背景:柴胡是柴胡的根。(BC)和经典芳香中药。柴胡的传统药理作用是缓解支气管痉挛,扩张气道,促进呼吸道炎症的消退,从而减少哮喘症状。
目的:研究已经证明了BC水提取物在哮喘治疗中的功效。然而,挥发油的潜在作用,BC中的另一种活性成分,哮喘仍未被探索。值得注意的是,挥发油以其易于吸收和直接瞄准受影响地区而闻名,在减轻气道炎症方面具有明显的优势。本研究旨在通过体内外药理实验来阐明BC油的平喘作用机制。
方法:首先,利用OVA诱导的SD大鼠哮喘模型,通过肺功能监测评价BC油的药理作用,HE染色,流式细胞术,ELISA,和RT-qPCR。通过结合来自大鼠模型的肺组织和来自公共数据库的气道平滑肌组织的转录组学分析,进一步分析了抗哮喘的机制。最初,GC-MS用于分析BC油的成分。在16-HBE中评估了抗哮喘活性,RBL-2H3和ASMC细胞使用CAMKII抑制剂探索BC-油调节的关键信号转导。此外,利用分子对接和钙流测定法筛选和鉴定BC油中的活性成分。
结果:口服BC油通过降低气道阻力和弹性阻力显著增强哮喘SD大鼠的肺功能。此外,BC-油抑制炎症细胞因子,包括血清IL-2,肺Il1b,Tnf,和Cxcl13,证明了有效的抗炎和免疫调节作用。在这项研究中,我们使用公开转录组数据分析了OR2W3在哮喘中的重要作用.此外,我们表明BC油调节大鼠肺组织Olr1433和GNAL的表达。BC油减少RBL-2H3细胞中的脱颗粒并抑制Il3和Tnf的基因表达,并抑制16-HBE细胞中IL8和TNF的基因表达。BC-油还减弱气道平滑肌细胞增殖和Acta2和Ccnd1的表达。此外,BC油通过激活CAMKII调节哮喘相关的细胞过程。GC-MS分析确定了BC油的11种成分,和正十六烷酸,通过分子对接,鉴定了来自BC油的亚油酸和油酸与OR2W3相互作用。钙流测定显示亚油酸是OR2W3的重要激活剂,并表明BC油通过异位嗅觉信号通路缓解哮喘。
结论:在分子和细胞水平上揭示了BC油通过OR2W3信号转导治疗哮喘的机制。
BACKGROUND: Radix Bupleuri is the root of Bupleurum chinense DC. (BC) and a classic aromatic traditional Chinese medicine. The traditional pharmacological effects of Radix Bupleuri are alleviating bronchial spasms, dilating airways, and promoting the resolution of respiratory inflammation, thereby reducing asthma symptoms.
OBJECTIVE: Studies have demonstrated the efficacy of water extracts from BC in asthma treatment. However, the potential role of volatile oil, another active constituent in BC, remains unexplored with asthma. Notably, volatile oil is renowned for its ease of absorption and direct targeting of affected areas, offering distinct advantages in alleviating airway inflammation. This study aims to explain the anti-asthmatic mechanism of BC-oil through in vivo and in vitro pharmacological experiments.
METHODS: Firstly, the OVA-induced SD rat asthma model was utilized to evaluate the pharmacological effect of BC-oil by lung function monitoring, HE staining, flow cytometry, ELISA, and RT-qPCR. The anti-asthmatic mechanism was further analyzed by combining transcriptomic analysis of lung tissue from rat model and airway smooth muscle tissue from public database. Initially, GC-MS was used to analyze the components of BC-oil. The anti-asthmatic activity was evaluated in 16-HBE, RBL-2H3, and ASMC cells using CAMKII inhibitors to explore of the critical signal transduction regulated by BC-oil. Furthermore, molecular docking and calcium flow assay were utilized to screen and identify the active components from BC-oil.
RESULTS: Oral administration of BC-oil significantly enhanced pulmonary function in asthmatic SD rats by reducing airway resistance and elastic resistance. Additionally, BC-oil inhibited inflammatory cytokines, including serum IL-2, pulmonary Il1b, Tnf, and Cxcl13, demonstrating potent anti-inflammatory and immunomodulatory effects. In this study, we analyzed the significant role of OR2W3 in asthma using public transcriptomic data. Furthermore, we indicated that BC-oil regulated the expression of Olr1433 and GNAL in rat lung tissue. BC-oil reduced degranulation and inhibited gene expression of Il3 and Tnf in RBL-2H3 cells and suppressed gene expression of IL8 and TNF in 16-HBE cells. BC-oil also attenuated airway smooth muscle cell proliferation and expression of Acta2 and Ccnd1. Furthermore, BC-oil regulates asthma-related cellular processes by activating CAMKII. GC-MS analysis identified 11 components of BC-oil, and n-hexadecanoic acid, linoleic acid and oleic acid from BC-oil were identified to interact with OR2W3 by molecular docking. The calcium flow assay revealed linoleic acid as a significant activator of OR2W3 and indicated that BC-oil alleviated asthma through the ectopic olfactory signaling pathway.
CONCLUSIONS: The mechanism of BC-oil in treating asthma through signal transduction of OR2W3 is revealed at the molecular and cellular levels.