尿液代谢组学探究葶苈大枣泻肺汤对哮喘大鼠的调节作用

Regulatory effect of Tingli Dazao Xiefei Decoction on asthmatic rats by urine metabolomics

基于超高效液相串联四极杆飞行时间质谱(UHPLC-Q-TOF-MS)的尿液代谢组学方法探究葶苈大枣泻肺汤(Tingli Dazao Xiefei Decoction, TLDZ)对过敏性哮喘大鼠的代谢调节机制。将SD雄性大鼠随机分为正常组、模型组、地塞米松组、TLDZ组,腹腔注射卵清白蛋白(ovalbumin, OVA)致敏联合雾化激发建立过敏性哮喘模型。通过UHPLC-Q-TOF-MS采集尿液样本代谢物,结合主成分分析(principal component analysis, PCA)建立各组大鼠的代谢轮廓,并通过正交偏最小二乘法判别分析(orthogonal partial least squares discriminant analysis, OPLS-DA)、t检验(P<0.05)和变量投影重要性(variable importance in the projection, VIP)>3筛选出模型组和TLDZ组间的差异代谢物;利用HMDB、METLIN等在线数据库进行代谢物的鉴定,通过MeV 4.8.0软件对标志物在各组的相对定量信息进行热图绘制和聚类分析。最后结合MetaboAnalyst、MBRole、KEGG数据库富集相关代谢通路并构建代谢网络。结果表明,TLDZ可从整体上有效调节哮喘大鼠尿液代谢轮廓的紊乱状态;结合多元统计分析和在线数据库,共筛选出45个模型组和TLDZ组间显著变化(P<0.05)的差异代谢物,主要富集到色氨酸代谢、组氨酸代谢、精氨酸和脯氨酸代谢等多个通路。TLDZ可通过调节相关的代谢通路,干预免疫稳态气道炎症等病理过程进而改善哮喘。该研究从尿液代谢组学的角度探究了TLDZ抗哮喘的分子机制,结合前期药理学研究,为TLDZ治疗哮喘的临床开发和应用提供科学依据。

Urine metabolomics based on ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(UHPLC-Q-TOF-MS)was utilized to investigate the metabolic regulation mechanism of Tingli Dazao Xiefei Decoction(TLDZ)in rats with allergic asthma.SD male rats were divided into a normal group,a model group,a dexamethasone group,and a TLDZ group.The allergic asthma model was established by intraperitoneal injection of ovalbumin(OVA)to induce allergy,combined with atomization excitation.Urine metabolites from all rats were collected by UHPLC-Q-TOF-MS.The metabolic profiles of rats in each group were built by principal component analysis(PCA).Besides,the differential metabolites between the model group and the TLDZ group were selected by orthogonal partial least squares discriminant analysis(OPLS-DA),t-test(P<0.05),and variable importance in the projection(VIP)values of more than 3.The differential metabolites were identified through HMDB,METLIN,and other online databases.Heat maps and clustering analysis for relative quantitative information of biomarkers in each group were drawn by MeV 4.8.0 software.Finally,MetaboAnalyst,MBRole,and KEGG databases were used to enrich related metabolic pathways and construct metabolic networks.The result demonstrated that TLDZ could effectively regulate the disordered urine metabolic profiles of asthmatic rats.Combined with multivariate statistical analysis and online databases,a total of 45 differential metabolites with significant changes(P<0.05)between the model group and the TLDZ group were screened out.Metabolic pathways including histidine metabolism,tryptophan metabolism,and arginine and proline metabolism were enriched.TLDZ could improve asthma by regulating related metabolic pathways and interfering with pathological processes such as immune homeostasis airway inflammation.The study investigates the molecular mechanism of anti-asthma of TLDZ from the perspective of urine metabolomics,and combined with previous pharmacological studies,it provides a scientific basis for the clinical development and application of TLDZ in the treatment of asthma.