Neutrophil extracellular traps induce pyroptosis of pulmonary microvascular endothelial cells by activating the NLRP3 inflammasome.

IF 3.4 3区 医学 Q3 IMMUNOLOGY
Peipei Zhao, Jiarui Zhu, Ling Bai, Wenlan Ma, Feifei Li, Cen Zhang, Liangtao Zhao, Liuyang Wang, Sigong Zhang
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Abstract

Excessive formation of neutrophil extracellular traps (NETs) may lead to myositis-related interstitial lung disease (ILD). There is evidence that NETs can directly injure vascular endothelial cells and play a pathogenic role in the inflammatory exudation of ILD. However, the specific mechanism is unclear. This study aimed to investigate the specific mechanism underlying NET-induced injury to human pulmonary microvascular endothelial cells (HPMECs). HPMECs were stimulated with NETs (200 ng/ml) in vitro. Cell death was detected by propidium iodide staining. The morphological changes of the cells were observed by transmission electron microscopy (TEM). Pyroptosis markers were detected by western blot, immunofluorescence, and quantitative real-time polymerase chain reaction, and the related inflammatory factor Interleukin-1β (IL-1β) was verified by enzyme-linked immunosorbent assay (ELISA). Compared with the control group, HPMECs mortality increased after NET stimulation, and the number of pyroptosis vacuoles in HPMECs was further observed by TEM. The pulmonary microvascular endothelial cells (PMECs) of the experimental autoimmune myositis mouse model also showed a trend of pyroptosis in vivo. Cell experiment further confirmed the significantly high expression of the NLRP3 inflammasome and pyroptosis-related markers, including GSDMD and inflammatory factor IL-1β. Pretreated with the NLRP3 inhibitor MCC950, the activation of NLRP3 inflammasome and pyroptosis of HPMECs were effectively inhibited. Our study confirmed that NETs promote pulmonary microvascular endothelial pyroptosis by activating the NLRP3 inflammasome, suggesting that NETs-induced pyroptosis of PMECs may be a potential pathogenic mechanism of inflammatory exudation in ILD.

中性粒细胞胞外捕获物通过激活 NLRP3 炎症小体诱导肺微血管内皮细胞发生脓毒症。
中性粒细胞胞外捕获物(NET)的过度形成可能会导致肌炎相关的间质性肺病(ILD)。有证据表明,NET 可直接损伤血管内皮细胞,并在 ILD 的炎性渗出中发挥致病作用。然而,其具体机制尚不清楚。本研究旨在探讨NET诱导的人肺微血管内皮细胞(HPMECs)损伤的具体机制。在体外用 NET(200 ng/ml)刺激 HPMECs。通过碘化丙啶染色检测细胞死亡。透射电子显微镜(TEM)观察细胞的形态变化。通过Western印迹、免疫荧光和实时定量PCR检测了炭疽标志物,并通过ELISA检测了相关炎症因子IL-1β。与对照组相比,NET刺激后HPMECs死亡率升高,TEM进一步观察到HPMECs中热凋亡空泡的数量。实验性自身免疫性肌炎(EAM)小鼠模型的肺微血管内皮细胞(PMECs)在体内也出现了化脓趋势。细胞实验进一步证实了 NLRP3 炎性体和热噬相关标记物(包括 GSDMD 和炎性因子 IL-1β)的明显高表达。用NLRP3抑制剂MCC950预处理后,HPMECs的NLRP3炎性体活化和化脓过程被有效抑制。我们的研究证实,NET通过激活NLRP3炎性体促进肺微血管内皮细胞的热凋亡,提示NET诱导的PMECs热凋亡可能是ILD炎性渗出的潜在致病机制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
8.40
自引率
2.20%
发文量
101
审稿时长
3-8 weeks
期刊介绍: Clinical & Experimental Immunology (established in 1966) is an authoritative international journal publishing high-quality research studies in translational and clinical immunology that have the potential to transform our understanding of the immunopathology of human disease and/or change clinical practice. The journal is focused on translational and clinical immunology and is among the foremost journals in this field, attracting high-quality papers from across the world. Translation is viewed as a process of applying ideas, insights and discoveries generated through scientific studies to the treatment, prevention or diagnosis of human disease. Clinical immunology has evolved as a field to encompass the application of state-of-the-art technologies such as next-generation sequencing, metagenomics and high-dimensional phenotyping to understand mechanisms that govern the outcomes of clinical trials.
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