Hydrogen sulfide (H₂S) is a toxic air pollutant that causes immune damage. Recent studies have found that neutrophil extracellular trap (NET) formation is one way in which neutrophils exert immune functions. In addition, the formation of NETs is also related to thrombosis and autoimmune diseases. Recent studies have shown that miRNAs are involved in the regulation of a variety of pathophysiological processes. Here, we investigated the role of H₂S in regulating the formation of NETs by affecting miR-16–5p. Our study established an in vitro H₂S exposure model for neutrophils using phorbol-myristate-acetate (PMA) to induce NET formation. We observed the morphological changes of cells with scanning electron microscopy and fluorescence microscopy. Then, the content of extracellular DNA and the expression of MPO and NE in each group were detected. The results showed that H₂S inhibited the formation of NETs. The expression of miR-16–5p and its target genes PiK3R1 and RAF1 was then measured by qRT-PCR. H₂S upregulated miR-16–5p and inhibited expression of the target genes PiK3R1 and RAF1, and it subsequently inhibited the Pi3K/AKT and ERK pathways and decreased respiratory burst levels. Furthermore, H₂S attenuated inositol 1,4,5-trisphosphate receptor (IP3R)-mediated endoplasmic reticulum calcium outflow as well as autophagy caused by PMA. This study enriches H₂S immunotoxicity research and provides a possible solution for the treatment of NET-related diseases.

硫化氢（H ₂ S）是一种有毒的空气污染物，会引起免疫损伤。最近的研究发现，嗜中性白细胞胞外陷阱（NET）的形成是嗜中性白细胞发挥免疫功能的一种方式。此外，NETs的形成还与血栓形成和自身免疫性疾病有关。最近的研究表明，miRNA参与了多种病理生理过程的调控。在这里，我们研究了H ₂ S通过影响miR-16-5p在调节NETs形成中的作用。我们的研究建立了体外H ₂中性粒细胞的S暴露模型，使用佛波-肉豆蔻酸酯-乙酸酯（PMA）诱导NET的形成。我们用扫描电子显微镜和荧光显微镜观察了细胞的形态变化。然后，检测每组中细胞外DNA的含量以及MPO和NE的表达。结果表明，H ₂ S抑制了NETs的形成。然后通过qRT-PCR测量miR-16-5p及其靶基因PiK3R1和RAF1的表达。H ₂ S上调了miR-16-5p并抑制了靶基因PiK3R1和RAF1的表达，随后它抑制了Pi3K / AKT和ERK途径并降低了呼吸爆发水平。此外，H ₂S减弱了肌醇1,4,5-三磷酸受体（IP3R）介导的内质网钙流出以及PMA引起的自噬。这项研究丰富了H ₂ S免疫毒性研究，并为治疗NET相关疾病提供了可能的解决方案。