Institute of Animal Sciences of Chinese Academy of Agricultural Sciences, Number 2, Yuanmingyuan West Road, Haidian District, Beijing, 100193, PR China; College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, PR China.
College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, PR China.
Ecotoxicol Environ Saf. 2020 May;194:110412. doi: 10.1016/j.ecoenv.2020.110412. Epub 2020 Mar 7.
Hydrogen sulfide (HS) 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 HS in regulating the formation of NETs by affecting miR-16-5p. Our study established an in vitro HS 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 HS inhibited the formation of NETs. The expression of miR-16-5p and its target genes PiK3R1 and RAF1 was then measured by qRT-PCR. HS 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, HS attenuated inositol 1,4,5-trisphosphate receptor (IP3R)-mediated endoplasmic reticulum calcium outflow as well as autophagy caused by PMA. This study enriches HS immunotoxicity research and provides a possible solution for the treatment of NET-related diseases.
硫化氢（HS）是一种有毒的空气污染物，可导致免疫损伤。最近的研究发现，中性粒细胞胞外诱捕网（NET）的形成是中性粒细胞发挥免疫功能的一种方式。此外，NET 的形成也与血栓形成和自身免疫性疾病有关。最近的研究表明，miRNAs 参与调节多种病理生理过程。在这里，我们研究了 HS 通过影响 miR-16-5p 调节 NET 形成的作用。我们使用佛波醇肉豆蔻酸乙酸酯（PMA）建立了体外 HS 暴露中性粒细胞模型，以诱导 NET 形成。我们通过扫描电子显微镜和荧光显微镜观察细胞的形态变化。然后，检测各组细胞外 DNA 的含量以及 MPO 和 NE 的表达。结果表明，HS 抑制 NET 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。HS 上调了 miR-16-5p，抑制了靶基因 PiK3R1 和 RAF1 的表达，继而抑制了 Pi3K/AKT 和 ERK 通路，并降低了呼吸爆发水平。此外，HS 减弱了 PMA 诱导的肌醇 1,4,5-三磷酸受体（IP3R）介导的内质网钙外流和自噬。本研究丰富了 HS 免疫毒性研究，并为治疗 NET 相关疾病提供了一种可能的解决方案。
