基于UPLC-QTOF-MS技术的桔梗叶提取物对小鼠抑郁作用的尿液代谢组研究

Study on urine metabolome of platycodon grandiflorus leaves extracts on depression in mice based on UPLC-QTOF-MS technology

关键词:  桔梗叶  抑郁症  代谢组学  代谢通路

Key words:  platycodon grandiflorum leaves  depression  metabolomics  metabolic pathway

摘要:

[目的] 基于超高效液相色谱-飞行时间质谱（UPLC-QTOF-MS）技术对桔梗叶提取物干预小鼠抑郁症模型进行研究，探讨抑郁症小鼠的发病机制及桔梗叶对其干预作用的作用机制。[方法] 采用腹腔注射脂多糖方法诱导小鼠抑郁症模型（1 mg/kg），按照体质量随机分为模型组和给药组。另选8只同批次的健康小鼠作为空白组。给药组灌胃给与桔梗叶进行干预[400 mg/（kg·d）]，连续1周，空白组和模型组给等体积的生理盐水。采用白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）进行评价，同时采用尿液代谢组学技术综合评价桔梗叶的药效及作用机制。[结果] 生化指标结果表明，与空白组相比，模型组小鼠前额皮质IL-6、TNF-α表达明显升高（P<0.01）；与模型组相比，给药组各项指标具有显著回调趋势（P<0.05）。尿液代谢组学研究发现，3组小鼠代谢轮廓聚类明显，且给药组更趋近于空白组的代谢分布，证明在整体代谢水平上，给药组的代谢更趋近于健康小鼠状态。通过Progenesis QI软件深度挖掘，对比数据库鉴定出29个尿液生物标记物，涉及亚油酸代谢、花生四烯酸代谢、谷胱甘肽代谢、鞘脂代谢、能量代谢等，并给予桔梗叶后上述代谢通路均出现回调趋势。[结论] 桔梗叶提取物对抑郁症小鼠有一定程度的治疗作用。多成分多径路的调节方式可能是其有效性的关键，通过调节亚油酸代谢、花生三烯酸代谢等机制间接达到治疗抑郁症的目的。

Abstract:

[Objective] To investigate the effect of platycodon grandiflorum leaves extract on depression model in mice based on UPLC-QTOF-MS technology,and to explore the pathogenesis of depression in mice and the mechanism of platycodon grandiflorum leaves intervention.[Methods] The depression model of mice was induced by intraperitoneal injection of lipopolysaccharide (1 mg/kg). The mice were randomly divided into model group and drug group according to their body weight. Eight healthy mice of the same batch were selected as the control group. The mice in the administration group were treated with platycodon grandiflorum leaves by gavage[400 mg/(kg·d)] for one week. The control group and the model group were treated with physiological saline of equal volume. IL-6 and TNF-α were used for evaluation,and urine metabolomics was used for data acquisition and data processing. By tracing the key metabolic enzymes and pathways upstream through terminal metabolites to evaluated comprehensively from the overall metabolic level the pharmacodynamics and mechanism of platycodon grandiflorum leaves.[Results] The results of biochemical indexes showed that the expressions of IL-6 and TNF-α in the prefrontal cortex of the model group were significantly higher than those in the control group(P<0.01);compared with the model group,the indexes in the drug group showed a significant retrograde trend(P<0.05). The results of urine metabolomics study showed that the metabolic profiles of three groups of mice clustered significantly. Moreover,the metabolic distribution of the drug group was closer to that of the control group,which showed that the metabolic status of the drug group was closer to that of the healthy mice at the overall metabolic level. Progenesis QI software was used to identify 29 urine biomarkers which contributed significantly to grouping. It involves Linoleic acid metabolism,Arachidonic acid metabolism,Glutathione metabolism,Sphingolipid metabolism,Energy metabolism and so on. The above metabolic pathways showed a tendency to be reversed after platycodon grandiflorum leaves were given.[Conclusion] The extract of platycodon grandiflorum leaves has a certain therapeutic effect on depressive mice. The regulation of multi-component and multi-path pathway may be the key to its effectiveness. It can indirectly intervene depression by regulating Linoleic acid metabolism and Arachidonic acid metabolism.