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基于网络药理学和体内实验探讨芜湖汤干预呼吸道合胞病毒诱导哮喘的活性成分及作用机制

[Active components and mechanism of Wuhu Decoction in intervening RSV-induced asthma based on network pharmacology and in vivo experiment].

作者信息

Yao Bing, Liu Jing-Lei, Wang Meng-Qing, Luo Yin-He, Xie Jing, Xun Chun-Zheng, Li Fan, Huang Ting, Zeng Qing-Jia

机构信息

Hunan University of Traditional Chinese Medicine Changsha 410208, China the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine Changsha 410021, China.

Hunan University of Traditional Chinese Medicine Changsha 410208, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2024 Aug;49(15):4148-4157. doi: 10.19540/j.cnki.cjcmm.20240411.406.

Abstract

This study aims to explore the active components and mechanism of Wuhu Decoction in treating respiratory syncytial virus(RSV)-induced asthma. Ultra-high performance liquid chromatography coupled with high-resolution mass spectrometry was used to determine the components of Wuhu Decoction in the blood. By utilizing databases, Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis and Gene Ontology(GO) functional analysis were conducted to investigate the targets of the components of Wuhu Decoction in asthma. Furthermore, the information on target proteins, and metabolite-target-pathway was imported into the STRING database to construct a network interaction diagram to identify the core components and key pathways of Wuhu Decoction. In the in vivo experiment, an asthma model was established using RSV combined with ovalbumin(OVA) in mice. The intervention effect of Wuhu Decoction on RSV-induced asthma in mice was validated through lung function tests, hematoxylin-eosin(HE) staining, enzyme-linked immunosorbent assay(ELISA), Western blot, and immunohistochemistry. The results showed that the main components of Wuhu Decoction in the blood were flavonoids, phenylpropanoids, lignans, and terpenoids. The core components of Wuhu Decoction in treating pediatric asthma included(-)-epigallocatechin, kaempferol, isoliquiritigenin, diosmetin, betulinic acid, ursolic acid, daphnetin, aescin. The main pathways targeted by Wuhu Decoction were calcium signaling pathway, neuroactive ligand-receptor interaction, NOD-like receptor signaling pathway, T cell receptor signaling pathway, and Toll-like receptor signaling pathway. The results of in vivo experiments demonstrated that Wuhu Decoction could improve lung function indicators, down-regulate levels of interleukin-6(IL-6), interleukin-17(IL-17), and tumor necrosis factor-alpha(TNF-α), and reduce the expression of proteins such as NOD-like receptor pyrin domain-containing 3(NLRP3), mitogen-activated protein kinase 1(MAPK1), mitogen-activated protein kinase 14(MAPK14), and nuclear factor kappaB subunit 1(NFKB1) in lung tissue, thereby alleviating neutrophilic inflammation and pulmonary congestion. These findings indicate that Wuhu Decoction intervenes in virus-induced asthma through a synergistic effect on multiple components, targets, and pathways, and it can inhibit the activation of the NOD-like receptor signaling pathway, thereby alleviating airway inflammation and injury in asthmatic mice.

摘要

本研究旨在探讨五虎汤治疗呼吸道合胞病毒(RSV)诱导的哮喘的活性成分及作用机制。采用超高效液相色谱联用高分辨率质谱法测定五虎汤入血成分。利用数据库进行京都基因与基因组百科全书(KEGG)富集分析和基因本体论(GO)功能分析,以研究五虎汤成分在哮喘中的作用靶点。此外,将靶蛋白信息及代谢物-靶标-通路信息导入STRING数据库构建网络相互作用图,以确定五虎汤的核心成分和关键通路。在体内实验中,采用RSV联合卵清蛋白(OVA)建立小鼠哮喘模型。通过肺功能检测、苏木精-伊红(HE)染色、酶联免疫吸附测定(ELISA)、蛋白质印迹法和免疫组织化学法验证五虎汤对RSV诱导的小鼠哮喘的干预效果。结果表明,五虎汤入血的主要成分有黄酮类、苯丙素类、木脂素类和萜类。五虎汤治疗小儿哮喘的核心成分包括(-)-表没食子儿茶素、山柰酚、异甘草素、香叶木素、桦木酸、熊果酸、瑞香素、七叶皂苷。五虎汤主要作用的通路有钙信号通路、神经活性配体-受体相互作用、NOD样受体信号通路、T细胞受体信号通路和Toll样受体信号通路。体内实验结果表明,五虎汤可改善肺功能指标,下调白细胞介素-6(IL-6)、白细胞介素-17(IL-17)和肿瘤坏死因子-α(TNF-α)水平,并降低肺组织中含NOD样受体吡喃结构域蛋白3(NLRP3)、丝裂原活化蛋白激酶1(MAPK1)、丝裂原活化蛋白激酶14(MAPK14)和核因子κB亚基1(NFKB1)等蛋白的表达,从而减轻中性粒细胞炎症和肺淤血。这些研究结果表明,五虎汤通过对多种成分、靶点和通路的协同作用干预病毒诱导的哮喘,且能抑制NOD样受体信号通路的激活,从而减轻哮喘小鼠的气道炎症和损伤。

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