基于网络药理学和UPLC-QE-MS探讨和肝汤治疗功能性消化不良的作用机制  

Mechanisms Study of Hegan Decoction in Treatment of Functional Dyspepsia Based on Network Pharmacology and UPLC-QE-MS

作　　者：赵静怡 郑金粟[1] 曹锐[1] ZHAO Jingyi;ZHENG Jinsu;CAO Rui(Beijing Chaoyang Hospital,Capital Medicine University,Beijing 100020,China)

机构地区：[1]首都医科大学附属北京朝阳医院,北京100020

出　　处：《中国中医基础医学杂志》2023年第4期631-637,641,共8页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：北京市医管局培育计划(PZ2022003)-和肝汤颗粒治疗功能性消化不良(脾虚气滞型)的临床研究;首都医科大学科研培育基金(PYZ21085)-和肝汤对功能性消化不良十二指肠微炎症和肠道菌群作用的研究;首都医科大学附属北京朝阳医院金种子基金(CYJZ202118)-基于网络药理学的和肝汤治疗功能性消化不良十二指肠微炎症的机制研究。

摘　　要：目的 通过网络药理学分析和分子对接以及超高效液相色谱-四级杆静电场轨道阱质谱(ultra performance liquid chromatography q exactive mass spectrometry, UPLC-QE-MS)非靶向代谢组学法,探讨和肝汤治疗功能性消化不良(functional dyspepsia,FD)的潜在作用机制。方法 首先,通过TCMSP、TCMID数据库提取和肝汤相关的活性成分;通过DisGeNET、CTD和GeneCards数据库提取FD相关基因靶点;采用Cytoscape3.7.0建立处方-活性成分-靶点网络;由STRING构建蛋白相互作用(Protein-Protein interaction,PPI)网;通过R进行GO分析和KEGG通路富集分析,并采用MOL2和AutoDock进行分子对接;运用UPLC-QE-MS技术对和肝汤活性成分进行分析。结果 共筛选出76个和肝汤活性成分和75个FD潜在治疗靶标。处方-活性成分-靶点网络显示:山柰酚、槲皮素、金合花素、木犀草素、桉树醇、丁香酚等8个活性成分,以及p53、前列腺素E受体2、核因子红系2相关因子2、胱天蛋白酶3、钾电压门控通道亚家族H成员2这5个靶点基因在网络中存在最多关联,而肿瘤坏死因子(tumor necrosis factor,TNF)和白介素(interleukin,IL)-8等在PPI网络中MCC度最高。富集分析提示TNF和IL-17信号通路可能是关键的信号通路。UPLC-QE-MS结果显示,和肝汤颗粒内共识别出644个成分,其中10个成分与候选成分相匹配。分子对接显示山柰酚和刺芒柄花素与TNF、IL-8、IL-1β和IL-2具有较强的结合能力。结论 和肝汤含有山柰酚和刺芒柄花素等多种活性成分,其可能通过包括TNF信号通路、IL-17信号通路等多个通路和TNF、IL-8、IL-1β和IL-2等多个靶点,发挥免疫炎症调节作用,进而对FD产生治疗作用。Objective To explore the potential therapeutic mechanisms of HeGanTang(HGT)in treating functional dyspepsia(FD)based on network pharmacological analysis,molecular docking and ultra performance liquid chromatography q exactive mass spectrometry(UPLC-QE-MS).Methods Related compounds were obtained from TCMSP and TCMID databases.Target genes related to FD were acquired from DisGeNET,CTD and GeneCards.The prescription-ingredient-target interaction networks were established by Cytoscape3.7.0.The PPI network was constructed by STRING.GO analysis and KEGG pathway enrichment analysis were performed via R packages.The ingredients of HGT were analyzed by UPLC-QE-MS methods.Molecular docking analysis was conducted by the MOL2 and AutoDock.Results A total of 76 ingredients of HGT and 75 targets of FD were identified.The compound-target network analysis indicated that the 8 ingredients,including kaempferol,quercetin,acacetin,naringenin,luteolin,Khell,eucalyptol and eugenol were linked to more than 30 genes,and 5 genes(p53,PTGER2,NFE2L2,CASP3,KCNH2)were regulated by more than 30 active ingredients in the network.TNF and CXCL presented the highest MCC degree in the PPI network.Enrichment analysis indicated that TNF and IL-17 signaling pathways might be the critical signaling pathways.As the UPLC-QE-MS results showed,a total of 644 ingredients were recognized within HGT granules of which 10 ingredients were matched compared with the candidate ingredients.Molecular docking showed that kaempferol and formononetin had robust binding abilities with TNF,CXCL8,IL-1βand IL-2.Conclusion HGT contains kaempferol and formononetin,which may play a role in the regulation of immune inflammation through multiple pathways including TNF signaling pathway,IL-17 signaling pathway and multiple targets such as TNF,IL-8,IL-1βand IL-2,thus producing therapeutic effects on FD.

关 键 词：功能性消化不良 和肝汤 分子对接 网络药理学 炎性细胞因子 UPLC-QE-MS 

分 类 号：R285[医药卫生—中药学]