基于血清代谢组学和网络药理学研究酸枣仁提取物治疗失眠的效应物质及作用机制  

Effective substance and mechanism of Ziziphi Spinosae Semen extract in treatment of insomnia based on serum metabolomics and network pharmacology

作　　者：卞振华 张文明 唐静月 费倩倩 胡敏敏[1] 陈晓伟 苏联麟 费程浩 季德[2] 毛春芹[2] 童黄锦[2] 陆兔林[2] 袁晓航[1] BIAN Zhen-hua;ZHANG Wen-ming;TANG Jing-yue;FEI Qian-qian;HU Min-min;CHEN Xiao-wei;SU Lian-lin;FEI Cheng-hao;JI De;MAO Chun-qin;TONG Huang-jin;LU Tu-lin;YUAN Xiao-hang(Department of Pharmacy,Wuxi Traditional Chinese Medicine Hospital Affliated to Nanjing Unioersity of Chinese Medicine,Wuxi 214071,China;College of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学无锡附属医院药学部,江苏无锡214071 [2]南京中医药大学药学院,江苏南京210023

出　　处：《中国中药杂志》2022年第1期188-202,共15页China Journal of Chinese Materia Medica

基　　金：无锡市科技发展资金项目(N20192038);无锡市卫生健康委科研项目(MS201941);无锡市卫健委中青年拔尖人才计划项目(HB2020059);江苏省研究生科研创新计划项目(KYCX20_1513);国家重点研发计划项目(2018YFC1707000);国家自然科学基金项目(81873008)。

摘　　要：基于血清代谢组学和网络药理学研究酸枣仁提取物治疗失眠的效应物质及作用机制。建立对氯苯丙氨酸(p-chlorophenylalanine,PCPA)诱导大鼠失眠模型,酸枣仁提取物灌胃给药后,进行一般形态观察、戊巴比妥钠诱导失眠实验和病理组织学等药效学评价。采用UHPLC-MS代谢组学技术,结合多元数据统计筛选酸枣仁提取物治疗失眠的潜在生物标志物,进一步分析相关代谢通路。采用UHPLC-Q-Exactive-MS/MS技术结合网络药理学,构建酸枣仁治疗失眠的"成分-靶点-通路"网络,探讨酸枣仁治疗失眠的效应物质及作用机制。戊巴比妥钠诱导睡眠实验和组织病理学HE(hematoxylin and eosin)染色结果表明酸枣仁提取物具有较好的治疗失眠作用。血清代谢组学共筛选出21个酸枣仁提取物治疗失眠的内源性生物标志物,获取了苯丙氨酸、酪氨酸和色氨酸生物合成,苯丙氨酸代谢,烟酸和烟酰胺代谢等代谢通路。UHPLC-Q-Exactive-MS/MS鉴定了酸枣仁提取物中34个化学成分,包括24个黄酮类成分、2个三萜皂苷类成分、4个生物碱成分、2个三萜酸成分以及2个脂肪酸成分。网络药理学分析表明酸枣仁治疗失眠主要作用于DRD2(dopamine D2 receptor)、HTR1A(5-hydroxytryptamine receptor 1A)及ADRA2A(alpha-2A adrenergic receptor)等靶蛋白;与神经活性配体受体相互作用、5-羟色胺能突触、钙信号等通路密切相关;酸枣仁中木兰花碱、N-去甲荷叶碱、山矾碱、油酸、棕榈酸、乌药碱、白桦脂酸以及美洲茶酸为其治疗失眠的潜在效应物质。该研究揭示了酸枣仁提取物治疗失眠通过多个代谢途径,整体上纠正紊乱的代谢轮廓,阐释了酸枣仁治疗失眠的多成分、多靶点、多途径整合机制,为深入研究酸枣仁治疗失眠的作用机制奠定基础,也为开发治疗失眠的创新中药提供支撑。This study aims to study the effective substance and mechanism of Ziziphi Spinosae Semen extract in the treatment of insomnia based on serum metabolomics and network pharmacology.The rat insomnia model induced by p-chlorophenylalanine(PCPA)was established.After oral administration of Ziziphi Spinosae Semen extract,the general morphological observation,pentobarbital sodium-induced sleep test,and histopathological evaluation were carried out.The potential biomarkers of the extract in the treatment of insomnia were screened by ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS)combined with multivariate analysis,and the related metabolic pathways were further analyzed.The"component-target-pathway"network was constructed by ultra-high performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry(UHPLC-Q-Exactive-MS/MS)combined with network pharmacology to explore the effective substances and mechanism of Ziziphi Spinosae Semen in the treatment of insomnia.The results of pentobarbital sodium-induced sleep test and histopathological evaluation(hematoxylin and eosin staining)showed that Ziziphi Spinosae Semen extract had good theraputic effect on insomnia.A total of 21 endogenous biomarkers of Ziziphi Spinosae Semen extract in the treatment of insomnia were screened out by serum metabolomics,and the metabolic pathways of phenylalanine,tyrosine and tryptophan biosynthesis,phenylalanine metabolism,and nicotinate and nicotinamide metabolism were obtained.A total of 34 chemical constituents were identified by UHPLC-Q-Exactive-MS/MS,including 24 flavonoids,2 triterpenoid saponins,4 alkaloids,2 triterpenoid acids,and 2 fatty acids.The network pharmacological analysis showed that Ziziphi Spinosae Semen mainly acted on target proteins such as dopamine D2 receptor(DRD2),5-hydroxytryptamine receptor 1 A(HTR1 A),and alpha-2 A adrenergic receptor(ADRA2 A)in the treatment of insomnia.It was closely related to neuroactive ligand-receptor interaction,serotonergic synapse,and calcium signalin

关 键 词：酸枣仁 失眠 内源性代谢物 生物标志物 代谢通路 效应物质 代谢组学 网络药理学 

分 类 号：R285[医药卫生—中药学]