目的 基于超高液相色谱-四极杆静电场轨道阱质谱法(ultra-high performance liquid chromatography-qua-drupole electrostatic field orbital trap mass spectrometry,UHPLC-QE-MS)研究达原饮对模型小鼠的干预及肺组织代谢物的影响,以探究达原饮治疗呼吸道合胞病毒(Respiratory syncytial virus,RSV)寒湿郁肺证的作用机制。方法 采用寒湿刺激加RSV滴鼻感染的方法建立RSV寒湿郁肺证小鼠模型,综合小鼠活动状态、肺指数和肺组织病理,结合肺组织中白介素-6(Interleukin-6,IL-6)和白介素-1 β(Interleukin-1β,IL-1 β)炎症因子的含量,初步评价达原饮对RSV寒湿郁肺证小鼠的治疗效果。利用UHPLC-QE-MS技术检测正常组、模型组、达原饮中剂量组小鼠肺组织代谢物,以正交偏最小二判别分析进行多元统计分析模式识别,以P<0。05且VIP值>1为条件筛选差异代谢物,并借助MetaboAnalyst 5。0数据库富集代谢通路。结果 达原饮可以显著改善RSV寒湿郁肺证小鼠的一般状态,降低肺指数,改善肺组织病理,并降低肺组织中IL-6、IL-1β的含量。代谢组学结果显示,与达原饮调控相关的差异代谢物共有35个,主要涉及嘌呤代谢、花生四烯酸、甘氨酸、丝氨酸和苏氨酸代谢等代谢通路。结论 从代谢组学角度分析了达原饮治疗RSV寒湿郁肺证的作用机制,为达原饮抗病毒研究及临床应用奠定了基础。
Mechanism of Famous Classical Formula Dayuanyin Decoction(达原饮)in Treatment of Mice Model with RSV and Cold-Dampness in Lung Syndrome Based on Metabonomics
Objective Ultra-high performance liquid chromatography-quadrupole electrostatic field orbital trap mass spec-trometry(UHPLC-QE-MS)was employed in this study to observe the effect of Dayuanyin Decoction(达原饮)on the lung tis-sues metabolites of model mice and reveal the mechanism of Dayuanyin Decoction in the treatment of respiratory syncytial virus(RSV)with cold-dampness in lung syndrome.Methods The mice were placed in a cold-dampness environment and infected with RSV to establish the mouse model.The therapeutic effect of Dayuanyin Decoction on RSV with cold-dampness in lung syn-drome was preliminarily evaluated based on the mice activity status,lung index,lung tissue morphology,and the content of inflam-matory cytokines such as interleukin-6(IL-6)and interleukin-1β(IL-1β).UHPLC-QE-MS was employed to profile the endogenous metabolites of lung tissues samples in normal group,model group and medium-dose Dayuanyin Decoction group.Or-thogonal partial least squares-discriminant analysis(OPLS-DA)was performed for multivariate analyses pattern recognition.Potential biomarkers were screened with P<0.05 and VIP>1,and the metabolic pathways were enriched by MetaboAnalyst 5.0.Results Dayuanyin Decoction significantly improved the general state and lung tissue morphology of mice with RSV and cold-dampness in lung syndrome,reduced the lung index and lowered the levels of IL-6 and IL-1 β in lung.A total of 35 potential biomarkers were predicted to be related to the regulatory effect of Dayuanyin Decoction,which were mainly involved in purine me-tabolism,arachidonic acid metabolism,glycine,serine and threonine metabolism.Conclusion This study employed metabolomics to analyze the mechanism of Dayuanyin Decoction in the treatment of RSV with cold-dampness in lung syndrome,forming a foundation for the antiviral research and clinical application of Dayuanyin Decoction.
Dayuanyin Decoction(达原饮)respiratory syncytial virusmetabonomicscold-dampness in lung syndrome