抗幽门螺杆菌活性挥发油的筛选、药效物质分析及其作用机制预测

Screening of anti-Helicobacter pylori active volatile oils,analysis of pharmacodynamic substances and prediction of their mechanism of action

目的探讨抗幽门螺杆菌(HP)活性挥发油的药效物质,初步预测潜在活性成分及其作用机制。方法微量稀释法(96微孔板)测定丁香、广藿香、紫苏、牛至、枳实、厚朴、薄荷、肉桂8种挥发油对HP的抑制率,筛选出抑菌活性最强的3种挥发油。气相色谱-质谱法联用(GC-MS)分析3种挥发油的化学成分,利用TCMSP、Swiss TargetPrediction、PubChem数据库获取3种挥发油主要活性成分及其对应靶点信息;网络药理学技术预测其潜在活性成分、相关通路及关键靶点;AUTODOCKVina软件对核心成分及关键靶点基因进行分子对接,初步验证潜在活性成分及关键靶点。结果广藿香、牛至和肉桂挥发油抑制90%细菌生长的最低药物浓度(MIC_(90))分别为0.250、0.250、0.125μL·mL^(-1),具有较强的抑菌活性;从以上3种挥发油中分别鉴定出27、26、15个化学成分,共筛选出15个活性成分,涉及232个基因靶点,影响了与抑制HP相关的癌症通路、化学致癌-受体激活及EGFR酪氨酸激酶抑制剂耐药性等275条信号通路;分子对接验证结果显示3种挥发油抗HP与SRC蛋白密切相关,15个活性成分与关键靶点SRC的结合较为稳定。结论广藿香、牛至、肉桂3种活性挥发油通过多成分、多靶点、多通路的形式发挥抗HP作用。

Objective To screen the volatile oils with anti-Helicobacter pylori(HP)activity from eight plant volatile oils,to explore their pharmacodynamic substances,and to preliminarily predict the potential active components and their mechanisms of action.Methods The inhibition rates of eight volatile oils against HP were determined by microdilution(96 microtiter plate),and the top three volatile oils with antibacterial activity were screened out.The chemical compositions of the three volatile oils were analyzed by Gas Chromatography-Mass Spectrometry(GC-MS),and the information of the three volatile oils and their corresponding targets were obtained by using TCMSP,Swiss Target Prediction,and Pub Chem databases;Network pharmacology techniques were used to predict the potential active ingredients,related pathways and key targets;AUTO DOCK vina software was used to perform molecular docking on the genes of the core ingredients and key targets,and the potential active ingredients and key targets were preliminarily verified.Results The minimum inhibitory concentration(MIC_(90))of volatile oils of Pogostemonis Herba,Origani Herba,and Cinnamomi Cortex were 0.250,0.250 and 0.125μL·mL^(-1),respectively,with strong bacteriostatic activity.The three volatile oils identified 27,26 and 15 chemical components respectively,and a total of 15 active components were screened,including 232 gene targets,affecting pathways in cancer,chemical carcinogenesis-receptor activation and EGFR tyrosine kinase inhibitor resistance(EGFR),which are related to the inhibition of HP.Comparative molecular docking validation showed that the three volatile oils against HP were closely related to SRC proteins,and the 15 active ingredients obtained from the screening showed more stable binding to the key target SRC.Conclusion The three active volatile oils(Pogostemonis Herba,Origani Herba,and Cinnamomi Cortex)exerted anti-HP effects through multi-components,multi-targets and multi-pathways,which provided a scientific basis for further elucidation of the anti-HP mechanism of volatile oils.