Abstract：
BACKGROUND: Ephedra sinica Stapf (Mahuang) and Schisandra chinensis (Turcz.) Baill (Wuweizi) are commonly utilized in traditional Chinese medicine for the treatment of cough and asthma. The synergistic effect of Mahuang-Wuweizi herb pair enhances their efficacy in alleviating respiratory symptoms, making them extensively employed in the management of respiratory disorders. Although previous studies have demonstrated the therapeutic potential of Mahuang-Wuweizi in pulmonary fibrosis, the precise mechanism underlying their effectiveness against asthma remains elusive.
OBJECTIVE: The objective of this study is to investigate the mechanism underlying the preventive and therapeutic effects of Mahuang-Wuweizi herb pair on asthma progression, focusing on airway inflammation and airway remodeling.
METHODS: The active constituents and potential mechanisms of Mahuang-Wuweizi in the management of asthma were elucidated through network pharmacology analysis. Liquid chromatography tandem mass spectrometry (LC-MS/MS) was used to detect the main components of Mahuang-Wuweizi decoction. A rat model of bronchial asthma was established, and the effects of Mahuang-Wuweizi were investigated using hematoxylin-eosin (HE) staining, immunohistochemistry (IHC) staining, enzyme-linked immunosorbent assay (ELISA), Western blotting (WB), and real-time reverse transcription polymerase chain reaction (RT-qPCR).
RESULTS: The results of network pharmacological prediction showed that Mahuang had 22 active components and Wuweizi had 8 active components, with 225 potential targets. 1159 targets associated with asthma and 115 targets that overlap between drugs and diseases were identified. These include interleukin-6 (IL-6), tumor necrosis factor (TNF), Tumor Protein 53, interleukin-1β (IL-1β), as well as other essential targets. Additionally, there is a potential correlation between asthma and Phosphatidylinositol 3 kinase (PI3K)/Protein Kinase B (AKT) signaling pathway, calcium ion channels, nuclear factor-kappa B (NF-κB) signaling pathway, and other signaling pathways. The animal experiment results demonstrated that treatment with Mahuang and Wuweizi, in comparison to the model group, exhibited improvements in lung tissue pathological injury, reduction in collagen fiber accumulation around the airway and proliferation of airway smooth muscle, decrease in concentration levels of IL-6, TNF-α and IL-1β in lung tissue, as well as alleviation of airway inflammation. Furthermore, Mahuang and Wuweizi suppressed the expression of phospholipase C (PLC), transient receptor potential channel 1 (TRPC1), myosin light chain kinase (MLCK), NF-κB P65 protein in ovalbumin (OVA)-sensitized rat lung tissue and downregulated the mRNA expression of PLC, TRPC1, PI3K, AKT, NF-κB P65 in asthmatic rats. These findings were consistent with network pharmacological analysis.
CONCLUSIONS: The results show that the synergistic interaction between Mahuang and Wuweizi occur, and they can effectively reduce airway remodeling and airway inflammation induced by inhaling OVA in bronchial asthma rats by inhibiting the expression of PLC/TRPC1/PI3K/AKT/NF-κB signaling pathway. Therefore, Mahuang and Wuweizi may be potential drugs to treat asthma.
OBJECTIVE: The objective of this study is to investigate the mechanism underlying the preventive and therapeutic effects of Mahuang-Wuweizi herb pair on asthma progression, focusing on airway inflammation and airway remodeling.
METHODS: The active constituents and potential mechanisms of Mahuang-Wuweizi in the management of asthma were elucidated through network pharmacology analysis. Liquid chromatography tandem mass spectrometry (LC-MS/MS) was used to detect the main components of Mahuang-Wuweizi decoction. A rat model of bronchial asthma was established, and the effects of Mahuang-Wuweizi were investigated using hematoxylin-eosin (HE) staining, immunohistochemistry (IHC) staining, enzyme-linked immunosorbent assay (ELISA), Western blotting (WB), and real-time reverse transcription polymerase chain reaction (RT-qPCR).
RESULTS: The results of network pharmacological prediction showed that Mahuang had 22 active components and Wuweizi had 8 active components, with 225 potential targets. 1159 targets associated with asthma and 115 targets that overlap between drugs and diseases were identified. These include interleukin-6 (IL-6), tumor necrosis factor (TNF), Tumor Protein 53, interleukin-1β (IL-1β), as well as other essential targets. Additionally, there is a potential correlation between asthma and Phosphatidylinositol 3 kinase (PI3K)/Protein Kinase B (AKT) signaling pathway, calcium ion channels, nuclear factor-kappa B (NF-κB) signaling pathway, and other signaling pathways. The animal experiment results demonstrated that treatment with Mahuang and Wuweizi, in comparison to the model group, exhibited improvements in lung tissue pathological injury, reduction in collagen fiber accumulation around the airway and proliferation of airway smooth muscle, decrease in concentration levels of IL-6, TNF-α and IL-1β in lung tissue, as well as alleviation of airway inflammation. Furthermore, Mahuang and Wuweizi suppressed the expression of phospholipase C (PLC), transient receptor potential channel 1 (TRPC1), myosin light chain kinase (MLCK), NF-κB P65 protein in ovalbumin (OVA)-sensitized rat lung tissue and downregulated the mRNA expression of PLC, TRPC1, PI3K, AKT, NF-κB P65 in asthmatic rats. These findings were consistent with network pharmacological analysis.
CONCLUSIONS: The results show that the synergistic interaction between Mahuang and Wuweizi occur, and they can effectively reduce airway remodeling and airway inflammation induced by inhaling OVA in bronchial asthma rats by inhibiting the expression of PLC/TRPC1/PI3K/AKT/NF-κB signaling pathway. Therefore, Mahuang and Wuweizi may be potential drugs to treat asthma.
摘要：
背景：麻黄（麻黄）和五味子（Turcz。）Baill（五味子）常用于中医治疗咳嗽和哮喘。麻黄-五味子中药对的协同作用增强了其缓解呼吸道症状的功效，使它们广泛用于呼吸系统疾病的管理。尽管先前的研究已经证明了麻黄-五味子在肺纤维化中的治疗潜力，它们对哮喘有效的确切机制仍然难以捉摸.
目的：本研究的目的是探讨麻黄-五味子对哮喘进展的预防和治疗作用的潜在机制。关注气道炎症和气道重塑。
方法：通过网络药理学分析，阐明麻黄五味子治疗哮喘的有效成分和潜在作用机制。采用液相色谱-串联质谱(LC-MS/MS)检测麻黄五味子汤的主要成分。建立支气管哮喘大鼠模型,并利用苏木精-伊红(HE)染色研究了麻黄-五味子的作用,免疫组化（IHC）染色，酶联免疫吸附测定(ELISA),蛋白质印迹(WB),和实时逆转录聚合酶链反应（RT-qPCR）。
结果：网络药理预测结果表明，麻黄有22种活性成分，五味子有8种活性成分，有225个潜在目标。确定了1159个与哮喘相关的靶标和115个在药物和疾病之间重叠的靶标。这些包括白细胞介素-6（IL-6），肿瘤坏死因子(TNF),肿瘤蛋白53,白细胞介素-1β(IL-1β),以及其他重要目标。此外,磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)信号通路与哮喘之间存在潜在的相关性,钙离子通道，核因子-κB(NF-κB)信号通路,和其他信号通路。动物实验结果表明，麻黄和五味子处理，与模型组相比，表现出改善肺组织病理损伤，减少气道周围胶原纤维的积累和气道平滑肌的增殖，肺组织中IL-6，TNF-α和IL-1β的浓度水平降低，以及缓解气道炎症。此外，麻黄和五味子抑制磷脂酶C（PLC）的表达，瞬时受体电位通道1（TRPC1），肌球蛋白轻链激酶（MLCK），卵清蛋白致敏大鼠肺组织中NF-κBP65蛋白表达下调,TRPC1,PI3K,AKT,NF-κBP65在哮喘大鼠中的表达.这些发现与网络药理学分析一致。
结论：结果表明，麻黄与五味子之间存在协同作用，通过抑制PLC/TRPC1/PI3K/AKT/NF-κB信号通路的表达,有效减轻OVA吸入诱导的支气管哮喘大鼠气道重塑和气道炎症反应。因此,麻黄和五味子可能是治疗哮喘的潜在药物。
目的：本研究的目的是探讨麻黄-五味子对哮喘进展的预防和治疗作用的潜在机制。关注气道炎症和气道重塑。
方法：通过网络药理学分析，阐明麻黄五味子治疗哮喘的有效成分和潜在作用机制。采用液相色谱-串联质谱(LC-MS/MS)检测麻黄五味子汤的主要成分。建立支气管哮喘大鼠模型,并利用苏木精-伊红(HE)染色研究了麻黄-五味子的作用,免疫组化（IHC）染色，酶联免疫吸附测定(ELISA),蛋白质印迹(WB),和实时逆转录聚合酶链反应（RT-qPCR）。
结果：网络药理预测结果表明，麻黄有22种活性成分，五味子有8种活性成分，有225个潜在目标。确定了1159个与哮喘相关的靶标和115个在药物和疾病之间重叠的靶标。这些包括白细胞介素-6（IL-6），肿瘤坏死因子(TNF),肿瘤蛋白53,白细胞介素-1β(IL-1β),以及其他重要目标。此外,磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)信号通路与哮喘之间存在潜在的相关性,钙离子通道，核因子-κB(NF-κB)信号通路,和其他信号通路。动物实验结果表明，麻黄和五味子处理，与模型组相比，表现出改善肺组织病理损伤，减少气道周围胶原纤维的积累和气道平滑肌的增殖，肺组织中IL-6，TNF-α和IL-1β的浓度水平降低，以及缓解气道炎症。此外，麻黄和五味子抑制磷脂酶C（PLC）的表达，瞬时受体电位通道1（TRPC1），肌球蛋白轻链激酶（MLCK），卵清蛋白致敏大鼠肺组织中NF-κBP65蛋白表达下调,TRPC1,PI3K,AKT,NF-κBP65在哮喘大鼠中的表达.这些发现与网络药理学分析一致。
结论：结果表明，麻黄与五味子之间存在协同作用，通过抑制PLC/TRPC1/PI3K/AKT/NF-κB信号通路的表达,有效减轻OVA吸入诱导的支气管哮喘大鼠气道重塑和气道炎症反应。因此,麻黄和五味子可能是治疗哮喘的潜在药物。
