基于~1H-NMR代谢组学的酸枣仁改善失眠大鼠睡眠作用机制研究

~1H-NMR based metabolomic study of sedative and hypnotic effect of Ziziphi Spinosae Semen in rats

目的利用氢核磁共振(~1H-NMR)代谢组学技术研究酸枣仁对对氯苯丙氨酸(PCPA)诱导的失眠大鼠血清内源性代谢物的影响,探讨酸枣仁改善睡眠的作用机制。方法大鼠ipPCPA溶液(400mg/kg)制备失眠模型,采用1H-NMR代谢组学技术,以主成分分析(PCA)、偏最小二乘法判别分析(PLS-DA)及正交偏最小二乘法判别分析(OPLS-DA)等多元统计分析方法为手段,对各组大鼠血清代谢物进行分析,筛选出潜在生物标志物;借助MatPA分析构建潜在靶标代谢通路和代谢网络。结果 1H-NMR代谢组学分析方法共筛选11种与失眠相关的潜在生物标志物;通过MatPA分析得到8条潜在靶标代谢通路,包括谷氨酸和谷氨酰胺代谢,牛磺酸和亚牛磺酸代谢,丙氨酸、天冬氨酸和谷氨酸的代谢,乙醛酸和二羧酸代谢,甘氨酸、丝氨酸和苏氨酸代谢,丙酮酸代谢,色氨酸代谢,三羧酸(TCA)循环。酸枣仁中剂量(15 g/kg)和高剂量(30 g/kg)均可显著回调11种潜在生物标志物。结论酸枣仁可通过调控失眠大鼠的能量代谢和氨基酸代谢,使大鼠内源性代谢物趋近正常水平,为进一步阐明酸枣仁改善睡眠的作用机制提供依据。

Objective ^1H-NMR based metabolomic approach has been employed to study the intervention effects of Ziziphi Spinosae Semen(ZSS) on p-chlorophenylalanine(PCPA)-induced insomnia using a rat model. Methods 1 H-NMR analysis was used to identify the various compounds of ZSS, and the quantitative data were further processed by principal component analysis(PCA), partial least squares discrimination analysis(PLS-DA), and orthogonality PLS-DA to find potential biomarkers. In addition, MetPA analysis was used for pathway analysis. Results Multivariate analysis revealed that 11 perturbed endogenous metabolites in the serum could be reversed by ZSS at dosage of 15 g/kg and 30 g/kg, and MetPA analysis revealed that the anti-hypnotic effect of ZSS was probably related with regulation of glutamate and glutamite metabolism, taurine and taurine metabolism, alanine, aspartic acid and glutamate metabolism, glyoxylic acid, and dicarboxylic acid metabolism, glycine, serine and threonine metabolism, pyruvate metabolism,tryptophan metabolism, and tricarboxylic acid cycle. ZSS at dosage of 15 g/kg and 30 g/kg both can significant callback 11 potential biomarkers. Conclusion These metabolic changes indicated that ZSS facilitated recovery from insomnia by regulating the amino acid metabolism and energy metabolism, which provides the basis for elucidating therapeutic mechanisms underlying the efficiency of ZSS for insomnia treatment.