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芜湖汤通过 AMPK/ULK1 信号通路调节树突状细胞自噬治疗呼吸道合胞病毒(RSV)诱导的小鼠哮喘

Wuhu Decoction Regulates Dendritic Cell Autophagy in the Treatment of Respiratory Syncytial Virus (RSV)-Induced Mouse Asthma by AMPK/ULK1 Signaling Pathway.

机构信息

Hunan University of Chinese Medicine, Changsha, Hunan, China (mainland).

College of Integrated Traditional Chinese and Western Medicine of Hunan University of Chinese Medicine, Changsha, Hunan, China (mainland).

出版信息

Med Sci Monit. 2019 Jul 20;25:5389-5400. doi: 10.12659/MSM.917692.

Abstract

BACKGROUND Dendritic cell autophagy plays a pivotal role in asthma. Wuhu decoction can significantly improve respiratory syncytial virus (RSV) bronchiolitis and delay its development into asthma. The aim of the present study was to explore the therapeutic effect and mechanism of Wuhu decoction on RSV -induced asthma in mice. MATERIAL AND METHODS Establishment of asthmatic mice model was induced by RSV. Hematoxylin-eosin staining, periodic acid-Schiff (PAS) staining, and Masson trichrome staining were performed to observe pathological changes in the lungs. The levels of CD4⁺ T, CD8⁺ T, and CD4⁺ CD25⁺ T in blood were analyzed by flow cytometry. The contents of interleukin (IL)-4, interferon-gamma (IFN-γ), IL-10, and IL-13 in serum were measured by enzyme-linked immunosorbent assay (ELISA). The number of autophagosomes in dendritic cells (DCs) of lung tissue was observed by transmission electron microscope. The DCs of lung tissue were isolated by magnetic bead sorting. The levels of LC3-II, Beclin-1, and LC3-I in DCs and MMP-9, TIMP-1, AMPK, p-AMPK, ULK1, and LK1 expression in lung tissues were detected by western blot. Real-time polymerase chain reaction (PCR) detected the expression of AMPK and ULK1 genes. RESULTS Wuhu decoction can effectively alleviate chronic airway inflammation and airway remodeling and reduce airway hyperresponsiveness. Moreover, Wuhu decoction can significantly enhance the level of autophagy in DCs of lung tissue and promote the expression of AMPK and ULK1 in lung tissue. CONCLUSIONS Wuhu decoction may improve the RSV-induced asthmatic symptoms by enhancing autophagy of DCs in lung tissue dependent on the AMPK/ULK1 signaling pathway.

摘要

背景

树突状细胞自噬在哮喘中发挥关键作用。芜湖汤可显著改善呼吸道合胞病毒(RSV)细支气管炎,并延缓其发展为哮喘。本研究旨在探讨芜湖汤对 RSV 诱导的哮喘小鼠的治疗作用及机制。方法:采用 RSV 诱导建立哮喘小鼠模型。通过苏木精-伊红(H&E)染色、过碘酸雪夫(PAS)染色和 Masson 三色染色观察肺部病理变化。采用流式细胞术分析血液中 CD4+T、CD8+T 和 CD4+CD25+T 的水平。采用酶联免疫吸附试验(ELISA)检测血清中白细胞介素(IL)-4、干扰素-γ(IFN-γ)、IL-10 和 IL-13 的含量。透射电镜观察肺组织树突状细胞(DC)中的自噬体数量。采用磁珠分选法分离肺组织 DC。采用 Western blot 检测 DC 中 LC3-II、Beclin-1 和 LC3-I 的水平,以及肺组织中 MMP-9、TIMP-1、AMPK、p-AMPK、ULK1 和 LK1 的表达。实时聚合酶链反应(PCR)检测 AMPK 和 ULK1 基因的表达。结果:芜湖汤能有效缓解慢性气道炎症和气道重塑,降低气道高反应性。此外,芜湖汤能显著增强肺组织 DC 中的自噬水平,并促进肺组织中 AMPK 和 ULK1 的表达。结论:芜湖汤可能通过增强肺组织 DC 的自噬作用,改善 RSV 诱导的哮喘症状,其作用机制与 AMPK/ULK1 信号通路有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/10f9/6662944/31e9293e420b/medscimonit-25-5389-g001.jpg

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