肌炎相关ILD中TLR9信号参与NETs诱导的肺泡上皮间质转化  

Involvement of Toll-like receptor 9 signaling in neutrophil extracellular traps-induced epithelial-to-mesenchymal transition in myositis-associated interstitial lung disease

作　　者：冯映月 禚萃 苏其燕 崔曦 郭瑾 朱嘉睿 张思功[3] Feng Yingyue;Zhuo Cui;Su Qiyan;Cui Xi;Guo Jin;Zhu Jiarui;Zhang Sigong(The Second Hospital&Clinical Medical School,Lanzhou University,Lanzhou 730030,Gansu,China;Cuiying Biomedical Research Center,Lanzhou University Second Hospital,Lanzhou 730030,Gansu,China;Department of Rheumatology and Immunology,the Second Hospital&Clinical Medical School,Lanzhou University,Lanzhou 730030,Gansu,China)

机构地区：[1]兰州大学第二医院(第二临床医学院),甘肃兰州730030 [2]兰州大学第二医院萃英生物医学研究中心,甘肃兰州730030 [3]兰州大学第二医院(第二临床医学院)风湿免疫科,甘肃兰州730030

出　　处：《生物医学转化》2024年第4期87-95,共9页Biomedical Transformation

基　　金：国家自然科学基金地区基金项目(82060302,82260325);甘肃省自然科学基金联合基金项目(24JRRA923);兰州大学医学创新与发展优秀青年人才项目(lzuyxcx-2022-168);兰州大学“中央高校基本科研业务费专项资金”杰出青年支持计划项目(lzujbky-2024-oy03);甘肃省中医药科研项目(GZKZ-2024-29)。

摘　　要：目的 旨在阐明中性粒细胞胞外诱捕网(NETs)是否通过Toll样受体9 (TLR9)信号促进肺泡上皮细胞间质转化(EMT)促进特发性肌炎相关间质性肺病(IIM-ILD)的发生。方法 采用实验性自身免疫性肌炎(EAM)小鼠模型,使用组织病理学、血清学方法明确IIM-ILD中是否存在NETs浸润、EMT表型及TLR9激活;采用NETs体外干预A549细胞,转录组测序、免疫荧光、蛋白质印迹法等方法,探究NETs诱导EMT的机制。结果 EAM小鼠肺组织存在NETs浸润、EMT现象及TLR9信号激活。体外实验中NETs影响A549细胞活力及形态,激活TLR9信号通路,诱导EMT的发生,并诱导促炎和促纤维化相关基因上调,其中TLR9基因显著高表达,而TLR9抑制剂减轻了EMT。结论 NETs诱导的EMT可能是IIM-ILD的重要致病机制,其中TLR9信号起关键作用,靶向TLR9信号可能是IIM-ILD的治疗方向。Objective The study aims to elucidate whether neutrophil extracellular traps(NETs) promote epithelial-mesenchymal transition(EMT) in alveolar epithelial cells via Toll-like receptor 9(TLR9) signaling,thereby contributing to the development of idiopathic inflammatory myopathy-associated interstitial lung disease(IIM-ILD).Methods An experimental autoimmune myositis(EAM) mouse model was used,using histological and serological methods to clearly define the presence of NETs infiltration,EMT phenotypes,and TLR9 activation in IIM-ILD.In vitro,A549 cells were treated with NETs,and methods such as transcriptome sequencing,immunofluorescence,and Western blotting were used to investigate the mechanisms underlying NETs-induced EMT.Results Lung tissues from EAM mice exhibited NETs infiltration,EMT,and activation of TLR9 signaling.In vitro experiments demonstrated that NETs affected A549 cell viability and morphology,activated the TLR9signaling pathway,and induced EMT.Moreover,NETs upregulated genes associated with inflammation and fibrosis.Notably,TLR9 gene expression was significantly elevated,and the use of a TLR9 inhibitor alleviated EMT.Conclusion NETs-induced EMT may serve as a key pathogenic mechanism in IIM-ILD,with TLR9signaling playing a key role.Targeting TLR9 signaling may offer a promising therapeutic approach for IIM-ILD.

关 键 词：中性粒细胞胞外诱捕网 特发性肌炎 间质性肺病 上皮间质转化 TOLL样受体9 

分 类 号：R593.2[医药卫生—内科学]