淫羊藿苷Ⅱ通过调节慢性哮喘中的炎症小体和促纤维化信号靶向M2巨噬细胞
2026/02/02
摘要
背景：慢性哮喘的特征是持续性气道炎症、气道高反应性和结构重塑。巨噬细胞，尤其是交替激活的M2细胞，是2型免疫和纤维化的关键驱动因素。淫羊藿苷Ⅱ（ICAII）是一种从淫羊中藿提取的黄酮类化合物，具有已知的抗炎特性，但其对哮喘中M2巨噬细胞的确切免疫调节作用仍不清楚。
目的：本研究旨在利用模拟慢性哮喘的小鼠模型评估ICAII的潜在治疗作用，并明确其潜在机制，重点关注巨噬细胞极化和免疫介导的组织重塑。
方法：通过卵清蛋白（OVA）致敏和反复暴露在BALB/c小鼠中建立慢性哮喘模型，随后给予不同剂量的ICAII。采用肺功能测试、组织学分析、酶联免疫吸附测定（ELISA）、流式细胞术和免疫组织化学方法评估炎症、气道重塑和巨噬细胞极化情况。使用RAW264.7和MHS巨噬细胞系进行的体外实验进一步研究了ICAII对M2分化的影响。整合转录组测序、网络药理学、分子对接分析和验证以确定关键调控通路。
结果：ICAII改善了气道阻力和顺应性，减轻了炎症浸润和胶原沉积，降低了Th2细胞因子、血清IgE和促纤维化标志物水平，在40mg/kg剂量下效果最为显著。在体内，ICAII抑制了M2巨噬细胞的积累；在体外，它抑制了M2分化，同时对M1标志物表达有不同的影响。网络药理学和分子对接预测ICAII与SIRT1之间存在适度亲和力的相互作用，表面等离子体共振（SPR）和酶活性测定实验证实了这一点。结合转录组学和药理学分析，这些结果确定了SIRT1/NLRP3和TGF - β/Smad3/VEGF轴是介导ICAII对慢性哮喘保护作用的主要通路。具体而言，SIRT1/NLRP3轴指的是沉默调节蛋白1（SIRT1）介导的对含NLR家族吡啶结构域3（NLRP3）炎症小体激活的抑制，而TGF - β/Smad3/VEGF轴代表与血管内皮生长因子相关的转化生长因子β驱动的促纤维化信号级联反应。从机制上讲，ICAII增强了SIRT1的活性，抑制了与NLRP3炎症小体相关的炎症，抑制了促纤维化的TGF - β/Smad3信号级联反应及其与重塑相关的下游效应因子VEGF，从而抑制了M2巨噬细胞极化和组织重塑。
结论：我们首次在慢性哮喘中提供了综合机制证据，表明ICAII通过双轴策略重新编程M2巨噬细胞，同时激活SIRT1以抑制NLRP3炎症小体并减轻TGF - β/Smad3/VEGF驱动的重塑。与以往仅描述ICAII广泛抗炎作用的报道不同，我们的研究独特地将免疫调节与结构保护联系起来，并进一步提供了直接的靶点结合验证。这种药物－靶点－功能的连续性不仅描绘了以巨噬细胞为中心的慢性哮喘病理生物学范式，而且引入了一种具有转化应用价值的机制，以解决长期存在的持续性气道重塑和T2高炎症的挑战。
Icariside II targets M2 macrophages by regulating the inflammasome and profibrotic signaling in chronic asthma
Mengmeng Chen, Huijie Zhang, Congcong Li, Jiaqi Liu, Huahe Zhu, Weifeng Tang, Xueyi Zhu, Jiemin He, Yuting Shi, Yalikun Maimaititusun, Na Wang, Jingcheng Dong
Abstract
Background: Chronic asthma features persistent airway inflammation, airway hyper-responsiveness, and structural remodeling. Macrophages, especially alternatively activated M2 cells, are key drivers of type-2 immunity and fibrosis. Icariside II (ICAII), a flavonoid from Epimedium, exhibits known anti-inflammatory properties, but its precise immunomodulatory effects on M2 macrophages in asthma remain elusive.
Objective: This work sought to evaluate the potential therapeutic role of ICAII using a mouse model that mimics chronic asthma and define the underlying mechanisms, with emphasis on macrophage polarization and immune-mediated tissue remodeling.
Methods: Chronic asthma was established in BALB/c mice through ovalbumin (OVA) sensitization and repeated exposure, followed by ICAII administration at various doses. Pulmonary function tests, histological analyses, ELISA, flow cytometry, and immunohistochemistry were employed to assess inflammation, airway remodeling, and macrophage polarization. In vitro experiments using RAW264.7 and MHS macrophage lines further investigated ICAII's impact on M2 differentiation. Transcriptome sequencing, network pharmacology, molecular docking analyses, and validation were integrated to identify key regulatory pathways.
Results: ICAII improved airway resistance and compliance, alleviated inflammatory infiltration and collagen deposition, and lowered Th2 cytokines, serum IgE, and pro-fibrotic markers, with the most pronounced effects observed at 40 mg/kg. In vivo, ICAII suppressed M2 macrophage accumulation, and in vitro, it inhibited M2 differentiation, while with a divergent impact on M1 marker expression. Network pharmacology and molecular docking predicted a moderate affinity interaction between ICAII and SIRT1, which was experimentally confirmed by SPR and enzymatic activity assays. Combined with transcriptomic and pharmacological analyses, these results identified the SIRT1/NLRP3 and TGF-β/Smad3/VEGF axes as principal pathways mediating the protective effects of ICAII against chronic asthma. Specifically, the SIRT1/NLRP3 axis refers to Sirtuin1 (SIRT1)-mediated suppression of NLR family pyrin domain containing 3 (NLRP3) inflammasome activation, whereas the TGF-β/Smad3/VEGF axis represents a transforming growth factor beta-driven profibrotic signaling cascade associated with vascular endothelial growth factor. Mechanistically, ICAII enhanced SIRT1 activity, suppressed NLRP3 inflammasome-associated inflammation, and inhibited the profibrotic TGF-β/Smad3 signaling cascade and its remodeling-associated downstream effector VEGF, thereby restraining M2 macrophage polarization and tissue remodeling.
Conclusion: We provide the first integrated mechanistic evidence in chronic asthma that ICAII reprograms M2 macrophages via a dual-axis strategy, simultaneously activating SIRT1 to suppress the NLRP3 inflammasome and attenuating TGF-β/Smad3/VEGF-driven remodeling. Unlike previous reports that only described the broad anti-inflammatory effects of ICAII, our study uniquely links immune modulation with structural protection and further delivers direct target engagement validation. This drug-target-function continuum not only delineates a macrophage-centered paradigm for chronic asthma pathobiology but also introduces a translationally actionable mechanism to address the long-standing challenge of persistent airway remodeling and T2-high inflammation.
背景：慢性哮喘的特征是持续性气道炎症、气道高反应性和结构重塑。巨噬细胞，尤其是交替激活的M2细胞，是2型免疫和纤维化的关键驱动因素。淫羊藿苷Ⅱ（ICAII）是一种从淫羊中藿提取的黄酮类化合物，具有已知的抗炎特性，但其对哮喘中M2巨噬细胞的确切免疫调节作用仍不清楚。
目的：本研究旨在利用模拟慢性哮喘的小鼠模型评估ICAII的潜在治疗作用，并明确其潜在机制，重点关注巨噬细胞极化和免疫介导的组织重塑。
方法：通过卵清蛋白（OVA）致敏和反复暴露在BALB/c小鼠中建立慢性哮喘模型，随后给予不同剂量的ICAII。采用肺功能测试、组织学分析、酶联免疫吸附测定（ELISA）、流式细胞术和免疫组织化学方法评估炎症、气道重塑和巨噬细胞极化情况。使用RAW264.7和MHS巨噬细胞系进行的体外实验进一步研究了ICAII对M2分化的影响。整合转录组测序、网络药理学、分子对接分析和验证以确定关键调控通路。
结果：ICAII改善了气道阻力和顺应性，减轻了炎症浸润和胶原沉积，降低了Th2细胞因子、血清IgE和促纤维化标志物水平，在40mg/kg剂量下效果最为显著。在体内，ICAII抑制了M2巨噬细胞的积累；在体外，它抑制了M2分化，同时对M1标志物表达有不同的影响。网络药理学和分子对接预测ICAII与SIRT1之间存在适度亲和力的相互作用，表面等离子体共振（SPR）和酶活性测定实验证实了这一点。结合转录组学和药理学分析，这些结果确定了SIRT1/NLRP3和TGF - β/Smad3/VEGF轴是介导ICAII对慢性哮喘保护作用的主要通路。具体而言，SIRT1/NLRP3轴指的是沉默调节蛋白1（SIRT1）介导的对含NLR家族吡啶结构域3（NLRP3）炎症小体激活的抑制，而TGF - β/Smad3/VEGF轴代表与血管内皮生长因子相关的转化生长因子β驱动的促纤维化信号级联反应。从机制上讲，ICAII增强了SIRT1的活性，抑制了与NLRP3炎症小体相关的炎症，抑制了促纤维化的TGF - β/Smad3信号级联反应及其与重塑相关的下游效应因子VEGF，从而抑制了M2巨噬细胞极化和组织重塑。
结论：我们首次在慢性哮喘中提供了综合机制证据，表明ICAII通过双轴策略重新编程M2巨噬细胞，同时激活SIRT1以抑制NLRP3炎症小体并减轻TGF - β/Smad3/VEGF驱动的重塑。与以往仅描述ICAII广泛抗炎作用的报道不同，我们的研究独特地将免疫调节与结构保护联系起来，并进一步提供了直接的靶点结合验证。这种药物－靶点－功能的连续性不仅描绘了以巨噬细胞为中心的慢性哮喘病理生物学范式，而且引入了一种具有转化应用价值的机制，以解决长期存在的持续性气道重塑和T2高炎症的挑战。
（北京朝阳医院呼吸与危重症医学科 顾宪民 摘译 中日友好医院呼吸与危重症医学科 林江涛 审校）
（Phytomedicine. 2026 Jan 6:151:157787. doi: 10.1016/j.phymed.2026.157787. Online ahead of print.）
（Phytomedicine. 2026 Jan 6:151:157787. doi: 10.1016/j.phymed.2026.157787. Online ahead of print.）
Icariside II targets M2 macrophages by regulating the inflammasome and profibrotic signaling in chronic asthma
Mengmeng Chen, Huijie Zhang, Congcong Li, Jiaqi Liu, Huahe Zhu, Weifeng Tang, Xueyi Zhu, Jiemin He, Yuting Shi, Yalikun Maimaititusun, Na Wang, Jingcheng Dong
Abstract
Background: Chronic asthma features persistent airway inflammation, airway hyper-responsiveness, and structural remodeling. Macrophages, especially alternatively activated M2 cells, are key drivers of type-2 immunity and fibrosis. Icariside II (ICAII), a flavonoid from Epimedium, exhibits known anti-inflammatory properties, but its precise immunomodulatory effects on M2 macrophages in asthma remain elusive.
Objective: This work sought to evaluate the potential therapeutic role of ICAII using a mouse model that mimics chronic asthma and define the underlying mechanisms, with emphasis on macrophage polarization and immune-mediated tissue remodeling.
Methods: Chronic asthma was established in BALB/c mice through ovalbumin (OVA) sensitization and repeated exposure, followed by ICAII administration at various doses. Pulmonary function tests, histological analyses, ELISA, flow cytometry, and immunohistochemistry were employed to assess inflammation, airway remodeling, and macrophage polarization. In vitro experiments using RAW264.7 and MHS macrophage lines further investigated ICAII's impact on M2 differentiation. Transcriptome sequencing, network pharmacology, molecular docking analyses, and validation were integrated to identify key regulatory pathways.
Results: ICAII improved airway resistance and compliance, alleviated inflammatory infiltration and collagen deposition, and lowered Th2 cytokines, serum IgE, and pro-fibrotic markers, with the most pronounced effects observed at 40 mg/kg. In vivo, ICAII suppressed M2 macrophage accumulation, and in vitro, it inhibited M2 differentiation, while with a divergent impact on M1 marker expression. Network pharmacology and molecular docking predicted a moderate affinity interaction between ICAII and SIRT1, which was experimentally confirmed by SPR and enzymatic activity assays. Combined with transcriptomic and pharmacological analyses, these results identified the SIRT1/NLRP3 and TGF-β/Smad3/VEGF axes as principal pathways mediating the protective effects of ICAII against chronic asthma. Specifically, the SIRT1/NLRP3 axis refers to Sirtuin1 (SIRT1)-mediated suppression of NLR family pyrin domain containing 3 (NLRP3) inflammasome activation, whereas the TGF-β/Smad3/VEGF axis represents a transforming growth factor beta-driven profibrotic signaling cascade associated with vascular endothelial growth factor. Mechanistically, ICAII enhanced SIRT1 activity, suppressed NLRP3 inflammasome-associated inflammation, and inhibited the profibrotic TGF-β/Smad3 signaling cascade and its remodeling-associated downstream effector VEGF, thereby restraining M2 macrophage polarization and tissue remodeling.
Conclusion: We provide the first integrated mechanistic evidence in chronic asthma that ICAII reprograms M2 macrophages via a dual-axis strategy, simultaneously activating SIRT1 to suppress the NLRP3 inflammasome and attenuating TGF-β/Smad3/VEGF-driven remodeling. Unlike previous reports that only described the broad anti-inflammatory effects of ICAII, our study uniquely links immune modulation with structural protection and further delivers direct target engagement validation. This drug-target-function continuum not only delineates a macrophage-centered paradigm for chronic asthma pathobiology but also introduces a translationally actionable mechanism to address the long-standing challenge of persistent airway remodeling and T2-high inflammation.
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在ATLANTIS队列中通过聚类分析识别不同的哮喘表型
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通过神经系统将慢性疼痛遗传学与迟发性哮喘联系起来的因果证据
