Abstract：
BACKGROUND: Jiawei Yanghe Decoction (JWYHD) is modified Yanghe Decoction (YHD). YHD historically utilized as a potent medicinal solution for addressing chronic inflammatory conditions, holds promising therapeutic potential in the treatment of asthma. However, the mechanisms underlying JWYHD\'s effects on allergic asthma remain unclear.
OBJECTIVE: To investigate the therapeutic effect as well as the underlying mechanisms of JWYHD on asthmatic mice.
METHODS: The ovalbumin (OVA)-induced mouse model was utilized, followed by the administration of JWYHD to allergic asthmatic mice. Subsequently, inflammatory cells in the bronchoalveolar lavage fluid (BALF) and lung tissues were conducted. The levels of various cytokines including interleukin (IL)-4, IL-5, IL-13, IL-33, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ in BALF, as well as the total immunoglobulin E (IgE) content in serum, were assessed. Lung function and tissue pathology examinations were performed to assess the protective impacts of JWYHD. The chemical components of JWYHD and its lung prototype compounds (referred to the chemical components present in JWYHD that were observed in the lung) were explored by ultra-high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). RNA-seq analysis revealed the regulation mechanisms of JWYHD treating asthma. Furthermore, the effect of JWYHD on type 2 innate lymphoid cells (ILC2s) in asthmatic mice was detected by flow cytometry and Smart-RNA-seq analysis. Then molecular docking analysis was used to show the interaction between identified compounds and key targets.
RESULTS: JWYHD significantly attenuated the airway inflammation of asthmatic mice, reduced the levels of inflammatory cells in BALF, as well the levels of the cytokines IL-4, IL-5, IL-13, IL-33, and TNF-α in BALF and IgE in serum. Airway hyperresponsiveness (AHR) and lung inflammation infiltration were also alleviated by JWYHD. Moreover, RNA-seq analysis revealed that JWYHD attenuated airway inflammation in asthmatic mice via regulating immunity. Flow cytometry confirmed that JWYHD could inhibit ILC2 responses. ILC2 Smart-RNA-seq analysis showed that JWYHD impaired the inflammation reaction-related signaling pathways in ILC2s, and neuropilin-1 (Nrp1), endothelial transcription factor 3 (GATA3) and interleukin 1 receptor like protein 1 (ST2) might be the key targets. The molecular docking analysis investigating the connection between the primary targets and JWYHD\'s prototype compounds in the lung demonstrated that liquiritin apioside, icariin, glycyrrhizic acid, and uralsaponin B, identified through UPLC-Q-TOF/MS, exhibited significant affinity in binding to the mentioned key targets.
CONCLUSIONS: Our results suggested that the mechanism of JWYHD in treating asthma might be related to limiting ILC2 responses. Our findings provided some pharmacological evidence for the clinical application of JWYHD in the treatment of asthma.
OBJECTIVE: To investigate the therapeutic effect as well as the underlying mechanisms of JWYHD on asthmatic mice.
METHODS: The ovalbumin (OVA)-induced mouse model was utilized, followed by the administration of JWYHD to allergic asthmatic mice. Subsequently, inflammatory cells in the bronchoalveolar lavage fluid (BALF) and lung tissues were conducted. The levels of various cytokines including interleukin (IL)-4, IL-5, IL-13, IL-33, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ in BALF, as well as the total immunoglobulin E (IgE) content in serum, were assessed. Lung function and tissue pathology examinations were performed to assess the protective impacts of JWYHD. The chemical components of JWYHD and its lung prototype compounds (referred to the chemical components present in JWYHD that were observed in the lung) were explored by ultra-high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS). RNA-seq analysis revealed the regulation mechanisms of JWYHD treating asthma. Furthermore, the effect of JWYHD on type 2 innate lymphoid cells (ILC2s) in asthmatic mice was detected by flow cytometry and Smart-RNA-seq analysis. Then molecular docking analysis was used to show the interaction between identified compounds and key targets.
RESULTS: JWYHD significantly attenuated the airway inflammation of asthmatic mice, reduced the levels of inflammatory cells in BALF, as well the levels of the cytokines IL-4, IL-5, IL-13, IL-33, and TNF-α in BALF and IgE in serum. Airway hyperresponsiveness (AHR) and lung inflammation infiltration were also alleviated by JWYHD. Moreover, RNA-seq analysis revealed that JWYHD attenuated airway inflammation in asthmatic mice via regulating immunity. Flow cytometry confirmed that JWYHD could inhibit ILC2 responses. ILC2 Smart-RNA-seq analysis showed that JWYHD impaired the inflammation reaction-related signaling pathways in ILC2s, and neuropilin-1 (Nrp1), endothelial transcription factor 3 (GATA3) and interleukin 1 receptor like protein 1 (ST2) might be the key targets. The molecular docking analysis investigating the connection between the primary targets and JWYHD\'s prototype compounds in the lung demonstrated that liquiritin apioside, icariin, glycyrrhizic acid, and uralsaponin B, identified through UPLC-Q-TOF/MS, exhibited significant affinity in binding to the mentioned key targets.
CONCLUSIONS: Our results suggested that the mechanism of JWYHD in treating asthma might be related to limiting ILC2 responses. Our findings provided some pharmacological evidence for the clinical application of JWYHD in the treatment of asthma.
摘要：
背景：加味阳和汤是加味阳和汤。YHD历史上用作解决慢性炎症的有效药物解决方案，在治疗哮喘方面具有很好的治疗潜力。然而,JWYHD对过敏性哮喘的作用机制尚不清楚.
目的：探讨JWYHD对哮喘小鼠的治疗作用及其机制。
方法：采用卵清蛋白（OVA）诱导的小鼠模型，然后对过敏性哮喘小鼠施用JWYHD。随后，支气管肺泡灌洗液（BALF）和肺组织中的炎症细胞。白细胞介素(IL)-4、IL-5、IL-13、IL-33、肿瘤坏死因子(TNF)-α、BALF中的干扰素（IFN）-γ，以及血清中总免疫球蛋白E（IgE）的含量，被评估。进行肺功能和组织病理学检查以评估JWYHD的保护性影响。通过超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF/MS)探索了JWYHD及其肺原型化合物的化学成分(指存在于JWYHD中的在肺中观察到的化学成分)。RNA-seq分析揭示了JWYHD治疗哮喘的调控机制。此外，通过流式细胞术和Smart-RNA-seq分析检测JWYHD对哮喘小鼠2型固有淋巴细胞(ILC2s)的影响.然后使用分子对接分析来显示鉴定的化合物与关键靶标之间的相互作用。
结果：JWYHD显著减轻哮喘小鼠的气道炎症，降低了BALF中炎症细胞的水平，以及血清中BALF和IgE中细胞因子IL-4，IL-5，IL-13，IL-33和TNF-α的水平。JWYHD还减轻了气道高反应性（AHR）和肺部炎症浸润。此外，RNA-seq分析显示JWYHD通过调节免疫减轻哮喘小鼠的气道炎症。流式细胞术证实JWYHD可以抑制ILC2反应。ILC2Smart-RNA-seq分析表明，JWYHD损害了ILC2s中炎症反应相关的信号通路，和神经纤毛蛋白-1（Nrp1），内皮转录因子3（GATA3）和白细胞介素1受体样蛋白1（ST2）可能是关键靶标。分子对接分析研究了主要靶标和JWYHD的原型化合物在肺中的联系，证明了甘草苷，淫羊藿苷,甘草酸，乌拉皂甙B,通过UPLC-Q-TOF/MS鉴定，在与上述关键靶标的结合中表现出显著的亲和力。
结论：我们的结果表明JWYHD治疗哮喘的机制可能与限制ILC2反应有关。我们的发现为JWYHD在哮喘治疗中的临床应用提供了一些药理学证据。
目的：探讨JWYHD对哮喘小鼠的治疗作用及其机制。
方法：采用卵清蛋白（OVA）诱导的小鼠模型，然后对过敏性哮喘小鼠施用JWYHD。随后，支气管肺泡灌洗液（BALF）和肺组织中的炎症细胞。白细胞介素(IL)-4、IL-5、IL-13、IL-33、肿瘤坏死因子(TNF)-α、BALF中的干扰素（IFN）-γ，以及血清中总免疫球蛋白E（IgE）的含量，被评估。进行肺功能和组织病理学检查以评估JWYHD的保护性影响。通过超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF/MS)探索了JWYHD及其肺原型化合物的化学成分(指存在于JWYHD中的在肺中观察到的化学成分)。RNA-seq分析揭示了JWYHD治疗哮喘的调控机制。此外，通过流式细胞术和Smart-RNA-seq分析检测JWYHD对哮喘小鼠2型固有淋巴细胞(ILC2s)的影响.然后使用分子对接分析来显示鉴定的化合物与关键靶标之间的相互作用。
结果：JWYHD显著减轻哮喘小鼠的气道炎症，降低了BALF中炎症细胞的水平，以及血清中BALF和IgE中细胞因子IL-4，IL-5，IL-13，IL-33和TNF-α的水平。JWYHD还减轻了气道高反应性（AHR）和肺部炎症浸润。此外，RNA-seq分析显示JWYHD通过调节免疫减轻哮喘小鼠的气道炎症。流式细胞术证实JWYHD可以抑制ILC2反应。ILC2Smart-RNA-seq分析表明，JWYHD损害了ILC2s中炎症反应相关的信号通路，和神经纤毛蛋白-1（Nrp1），内皮转录因子3（GATA3）和白细胞介素1受体样蛋白1（ST2）可能是关键靶标。分子对接分析研究了主要靶标和JWYHD的原型化合物在肺中的联系，证明了甘草苷，淫羊藿苷,甘草酸，乌拉皂甙B,通过UPLC-Q-TOF/MS鉴定，在与上述关键靶标的结合中表现出显著的亲和力。
结论：我们的结果表明JWYHD治疗哮喘的机制可能与限制ILC2反应有关。我们的发现为JWYHD在哮喘治疗中的临床应用提供了一些药理学证据。
