基于UPLC-QTOF-MS技术的藿香正气口服液治疗大鼠湿困脾胃证的血清代谢组学研究

Studies on the Therapeutic Effects of Huoxiang Zhengqi Oral Liquid on Dampness Obstructing Spleen-stomach Syndrome in Rats Using UPLC-QTOF-MS-based Serum Metabonomics

目的基于超高效液相色谱-四极杆-飞行时间质谱联用(UPLC-QTOF-MS)技术的血清代谢组学方法寻找与湿困脾胃证相关的潜在生物标志物,探讨藿香正气口服液对湿困脾胃证的干预机制。方法采用综合物理造模方法建立湿困脾胃证大鼠模型。观测大鼠体质量、体长、尾长、腹围指数与自主活动次数;利用UPLCQTOF-MS方法对大鼠血清进行代谢组分析,结合正交偏最小二乘法-判别分析(OPLS-DA),寻找藿香正气口服液治疗湿困脾胃证大鼠后相关潜在生物标志物的变化及其相关代谢通路。结果与正常组比较,模型组大鼠的体质量、体长、自主活动次数明显减少(P<0.001),腹围指数明显升高(P<0.001);正离子和负离子模式下分别鉴定了与湿困脾胃证相关的15、10个血清潜在生物标志物,涉及磷脂代谢、氨基酸代谢、能量代谢等9条代谢通路。藿香正气口服液对湿困脾胃证模型大鼠的血清代谢表型能产生有效的干预,并使14个潜在生物标志物产生显著回调(P<0.05)。结论藿香正气口服液对大鼠湿困脾胃证的干预作用可能与调节磷脂代谢、氨基酸代谢、能量代谢等通路有关。

Objective To identify the potential biomarkers related to the dampness obstructing spleen-stomach syndrome(DOSS) through a serum metabolomics method based on Ultra High Performance Liquid Tandem Chromatography Quadrupole Time of Flight Mass Spectrometry(UPLC-QTOF-MS),and illustrate the intervention mechanism of Huoxiang Zhengqi oral liquid(HZOL).Methods Comprehensive physical methods were applied to establish the rat DOSS model.Body mass,body length,tail length,abdominal girth index,and locomotor activity were observed.UPLC-QTOF-MS combined with orthogonal partial least squares-discriminant analysis(OPLS-DA)were used to analyze the metabonomics of rat serum and to identify potential biomarkers and related metabolic pathways in rats with DOSS after administration of HZOL.Results Compared with the healthy control group,rats with DOSS showed body mass loss(P<0.001),slower growth of body length(P<0.001),increased abdominal girth index(P<0.001),and significantly decreased locomotor activity(P<0.001).15 and 10 potential biomarkers associated with DOSS in rat serum were identified under the positive ion and negative ion modes,respectively,which involved 9 metabolic pathways such as phospholipid metabolism,amino acid metabolism and energy metabolism.HZOL effectively intervened in the serum metabolic phenotype of the model rats,and significantly regulated 14 potential biomarkers(P<0.05).Conclusion The mechanism of HZOL intervention on the DOSS in rats may relate to the regulation of phospholipid metabolism,amino acid metabolism,energy metabolism,and so on.