基于粪便代谢物组学分析麻黄细辛附子汤干预H1N1流感病毒感染小鼠的作用机制

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目的:运用代谢组学方法研究麻黄细辛附子汤(Mahuang Xixin Fuzi Tang,MXF)干预H1N1流感病毒感染小鼠粪便样品中内源性物质的变化,寻找与疾病相关的生物标志物,探讨MXF干预H1N1流感病毒感染可能的作用机制。方法:将30只KM小鼠随机等分为空白组、模型组和MXF组,小鼠滴鼻感染H1N1流感病毒后灌服MXF(给药剂量20 m L·kg~(-1)),模型组和空白组灌服等量水,连续给药7 d并测量各组小鼠的体重和肛温,收集各组小鼠的粪便。采用Halo C_(18)色谱柱(2.1 mm×100 mm,2.7μm),流动相0.05%甲酸水溶液-0.05%甲酸乙腈溶液梯度洗脱,质谱扫描范围m/z 80~1 200,对粪便样品进行LC-MS测定并结合主成分分析和偏最小二乘判别分析寻找潜在生物标志物。结果:空白组、模型组和MXF组代谢模式显著不同,鉴定出了9个潜在生物标志物,分别为磷脂酰乙醇胺、磷脂酰丝氨酸、磷脂酰胆碱、溶血磷脂酰胆碱、吡哆醛、草酰乙酸、琥珀酸、褪黑素和L-犬尿氨酸。结论:MXF干预H1N1流感病毒感染的机制可能与其对色氨酸代谢、维生素B6代谢、甘油磷脂代谢和三羧酸循环等代谢紊乱的回调作用有一定相关性。 OBJECTIVE: To study the changes of endogenous substances in fecal samples of mice infected with H1N1 influenza virus by Mahuang Xixin Fuzi Tang (MXF) by means of metabonomics to find disease-related biomarkers and explore the effects of MXF intervention Possible Mechanism of H1N1 Influenza Virus Infection. Methods: Thirty KM mice were randomly divided into blank group, model group and MXF group. Mice were intranasally infected with the H1N1 influenza virus (MXF) and the dose of MXF (20 m L · kg -1) The rats in the two groups were fed with the same amount of water for 7 days. The body weight and rectal temperature of the mice in each group were measured, and the feces of each group were collected. The mobile phase was eluted with 0.05% formic acid-0.05% formic acid in acetonitrile by gradient elution on a Halo C_ (18) column (2.1 mm × 100 mm, 2.7 μm) LC-MS determination combined with principal component analysis and partial least-squares discriminant analysis to find potential biomarkers. Results: The metabolic patterns of blank group, model group and MXF group were significantly different. Nine potential biomarkers were identified as phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine, lysophosphatidylcholine, pyridoxal, Oxaloacetate, succinate, melatonin and L-kynurenine. CONCLUSION: The mechanism of MXF in preventing H1N1 influenza virus infection may be related to its effect on the metabolic regulation of tryptophan metabolism, vitamin B6 metabolism, glycerophospholipid metabolism and tricarboxylic acid cycle.
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