超高效液相色谱串联四极杆飞行时间质谱联合网络药理学及分子对接技术分析蜂胶总黄酮治疗牙周炎的作用机制  

UPLC-Q-TOF/MS combined with network pharmacology and molecular docking technology to analyze the mechanism of total flavonoids in propolis for treatment of periodontitis

作　　者：汤毛毛 李治 王雨宵 陈博文 郭健 常相伟 桂双英 TANG Maomao;LI Zhi;WANG Yuxiao;CHEN Bowen;GUO Jian;CHANG Xiangwei;GUI Shuangying(School of Pharmacy,Anhui University of Chinese Medicine,Institute of Pharmaceutics,Anhui Acade-my of Chinese Medicine,Engineering Technology Research Center of Modernized Pharmaceutics An-hui Education Department(AUCM),Anhui Province Key Laboratory of Pharmaceutical Technology and Application(AUCM),Hefei,Anhui 230012,China)

机构地区：[1]安徽中医药大学药学院,安徽省中医药科学院药物制剂研究所,现代药物制剂安徽省教育厅工程技术研究中心,药物制剂技术与应用安徽省重点实验室,安徽合肥230012

出　　处：《安徽医药》2023年第5期901-907,I0001,I0002,共9页Anhui Medical and Pharmaceutical Journal

基　　金：国家自然科学基金项目(81873019)。

摘　　要：目的 系统描述蜂胶总黄酮(TFP)中主要化学成分,探讨蜂胶总黄酮治疗牙周炎的作用机制。方法 采用超高效液相色谱串联四极杆飞行时间质谱(UPLC-Q-TOF/MS)明确TFP的化学成分。利用PubChem与Swiss target prediction数据库获取TFP作用靶点,通过在线人类孟德尔遗传数据库(OMIM)和GeneCards数据库筛选牙周炎相关靶点,并通过Venny 2.1平台获取TFP与牙周炎交集靶点。采用Cytoscape 3.7.2软件与STRING数据库分别构建“成分-靶点”网络图和交集靶点互作网络图,并利用DAVID数据库对交集靶点进行GO功能及KEGG通路富集分析。利用Autodock vina1.1.2软件对筛选的TFP潜在药效成分与核心靶点进行分子对接。体外建立LPS诱导的人牙周膜干细胞(hPDLSCs)炎症损伤模型,并验证TFP的抗炎活性。结果从TFP中共鉴定出13个黄酮类成分。共获得90个TFP与牙周炎交集靶点,其中RAC-α丝氨酸/苏氨酸蛋白激酶(AKT1)、前列腺素内过氧化物合酶(PTGS2)、表皮生长因子受体(EGFR)等为TFP治疗牙周炎的核心靶点。GO功能分析得到1 496个条目,涉及抗菌、抗炎以及调节过度免疫反应等;KEGG通路富集得到134个条目,涉及PI3K-AKT信号通路、雌激素信号通路及松弛素等关键信号通路,表明TFP可通过抗炎、促进成骨分化、抑制成脂分化并降低氧化应激水平来发挥牙周炎治疗作用。分子对接结果显示,TFP中5,7,4’-三羟基-3,6-二甲氧基黄酮、染料木素、木犀草素-5-甲醚、白杨黄素等成分与PTGS2、EGFR核心靶点的对接构象更稳定,亲和力更强,提示其是TFP治疗牙周炎的潜在药效成分。体外细胞实验进一步证实TFP能够抑LPS诱导的hPDLSCs细胞上清液中炎性因子[肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-17(IL-17)]水平,表明TFP具有良好的抗炎活性。结论 蜂胶总黄酮中多种成分通过抗炎、减缓氧化应激损伤以及促进骨生成等多途径发挥牙周�Objective To systematically characterize the main chemical components of total flavonoids in propolis(TFP),and to explore the mechanism of action of TFP in the treatment of periodontitis.Methods Ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometer(UPLC-Q-TOF/MS) was used to determine the chemical composition of TFP.Using Pubchem and Swiss Target Prediction databases to search the potential targets of TFP.In addition,periodontitis-related targets were screened through the online human Mendelian genetic database(OMIM) and GeneCards databases,and the intersection targets of TFP and periodontitis were obtained by Venny 2.1 platform.Cytoscape 3.7.2 software and STRING database were used to construct the "component-target" network diagram and intersection target interaction network diagram,GO function and KEGG pathway enrichment analysis of intersection targets were performed using the DAVID database.Autodock vina1.1.2 software was used to perform molecular docking of the screened potential pharmacodynamic components of TFP with the core target.LPS-induced inflammatory model of human periodontal ligament stem cells(hPDLSCs) was established in vitro,and the anti-inflammatory activity of TFP was verified.Results Thirteen components were identified from TFP.A total of 90 intersections of TFP and periodontitis were obtained.Among them,serine/threonine-protein kinase 1(AKT1),prostaglandin-endoperoxide synthase(PTGS2),Epidermal growth factor receptor(EGFR)were the core targets of TFP in the treatment of periodontitis.GO function analysis yielded 1 496 items,involving antibacterial,anti-inflammatory,and regulation of excessive immune response,etc.The KEGG pathway was enriched with 134 entries,involving key signaling pathways such as PI3K-AKT signaling pathway,estrogen signaling pathway,relaxin pathway,etc.It showed the treatment of periodontitis through anti-inflammatory,promoting osteogenic differentiation,inhibiting lipogenic differentiation and reducing the level of oxidative stre

关 键 词：蜂胶 总黄酮 牙周炎 网络药理学 分子对接 骨生成 

分 类 号：R285[医药卫生—中药学]