap (NET) formation eliminates/prevents the spread of infectious agents. Platelet activating factor (PAF) is involved in infectious diseases of cattle because it recruits and activates neutrophils. However, its ability to induce NET release and the role of metabolism in this process is not known. We investigated if inhibition of glycolysis, mitochondrial-derived adenosine triphosphate (ATP) synthesis and purinergic signaling though P2X₁ purinoceptors interfered with NET formation induced by PAF. We inhibited bovine neutrophils with 2-deoxy-d-glucose, rotenone, carbonyl cyanide 3-chlorophenylhydrazone (CCCP) and NF449 to evaluate PAF-mediated NET extrusion. PAF induced mitochondrial hyperpolarization and triggered extracellular ATP release via pannexin-1. Inhibition of mitochondrial metabolism prevented extracellular ATP release. Inhibition of glycolysis, complex-I activity and oxidative phosphorylation prevented NET formation induced by PAF. Inhibition of P2X₁ purinergic receptors inhibited mitochondrial hyperpolarization and NET formation. We concluded that PAF-induced NET release is dependent upon glycolysis, mitochondrial ATP synthesis and purinergic signaling.

中性粒细胞胞外陷阱 (NET) 的形成消除/防止传染源的传播。血小板激活因子 (PAF) 与牛的传染病有关，因为它会招募和激活中性粒细胞。然而，其诱导 NET 释放的能力以及代谢在该过程中的作用尚不清楚。我们研究了通过 P2X ₁嘌呤受体抑制糖酵解、线粒体衍生的三磷酸腺苷 (ATP) 合成和嘌呤能信号传导是否会干扰 PAF 诱导的 NET 形成。我们抑制牛中性粒细胞与2-脱氧d葡萄糖，鱼藤酮，氰化物羰3-氯苯腙（CCCP）和NF449评价PAF介导的NET挤出。PAF诱导的线粒体超极化和触发细胞外ATP释放通过pannexin-1。线粒体代谢的抑制阻止了细胞外 ATP 的释放。糖酵解、复合物 I 活性和氧化磷酸化的抑制阻止了 PAF 诱导的 NET 形成。抑制 P2X ₁嘌呤能受体可抑制线粒体超极化和 NET 形成。我们得出结论，PAF 诱导的 NET 释放取决于糖酵解、线粒体 ATP 合成和嘌呤能信号传导。