摘要
目的:分析川芎酒制前后水提物中差异性成分,探讨川芎酒制增强抗脑缺血损伤的分子作用机制。方法:采用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱法(UHPLC-Q-Orbitrap HRMS),基于化合物质谱信息并结合对照品、参考文献比对,定性分析川芎水提物的主要化学成分,采用化学模式识别方法筛选川芎酒制前后的差异性成分。基于此,通过在线数据库对差异化合物的潜在靶点进行预测,搜索脑缺血疾病相关靶点。运用Cytoscape 3.6.0建立川芎酒制前后差异成分-作用靶点-疾病网络图;使用STRING 11.5构建交集靶点蛋白质-蛋白质相互作用(PPI)网络;通过DAVID 6.8对差异性成分抗脑缺血的潜在靶点进行基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析;同时,将相对含量较高且酒制后峰面积升高的化合物与其对应靶点进行分子对接,以验证酒制增效机制。结果:从川芎中共鉴定出71个化学成分,筛选出34个差异性成分,对应潜在靶点603个,与769个疾病靶点做交集后共得到60个交集靶点。通过PPI网络分析得到7个关键靶点,包括JUN、信号转导和转录激活因子3(STAT3)、丝裂原活化蛋白激酶3(MAPK3)、白细胞介素-1β(IL-1β)、血管内皮生长因子A(VEGFA)、胱天蛋白酶-3(CASP3)、基质金属蛋白酶9(MMP9);肿瘤坏死因子(TNF)信号通路为主要的差异信号通路。分子对接结果显示,各差异成分(洋川芎内酯K、洋川芎内酯F、3-正丁基苯酞、Z,Z′-6,8′,7,3′-双藁本内酯、阿魏酸、Z-藁本内酯)和其对应靶点均具有较好的结合活性。结论:川芎酒制增效机制可能与抑制炎症反应的作用增强有关。
Objective:To analyze the differential components in water extract of Chuanxiong Rhizoma before and after processing with wine,and to explore the molecular mechanism of Chuanxiong Rhizoma processed with wine in enhancing anti-cerebral ischemia injury.Method:Ultra high performance liquid chromatography tandem quadrupole orbitrap high resolution mass spectrometry(UHPLC-Q-Orbitrap HRMS)was used to qualitatively analyze the main chemical components in water extract of Chuanxiong Rhizoma based on the spectral information of compound,comparison of reference substance and references.The chemical pattern recognition method was used to screen the differential components of Chuanxiong Rhizoma before and after processing.Based on these differential components,the potential targets of differential components were predicted by online databases,and the related targets of cerebral ischemia were searched.Cytoscape 3.6.0 was used to establish the network diagram of differential components-action targets-diseases of Chuanxiong Rhizoma processed with wine.The protein-protein interaction(PPI)network of intersection targets was constructed by STRING 11.5.The potential targets of differential components against cerebral ischemia were analyzed by Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis through DAVID 6.8.At the same time,the chemical compounds with high relative content and increased peak area after wine processing were docked with their corresponding targets to verify the mechanism of enhanced effect after wine processing.Result:A total of 71 chemical components were identified from Chuanxiong Rhizoma,34differential components and 603 potential targets were screened out.At the same time,a total of 769 disease targets and 60 intersection targets were obtained.Seven key targets were identified through PPI network analysis,including JUN,signal transducer and activator of transcription 3(STAT3),mitogen-activated protein kinase 3(MAPK3),interleukin-1β(IL-1β),vascular endothelial growth factor A(VEGFA),Caspase-3(CASP3)and mtrix metalloproteinase 9(MMP9).Tumor necrosis factor(TNF)signaling pathway was the main differential signaling pathway.The results of molecular docking showed that differential components(senkyunolide K,senkyunolide F,3-n-butylphthalide,Z,Z′-6,8′,7,3′-diligustilide,ferulic acid and Z-ligustilide)and corresponding targets had good binding activities.Conclusion:The synergistic mechanism of Chuanxiong Rhizoma processed with wine may be related to the enhanced inhibitory effect of inflammatory reaction.
作者
裴科
宁燕
蔡皓
曹丽珑
赵婷婷
于子涵
曹岗
王颖莉
张朔生
PEI Ke;NING Yan;CAI Hao;CAO Li-long;ZHAO Ting-ting;YU Zi-han;CAO Gang;WANG Ying-li;ZHANG Shuo-sheng(Shanxi Modern Chinese Medicine Engineering Laboratory,College of Traditional Chinese Medicine(TCM)and Food Engineering,Shanxi University of Chinese Medicine,Jinzhong 030619,China;Research Center of TCM Processing,College of Pharmacy,Zhejiang Chinese Medical University,Hangzhou 310053,China;Engineering Research Center for Standardization and Normalization of TCM Processing,Ministry of Education,School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing 210023,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第12期164-173,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81703704)
山西省重点研发计划项目(201903D421018)
中医脑病学山西省重点实验室课题(202105D121012)
浙江省重点建设高校优势特色学科(中药学)开放基金项目(ZYAOX2018023)
山西省应用基础研究项目(201801D221436)
山西省脑病重点实验室开放课题(CME-OP-201003)
山西省教育厅科技创新计划项目(2019L0720)
山西省中医药管理局项目(2019ZYYC016)
山西药茶的研制和开发项目(2020PY-YC-20)。
