铁玘美, 曾梦楠, 张贝贝, 陈艳玲, 肖超远, 陈霖, 康珂瑞, 靳佳音, 冯卫生, 郑晓珂. 基于代谢组学分析茯苓中过氧去氢土莫酸对哮喘气道炎症的干预作用J. 药学学报, 2026, 61(1): 222-232. DOI: 10.16438/j.0513-4870.2025-0560
引用本文: 铁玘美, 曾梦楠, 张贝贝, 陈艳玲, 肖超远, 陈霖, 康珂瑞, 靳佳音, 冯卫生, 郑晓珂. 基于代谢组学分析茯苓中过氧去氢土莫酸对哮喘气道炎症的干预作用J. 药学学报, 2026, 61(1): 222-232. DOI: 10.16438/j.0513-4870.2025-0560
TIE Qi-mei, ZENG Meng-nan, ZHANG Bei-bei, CHEN Yan-ling, XIAO Chao-yuan, CHEN Lin, KANG Ke-rui, JIN Jia-yin, FENG Wei-sheng, ZHENG Xiao-ke. Metabolomics-based analysis of the interventional effects of peroxydehydrotumulosic acid in Poria cocos on airway inflammation in asthmaJ. Acta Pharmaceutica Sinica, 2026, 61(1): 222-232. DOI: 10.16438/j.0513-4870.2025-0560
Citation: TIE Qi-mei, ZENG Meng-nan, ZHANG Bei-bei, CHEN Yan-ling, XIAO Chao-yuan, CHEN Lin, KANG Ke-rui, JIN Jia-yin, FENG Wei-sheng, ZHENG Xiao-ke. Metabolomics-based analysis of the interventional effects of peroxydehydrotumulosic acid in Poria cocos on airway inflammation in asthmaJ. Acta Pharmaceutica Sinica, 2026, 61(1): 222-232. DOI: 10.16438/j.0513-4870.2025-0560

基于代谢组学分析茯苓中过氧去氢土莫酸对哮喘气道炎症的干预作用

Metabolomics-based analysis of the interventional effects of peroxydehydrotumulosic acid in Poria cocos on airway inflammation in asthma

  • 摘要: 探究茯苓中过氧去氢土莫酸(peroxydehydrotumulosic acid, PHA) 对过敏性哮喘小鼠的干预作用, 并基于代谢组学探究其作用机制。雄性BALB/c小鼠随机分为正常组(CON)、模型组(M)、阳性药孟鲁斯特钠组(Y, 1.3 mg·kg-1)、PHA低剂量组(PHA-L, 15 mg·kg-1) 和PHA高剂量组(PHA-H, 30 mg·kg-1)。采用腹腔注射卵清蛋白(ovalbumin, OVA) 和氢氧化铝佐剂进行致敏, OVA雾化激发建立过敏性哮喘小鼠模型。造模前30 min灌胃给予相应药物, 连续给药7天, 最后一次给药24 h后检测小鼠咳喘指标、肺功能、肺系数, 苏木精-伊红(hematoxylin-eosin, HE) 染色观察小鼠肺组织病理变化, 马松(Masson) 染色观察其胶原纤维沉积水平, 酶联免疫吸附(ELISA) 测定小鼠血清和肺泡灌洗液中炎症因子水平, 流式细胞术检测小鼠肺组织凋亡细胞水平以及血中Th17、Tregs细胞水平, 采用液质联用技术(LC-MS), 对不同处理组小鼠的血清样本小分子代谢产物进行分析, 运用主成分分析(principal component analysis, PCA) 与偏最小二乘判别分析(orthogonal partial least squares discriminant analysis, OPLS-DA) 等多元统计方法, 筛选差异代谢物并通过代谢通路富集分析, 探究特征代谢物所参与的生物代谢途径。结果显示, 与M组相比, PHA组可以显著改善小鼠的肺功能, 降低肺系数, 减轻肺组织病理损伤和胶原纤维沉积, 显著下调血清中白介素4 (interleukin 4, IL4)、免疫球蛋白E (immunoglobulin E, IgE)、白介素17A (interleukin 17A, IL17A) 以及肺泡灌洗液中肿瘤坏死因子α (tumor necrosis factor-α, TNF-α) 的水平, 抑制肺组织细胞凋亡以及Th17细胞的生成, 增加Tregs细胞水平。代谢组学分析M组与PHA组内源性物质分离明显, 筛选出差异代谢物63个, 标志性差异代谢物11个, 主要涉及花生四烯酸代谢, 视黄醇代谢, 苯丙氨酸、色氨酸、酪氨酸的生物合成, 甘油磷脂代谢和酪氨酸代谢。PHA可以通过抑制炎症因子IL4、IL17A的产生, 阻碍炎症细胞的招募, 调节部分差异代谢物及代谢通路紊乱, 从而改善哮喘的小鼠的病理状态。动物实验获得河南中医药大学动物伦理委员会批准和监督(伦理编号: DWLL2018080003)。

     

    Abstract: The study aims to investigate the interventional effect of peroxydehydrotumulosic acid (PHA) in Poria cocos on allergic asthma mice, and to investigate its mechanism of action based on metabolomics. Male BALB/c mice were randomly divided into normal group (CON), model group (M), positive drug montelukastna group (Y, 1.3 mg·kg-1), PHA low-dose group (PHA-L, 15 mg·kg-1), and PHA high-dose group (PHA-H, 30 mg·kg-1). The mouse model of allergic asthma was established by intraperitoneal injection of ovalbumin (OVA) and aluminium hydroxide adjuvant for sensitisation and OVA nebulisation for excitation. The corresponding drugs were given by gavage 30 min before modelling, and the drugs were administered for 7 consecutive days. 24 h after the last administration of the drugs, the mice were tested for cough and asthma indexes, lung function and lung coefficient, the mice were stained with hematoxylin-eosin (HE) to observe the pathological changes of the lung histology, and the mice were stained with Masson to observe the level of collagen fibre deposition, and the levels of inflammatory factors in the serum and alveolar lavage fluid of the mice were measured by enzyme-linked immunosorbent assay (ELISA) and flow cytometry to determine the levels of inflammatory factors in the mice. Apoptotic cell levels in lung tissue and Th17 and Tregs cell levels in blood were detected by flow cytometry, and small molecule metabolites were analysed by liquid-liquid mass spectrometry (LC-MS) in serum samples of mice of different treatment groups. The metabolic pathway enrichment analysis was used to investigate the biometabolic pathways involved in the characterised metabolites. The results indicated a comparison with group M, PHA significantly improved lung function, reduced lung coefficient, attenuated lung histopathological damage and collagen fibre deposition, significantly down-regulated the levels of interleukin 4 (IL4), immunoglobulin E (IgE), interleukin 17A (IL17A) in blood and tumor necrosis factor-α (TNF-α) in alveolar lavage fluid, inhibited apoptosis and the generation of Th17 cells, and increased the level of Tregs cells in lung tissue. Metabolomics analysis of endogenous substances in the M and PHA groups separated significantly, and 63 differential metabolites were screened, mainly involving arachidonic acid metabolism, retinol metabolism, biosynthesis of phenylalanine, tryptophan, and tyrosine, glycerophospholipid metabolism, and tyrosine metabolism. PHA can improve the pathological state of mice with asthma by inhibiting the production of inflammatory factors IL4 and IL17A, hindering the recruitment of inflammatory cells, and regulating some differential metabolites and metabolic pathway disorders. The animal experiment was approved and supervised by the Animal Ethics Committee of Henan University of Traditional Chinese Medicine (ethics number: DWLL2018080003).

     

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