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
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摘要
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.
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.
引文
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[22]李爱平,范玛莉,张松,等.山西老陈醋对大鼠机体影响的代谢组学研究[J].食品科学,2015,36(23):268-274.
[23]张敏,杜晨晖,马敏,等.酸枣仁饮片汤剂质量评价研究[J].中草药,2018,49(19):4520-4527.
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[27]张妍,解军波,张彦青,等.镇静催眠药物的主要作用机制研究进展[J].中国药房,2011,22(41):3906-3908.
[28]邓元江,刘卫英,梁伟雄,等.穴位埋线对青霉素致痫大鼠海马氨基酸类神经递质的影响[J].中华中医药学刊,2006,24(5):827-829.
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[30]Lee R S,Steffensen S C,Henriksen S J.Discharge profiles of ventral tegmental area GABA neurons during movement,anesthesia,and the sleep-wake cycle[J].JNeurosci,2001,21(5):1757-1766.
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[32]刁远明.安寐汤促睡眠药效研究及机理初探[D].广州:广州中医药大学,2010.
[33]许海玉,王松松,杨洪军,等.基于网络药理学探析复方阿胶浆辅助治疗肿瘤的作用机制研究[J].中国中药杂志,2014,39(16):3148-3151.
[34]Wishart D S,Craig K,Guo A C,et al.Drug Bank:Aknowledgebase for drugs,drug actions and drug targets[J].Nucl Acid Res,2008,doi:10.1093/nar/gkm958.
[35]Python A,Steimer T,Hilaire Z D S,et al.Extracellular serotonin variations during vigilance states in the preoptic area of rats:A microdialysis study[J].Brain Res,2001,doi:10.1016/s0006-8993(01)02477-5.
[2]Sarris J,Panossian A,Schweitzer I,et al.Herbal medicine for depression,anxiety and insomnia:A review of psychopharmacology and clinical evidence[J].Eur Neuropsychopharmacol,2011,21(12):841-860.
[3]Richey S M,Krystal A D.Pharmacological advances in the treatment of insomnia[J].Current Pharm Design,2011,17(15):1471-1475.
[4]郭岳.交泰汤治疗心肾不交型不寐临床观察[J].中草药,2013,44(9):1174-1175.
[5]张如意,游秋云,王平,等.酸枣仁汤对慢性睡眠剥夺老年失眠大鼠心肌细胞凋亡及相关蛋白Bcl-2、Bax表达的影响[J].中华中医药杂志,2017,32(4):1691-1693.
[6]张飞燕,李晶晶,周莹,等.安神类中药及其有效成分对神经递质镇静催眠机制的研究进展[J].中国中药杂志,2016,41(23):4320-4327.
[7]中国药典[S].一部.2015.
[8]曹文聪,海英,任路.基于数据挖掘中药复方治疗失眠用药规律分析[J].辽宁中医杂志,2018,45(9):1851-1853.
[9]吕辰鹏,王丽斐,何泉泉,等.改善睡眠功能保健食品的研究进展[J].食品工业,2017,38(8):238-243.
[10]Cao J X,Zhang Q Y,Cui S Y,et al.Hypnotic effect of jujubosides from Semen Ziziphi Spinosae[J].JEthnopharmacol,2010,130(1):163-166.
[11]Jiang J G,Huang X J,Chen J,et al.Comparison of the sedative and hypnotic effects of flavonoids,saponins,and polysaccharides extracted from Semen Ziziphus jujube[J].Nat Prod Res,2007,21(4):310-320.
[12]Ma Y,Han H,Nam S Y,et al.Cyclopeptide alkaloid fraction from Zizyphi Spinosi Semen enhances pentobarbital-induced sleeping behaviors[J].JEthnopharmacol,2008,117(2):318-324.
[13]Shergis J L,Ni X,Sarris J,et al.Ziziphus spinosa seeds for insomnia:A review of chemistry and psychopharmacology[J].Phytomed Int J Phytother Phytopharmacol,2017,doi:10.1016/j.phymed.2017.07.004.
[14]吴传芳.酸枣仁对地西泮的增效减毒作用研究[D].哈尔滨:黑龙江中医药大学,2017.
[15]翟旭峰,肖小春,娄勇军,等.生酸枣仁及其炮制品镇静催眠作用及对失眠大鼠脑电图的影响[J].中药药理与临床,2015,31(6):94-97.
[16]张舜波,王平,田代志,等.酸枣仁总皂苷对失眠老年大鼠脑氨基酸类神经递质及受体表达的影响[J].中国实验方剂学杂志,2014,20(4):124-127.
[17]张林挺.酸枣仁汤对虚劳失眠大鼠干预机理的研究[D].广州:广州中医药大学,2009.
[18]季文博.酸枣仁-五味子“药对”提取物镇静催眠作用的药效学及机制研究[D].合肥:安徽中医药大学,2013.
[19]张军武,田凡,邹文信,等.酸枣仁汤对DL-4-氯苯基丙氨酸所致失眠的药效学研究[J].河南中医,2016,36(6):985-987.
[20]张亚宁.基于步态分析的酸枣仁汤抗失眠、抗焦虑作用研究[D].咸阳:陕西中医学院,2014.
[21]陈金锋,高家荣,季文博,等.酸枣仁-五味子药对镇静催眠作用及机制研究[J].中药药理与临床,2013,29(4):128-131.
[22]李爱平,范玛莉,张松,等.山西老陈醋对大鼠机体影响的代谢组学研究[J].食品科学,2015,36(23):268-274.
[23]张敏,杜晨晖,马敏,等.酸枣仁饮片汤剂质量评价研究[J].中草药,2018,49(19):4520-4527.
[24]赵芳,李强,梁梅丽,等.超高效液相色谱-串联质谱法快速测定大鼠血清中8种神经递质[J].分析化学,2018,46(1):121-128.
[25]He W,Liu G,Cai H,et al.Integrated pharmacokinetics of five protoberberine-type alkaloids in normal and insomnic rats after single and multiple oral administration of JiaoTai-Wan[J].J Ethnopharmacol,2014,154(3):635-644.
[26]郑诏谦.原络配穴为主针药结合治疗心肾不交型失眠症的临床研究[D].广州:广州中医药大学,2012.
[27]张妍,解军波,张彦青,等.镇静催眠药物的主要作用机制研究进展[J].中国药房,2011,22(41):3906-3908.
[28]邓元江,刘卫英,梁伟雄,等.穴位埋线对青霉素致痫大鼠海马氨基酸类神经递质的影响[J].中华中医药学刊,2006,24(5):827-829.
[29]孙春宇.天王补心丸与磁朱丸对失眠大鼠睡眠时相影响的比较研究[D].哈尔滨:黑龙江中医药大学,2007.
[30]Lee R S,Steffensen S C,Henriksen S J.Discharge profiles of ventral tegmental area GABA neurons during movement,anesthesia,and the sleep-wake cycle[J].JNeurosci,2001,21(5):1757-1766.
[31]Manfridi A,Brambilla D,Mancia M.Sleep is differently modulated by basal forebrain GABA(A)and GABA(B)receptors[J].Am J Physiol Regul Integr Comp Physiol,2001,281(1):R170-R175.
[32]刁远明.安寐汤促睡眠药效研究及机理初探[D].广州:广州中医药大学,2010.
[33]许海玉,王松松,杨洪军,等.基于网络药理学探析复方阿胶浆辅助治疗肿瘤的作用机制研究[J].中国中药杂志,2014,39(16):3148-3151.
[34]Wishart D S,Craig K,Guo A C,et al.Drug Bank:Aknowledgebase for drugs,drug actions and drug targets[J].Nucl Acid Res,2008,doi:10.1093/nar/gkm958.
[35]Python A,Steimer T,Hilaire Z D S,et al.Extracellular serotonin variations during vigilance states in the preoptic area of rats:A microdialysis study[J].Brain Res,2001,doi:10.1016/s0006-8993(01)02477-5.