hnRNPA2B1促进内皮祖细胞外泌体miR-103-3p的产生，缓解急性呼吸窘迫综合征中巨噬细胞M1极化

IF 4.7 2区医学 Q2 IMMUNOLOGY

International immunopharmacology Pub Date : 2025-05-17 DOI:10.1016/j.intimp.2025.114830

Lei Yang , Ting Chen , Yuanlu Huang , Yuxuan Yang , Xiaoe Cheng , Fusheng Wei

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引用次数: 0

摘要

巨噬细胞极化在急性呼吸窘迫综合征（ARDS）中起着至关重要的作用。近年来，越来越多的证据表明内皮祖细胞（endothelial progenitor cells, EPCs）分泌外泌体（EPCs- exos）在ARDS病理性炎症过程中具有明显的治疗作用，但其潜在机制鲜有报道。方法分离鉴定小鼠原代EPCs和EPCs- exos。PKH-26染色检测RAW264.7细胞对EPCs-Exos的吸收。采用流式细胞术和RT-qPCR分析RAW264.7细胞的极化情况。通过双荧光素酶测定、RNA拉下和RNA免疫共沉淀法验证分子相互作用。建立ARDS小鼠模型，采用HE染色、RT-qPCR和western blotting检测其病理变化及相关分子的表达。结果sepcs - exos能够转移到巨噬细胞中，有效逆转lps诱导的巨噬细胞由M2表型向M1表型的极化；然而，这些变化通过激活TLR4/NF-κB通路而减弱。MiR-103-3p被证明在EPC-Exos中富集，可以通过直接结合TLR4 3′-UTR转移到巨噬细胞并灭活TLR4/NF-κB通路。此外，在lps处理的细胞中，miR-103-3p过表达通过抑制TLR4/NF-κB通路而升高巨噬细胞M2极化，抑制M1极化，而在体外敲低EPC-Exos中miR-103-3p则取消了EPC-Exos对巨噬细胞极化和体内肺炎症损伤的调节作用。HnRNPA2B1被证明与miR-103-3p相互作用，并负责其外泌体分泌，从而抑制促炎巨噬细胞极化。结论hnrnpa2b1介导的EPCs外泌体递送miR-103-3p通过TLR4/NF-κB通路失活，保护ARDS巨噬细胞炎症。

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hnRNPA2B1 promotes the production of exosomal miR-103-3p from endothelial progenitor cells to alleviate macrophage M1 polarization in acute respiratory distress syndrome

Background

Macrophage polarization plays a crucial role in acute respiratory distress syndrome (ARDS). Recently, mounting evidence has uncovered that endothelial progenitor cells (EPCs) secreted exosomes (EPCs-Exos) exert obvious therapeutic effects on the pathological inflammatory process of ARDS, but its potential mechanism is rarely reported.

Methods

The primary mouse EPCs and EPCs-Exos were isolated and identified. Absorption of EPCs-Exos by RAW264.7 cells was examined by PKH-26 staining. The polarization of RAW264.7 cells was evaluated by flow cytometry and RT-qPCR analysis. Molecular interactions were verified by dual luciferase assay, RNA pull-down and RNA immunocoprecipitation assays. ARDS mouse model was established, and pathological changes and expressions of related molecules were detected by HE staining, RT-qPCR and western blotting.

Results

EPCs-Exos could be transferred to macrophages, and effectively reversed LPS-induced polarization of macrophages from M2 to M1 phenotype; however, these changes were diminished by activation of TLR4/NF-κB pathway. MiR-103-3p was proved to be enriched in EPC-Exos and could transfer to macrophage and inactivating TLR4/NF-κB pathway via directly binding to TLR4 3’-UTR. Moreover, miR-103-3p overexpression elevated macrophage M2 polarization and repressed M1 polarization in LPS-treated cells by inhibiting TLR4/NF-κB pathway, and knockdown of miR-103-3p in EPC-Exos abolished the regulatory roles of EPC-Exos on macrophage polarization in vitro, and lung inflammatory injury in vivo. HnRNPA2B1 was proved to interact with miR-103-3p and responsible for its exosomal secretion, which repressed pro-inflammatory macrophage polarization.

Conclusion

These findings suggested that hnRNPA2B1-mediated exosomal delivery of miR-103-3p from EPCs protected against macrophage inflammation in ARDS by inactivation of TLR4/NF-κB pathway.

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来源期刊

International immunopharmacology 医学-免疫学

CiteScore

8.40

自引率

3.60%

发文量

935

审稿时长

53 days

期刊介绍： International Immunopharmacology is the primary vehicle for the publication of original research papers pertinent to the overlapping areas of immunology, pharmacology, cytokine biology, immunotherapy, immunopathology and immunotoxicology. Review articles that encompass these subjects are also welcome. The subject material appropriate for submission includes: • Clinical studies employing immunotherapy of any type including the use of: bacterial and chemical agents; thymic hormones, interferon, lymphokines, etc., in transplantation and diseases such as cancer, immunodeficiency, chronic infection and allergic, inflammatory or autoimmune disorders. • Studies on the mechanisms of action of these agents for specific parameters of immune competence as well as the overall clinical state. • Pre-clinical animal studies and in vitro studies on mechanisms of action with immunopotentiators, immunomodulators, immunoadjuvants and other pharmacological agents active on cells participating in immune or allergic responses. • Pharmacological compounds, microbial products and toxicological agents that affect the lymphoid system, and their mechanisms of action. • Agents that activate genes or modify transcription and translation within the immune response. • Substances activated, generated, or released through immunologic or related pathways that are pharmacologically active. • Production, function and regulation of cytokines and their receptors. • Classical pharmacological studies on the effects of chemokines and bioactive factors released during immunological reactions.