脾胃虚寒证代谢谱特征及姜枣暖九味颗粒作用机制研究

Metabolic profile research of the spleen-stomach vacuity cold pattern and the acting mechanism analysis of Jiang-zao-nuan-jiu-wei granules

[目的]通过基于小鼠血清超高液相色谱-四级杆飞行时间质谱联用技术的非靶标代谢组学探讨脾胃虚寒证的本质,以及姜枣暖九味颗粒的干预机制。[方法]SPF级BALB/c小鼠48只随机分为4个组,分别为正常对照组(对照组)、脾胃虚寒模型组(模型组)、脾胃虚寒模型+姜枣暖九味颗粒组(治疗组)、正常对照+姜枣暖九味颗粒组(对照治疗组)。模型组按0.1 mL/10 g灌胃4℃食醋一次;治疗组除造模外,每日另按照0.1 mL/10 g灌胃姜枣暖九味颗粒一次;对照治疗组只每天按照0.1 mL/10 g灌胃姜枣暖九味颗粒一次,持续10 d。实验期间内各组均自由摄食、饮水。记录各组小鼠每日进食量、饮水量;实验终点取小鼠血清,运用血清非靶标代谢组学对组间差异代谢物及代谢通路进行分析。统计学分析采用单因素方差分析和SNK-t检验。[结果]与对照组相比,模型组进食、饮水量下降;色氨酸代谢以及嘌呤代谢受到影响。与模型组相比,治疗组小鼠进食、饮水量增加,2组间的差异代谢物被富集在嘌呤代谢通路上。与对照组相比,对照治疗组小鼠进食、饮水量呈现降低趋势,同时血清D-谷氨酰胺升高。[结论]嘌呤代谢及色氨酸代谢异常可能为脾胃虚寒证的代谢谱特征。姜枣暖九味颗粒可通过纠正嘌呤代谢异常从而缓解脾胃虚寒证所致的临床表现,通过升高对照组小鼠D-谷氨酰胺以维护肠道黏膜的屏障功能。

[Objective]Research on the metabolic profile of the spleen-stomach vacuity cold pattern by the ultra high performance liquid tandem chromatography quadrupole time of flight mass spectrometry(UHPLC-QTOF-MS),and the effect mechanisms of the Jiang-zao-nuan-jiu-wei granules.[Methods]Forty-eight BALB/c mice in SPF grade were assigned randomly to 4 groups:control group,model group,model+treatment group,and control+treatment group.The model group was given 4℃vinegar,the model+treatment group was given Jiang-zao-nuan-jiu-wei granules besides vinegar,and the control+treatment group was only given the Jiang-zao-nuan-jiu-wei granules.All the groups by lavage administration with 0.1 mL/10 g/d for 10 days and all mice were given free access to water and commercial diets.The dietary and water intake for each mouse was recorded every day during the experimental period.After 24 hours of the last treatment,serum samples were collected from each mouse for the UHPLC-QTOF-MS analysis.One-way ANOVA and SNK-t test were used for statistical analysis.[Results]The dietary and water intake of model group were decreased compared to the control group,and the purine metabolism and tryptophan metabolism were differentially expressed pathways between the control and model group.Compared to the model group,the mice in the model+treatment group have higher dietary/water intake,and the purine metabolism is the only differential pathway was enriched between the two groups.While compared to the control group,the dietary/water intake of mice in the control+treatment group showed a decreased trend,and the D-Glutamine was increased.[Conclusion]Disorder of purine metabolism and tryptophan metabolism are the metabolic profile the spleen-stomach vacuity cold pattern.The Jiang-zao-nuan-jiu-wei granules relieve the symptoms of the pattern by attenuating the purine metabolism disorder.Furthermore,it benefits the control animal by increasing the D-glutamine.