吡非尼酮通过抑制中性粒细胞胞外陷阱形成和NLRP3炎性小体激活来减轻小鼠间质性肺疾病。

Pirfenidone alleviates interstitial lung disease in mice by inhibiting neutrophil extracellular trap formation and NLRP3 inflammasome activation.

作者信息

Su Qiyan, Feng Yingyue, Guo Jin, Cui Xi, Zhu Jiarui, Yang Jumei, Zhang Sigong

机构信息

The Second Clinical Medical College, Lanzhou University, Lanzhou, Gansu, China.

Department of Rheumatology, Lanzhou University Second Hospital, Lanzhou, Gansu, China.

出版信息

Clin Exp Immunol. 2025 Jan 21;219(1). doi: 10.1093/cei/uxaf019.

DOI:10.1093/cei/uxaf019

PMID:40117382

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12038160/

Abstract

BACKGROUND

Idiopathic inflammatory myopathy (IIM) is a progressive autoimmune disease characterized by interstitial lung disease (ILD) with limited therapeutics available. Pirfenidone (PFD), a medication utilized for the treatment of idiopathic pulmonary fibrosis, exhibits notable antioxidant, anti-inflammatory, and inhibition of collagen synthesis. This study aims to clarify its efficacy and mechanism in treating IIM-ILD.

METHODS

A murine myositis-associated interstitial lung disease (MAILD) model was used to assess the therapeutic effect of PFD. The serum levels of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). Pirfenidone was utilized to disrupt neutrophil extracellular traps (NETs) formation in vitro, and its inhibitory effect on NETs was assessed through immunohistochemistry of citrullinated histone H3 and myeloperoxidase in the lung tissue and the serum cfDNA level in mice. Immunohistochemical and western blot were utilized to examine alterations in epithelial-mesenchymal transition (EMT) and NOD-like receptor protein 3 (NLRP3) inflammasome markers.

RESULTS

Pirfenidone treatment inhibited pulmonary inflammation and fibrosis in the MAILD model. Pirfenidone intervention reduced NETs formation in vitro. Pirfenidone treatment significantly reduces NETs infiltration in the lung tissue and the level of cfDNA in the serum of mice. Additionally, PFD downregulated EMT and NLRP3-related proteins in vivo. Pirfenidone treatment also notably reduced serum levels of IL-1β, IL-6, and TNF-α. After NETs stimulation, A549 cells exhibited EMT and activation of NLRP3 inflammasome. Pirfenidone attenuated EMT in A549 cells and suppressed the activation of NLRP3 inflammasome.

CONCLUSION

Pirfenidone alleviates ILD in a murine MAILD model by inhibiting NETs formation and NLRP3 inflammasome activation, suggesting that PFD might be a potential therapeutic agent for IIM-ILD.

摘要

背景

特发性炎症性肌病（IIM）是一种进行性自身免疫性疾病，其特征为间质性肺疾病（ILD），可用的治疗方法有限。吡非尼酮（PFD）是一种用于治疗特发性肺纤维化的药物，具有显著的抗氧化、抗炎和抑制胶原蛋白合成的作用。本研究旨在阐明其治疗IIM-ILD的疗效和机制。

方法

使用小鼠肌炎相关性间质性肺疾病（MAILD）模型评估PFD的治疗效果。通过酶联免疫吸附测定（ELISA）检测白细胞介素（IL）-1β、IL-6和肿瘤坏死因子-α（TNF-α）的血清水平。在体外使用吡非尼酮破坏中性粒细胞胞外陷阱（NETs）的形成，并通过对肺组织中瓜氨酸化组蛋白H3和髓过氧化物酶进行免疫组织化学以及检测小鼠血清cfDNA水平来评估其对NETs的抑制作用。利用免疫组织化学和蛋白质免疫印迹法检测上皮-间质转化（EMT）和NOD样受体蛋白3（NLRP3）炎性小体标志物的变化。

结果

吡非尼酮治疗可抑制MAILD模型中的肺部炎症和纤维化。吡非尼酮干预可减少体外NETs的形成。吡非尼酮治疗可显著减少小鼠肺组织中NETs的浸润以及血清中cfDNA的水平。此外，PFD在体内下调了EMT和NLRP3相关蛋白。吡非尼酮治疗还显著降低了血清中IL-1β、IL-6和TNF-α的水平。NETs刺激后，A549细胞表现出EMT和NLRP3炎性小体的激活。吡非尼酮减弱了A549细胞中的EMT并抑制了NLRP3炎性小体的激活。