基于^(1)H NMR代谢组学技术结合网络药理学的恒山黄芪抗疲劳药效比较及调控牛磺酸代谢的机制初探  

Characterization of the anti-fatigue effect of Astragali Radix from the Shanxi Hengshan area and its association with taurine metabolism based on^(1)H NMR metabolomic technology combined with network pharmacology

作　　者：但春凤 张庆瑜 何盼[1,2,3] 秦雪梅 李爱平[1,2,3] DAN Chun-feng;ZHANG Qing-yu;HE Pan;QIN Xue-mei;LI Ai-ping(Modern Research Center for Traditional Chinese Medicine,Shanxi University,Taiyuan 030006,China;The Key Laboratory of Effective Substances Research and Utilization in TCM of Shanxi Province,Taiyuan 030006,China;The Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education,Shanxi University,Taiyuan 030006,China)

机构地区：[1]山西大学中医药现代研究中心,山西太原030006 [2]地产中药功效物质研究与利用山西省重点实验室,山西太原030006 [3]山西大学化学生物学与分子工程教育部重点实验室,山西太原030006

出　　处：《药学学报》2023年第7期1751-1760,共10页Acta Pharmaceutica Sinica

基　　金：山西省深度贫困县科技精准扶贫专项(2019FP-03);山西省卫生健康委员会“十大晋药”中药材质量标准规范制定和综合利用研究项目(Zyczl2020001)。

摘　　要：比较评价恒山黄芪的抗疲劳药效并初步揭示其通过调控牛磺酸代谢发挥抗疲劳的分子机制。本研究首先比较评价了广灵黄芪与浑源黄芪的药效差异,其次采用^(1)H NMR代谢组学技术并结合统计学的方法对不同组小鼠腓肠肌组织代谢产物进行分析与指认,找出受黄芪调控的差异代谢物,并借助Metscape预测其上游靶标。通过整合课题组前期基于网络药理学的黄芪抗疲劳潜在靶点,采用Cytoscape软件构建黄芪潜在抗疲劳靶点-共有靶点-差异代谢物相关靶点网络图,通过度值(degree)筛选关键靶点。最后依据共有靶点反推确定关键差异代谢物,并将其输入Metaboanalyst网站中进行通路富集分析,初步阐明分子机制。结果显示,黄芪干预能够明显改善小鼠力竭游泳时间,升高肝糖原并降低尿素氮水平,且广灵黄芪与浑源黄芪药效差异较小,即同属恒山山脉所产的黄芪质量较均一。进一步小鼠腓肠肌组织提取物指纹共指认34种代谢物,统计结果表明19个差异代谢物在恒山黄芪干预后发生显著回调。结合网络药理学最终明确恒山黄芪主要通过调控GAD1干预牛磺酸及次牛磺酸代谢来发挥抗疲劳作用。本研究结合^(1)H NMR代谢组学技术与网络药理学比较评价了恒山黄芪的抗疲劳药效,并初步揭示了其调控牛磺酸代谢抗疲劳的分子机制,为恒山黄芪的资源利用以及抗疲劳相关产品研发提供理论依据。本文涉及的动物实验操作均遵循山西大学动物伦理委员会的规定并通过动物实验伦理审查(批准号:SXULL2021028)。The objective of this work was to evaluate the anti-fatigue efficacy of Astragali Radix(AR)from the Shanxi Hengshan area and to reveal possible mechanisms by which it relieves fatigue.Efficacy differences between Guangling(GL)and Hunyuan(HY)AR preparations were compared and evaluated,and an^(1)H NMR metabolomic technique combined with statistical methods was used to identify the metabolites in different groups of mouse gastrocnemius muscle tissues.The differential metabolites after AR treatments were identified according to VIP and P values and the upstream targets were predicted with the help of Metscape.Cytoscape software was utilized to construct a network map of AR potential anti-fatigue targets.Key differential metabolites were identified based on shared targets and entered into the Metaboanalyst website for pathway enrichment analysis,which led to the preliminary elucidation of the molecular mechanisms.The results showed that intervention with AR can significantly improve the swimming-to-exhaustion time,increase liver glycogen,and reduce urea-nitrogen levels in mice.The difference between GL and HY ARs was relatively small,indicating that the quality of AR produced in the Hengshan area is consistent and stable.The metabolic fingerprints of mouse gastrocnemius muscle tissue extracts were composed of 34 metabolites,and the statistical results showed that 19 differential metabolites were significantly reversed after the Hengshan AR intervention.We found that the anti-fatigue effects of AR in the Shanxi Hengshan area were mainly associated with taurine and hypotaurine metabolism through regulation of GAD1,based on network pharmacological analysis.In conclusion,^(1)H NMR metabolomic techniques were combined with network pharmacology to compare and evaluate the quality of Hengshan ARs,and further associate the fatigue relieve with the regulation of taurine metabolism.This provides a theoretical basis for the resource utilization of Hengshan ARs and the development of anti-fatigue-related products.The animal exper

关 键 词：恒山黄芪 抗疲劳 腓肠肌 代谢物组学 牛磺酸代谢 

分 类 号：R917[医药卫生—药物分析学]