LC-MS结合网络药理学探讨“片姜黄-当归”挥发油防治膝骨关节炎的作用机制

LC-MS Combined with Network Pharmacology to Explore the Mechanism of “Wenyujin Rhizoma Concisum-Angelicae Sinensis Radix” Essential Oil in the Treatment of Knee Osteoarthritis

关键词:  挥发油  片姜黄  当归  化学成分  膝骨关节炎  液相色谱-质谱法  网络药理学

Key words:  essential oil  Wenyujin Rhizoma Concisum  Angelicae Sinensis Radix  chemical composition  knee osteoarthritis  liquid chromatograph mass spectrometer  network pharmacology

摘要:

目的 提取“片姜黄-当归”药对挥发油，使用液相色谱-质谱联用仪(LC-MS)对提取的挥发油进行化学成分定性分析，并运用网络药理学探讨药对挥发油防治膝骨关节炎(knee osteoarthritis，KOA)的作用机制。方法 采用水蒸气蒸馏法提取“片姜黄-当归”药对挥发油，并使用LC-MS对挥发油成分进行分析。借助Pubchem、Swiss Target Prediction、GeneCards、DAVID等数据库预测有效成分防治KOA的作用靶点及通路，对核心基因进行基因本体(Gene ontology，GO)分析与京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes，KEGG)通路富集分析，并使用Cytoscape软件构建“活性成分-疾病靶点”可视化网络图。运用AutoDock及Pymol对度值靠前的靶点与活性成分进行分子对接验证。结果 “片姜黄-当归”药对挥发油中共鉴定出59种化合物，主要成分为莪术酮、异姜黄醇、藁本内酯等；通过网络药理学筛选出挥发油作用于KOA的28个潜在靶点，包括CTSK、PTGS1、PTGS2和ESR1等关键靶点；KEGG富集分析预测“片姜黄-当归”药对挥发油主要通过肿瘤坏死因子信号通路、破骨细胞分化、白介素17信号通路、松弛素信号通路、丝裂原活化蛋白激酶信号通路来发挥治疗KOA的作用。分子对接结果显示筛选的活性成分与其对应靶蛋白均具有较好的结合活性。结论 使用LC-MS结合网络药理学的方法初步探明了“片姜黄-当归”药对挥发油中的主要有效成分及其干预KOA的潜在靶点与作用机制，为“片姜黄-当归”后续的深入研究提供思路。

Abstract:

OBJECTIVE To extract the essential oil of “Wenyujin Rhizoma Concisum-Angelica Sinensis Radix”, and to qualitatively analyze the chemical composition of the extracted essential oil by LC-MS. Network pharmacology was used to explore the mechanism of the essential oil acting on knee osteoarthritis(KOA). METHODS The essential oil of “Wenyujin Rhizoma Concisum-Angelicae Sinensis Radix” was extracted by steam distillation, the components of the essential oil were analyzed by LC-MS. PubChem, Swiss Target Prediction, GeneCards, DAVID and other databases were used to predict disease targets and pathways of active component in prevention and treatment of KOA. The key genes were analyzed by Gene ontology(GO) analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis. Cytoscape was used to construct a visual network map of “active component-disease targets”. AutoDock and Pymol were used to verify the molecular docking between the target with the highest degree target and the active components. RESULTS A total of 59 compounds were identified in the “Wenyujin Rhizoma Concisum-Angelicae Sinensis Radix” essential oil, and the main components were curzerenone, isocurcumenol, ligustilide, etc. Through network pharmacology, 28 potential targets of essential oil acting on KOA were screened, including CTSK, PTGS1, PTGS2 and ESR1, etc. KEGG enrichment analysis predicted that “Wenyujin Rhizoma Concisum-Angelicae Sinensis Radix” essential oil mainly targeted TNF signaling pathway, osteoclast differentiation, IL-17 signaling pathway, relaxin signaling pathway, MAPK signaling pathway to treat KOA. The results of molecular docking showed that the screened active components and their corresponding target proteins had positive binding activity. CONCLUSION LC-MS combined with network pharmacology is used to preliminaries explore the main active components and the potential targets and mechanism of action of “Wenyujin Rhizoma Concisum-Angelicae Sinensis Radix” essential oil in the intervention of KOA, providing ideas for the further study of “Wenyujin Rhizoma Concisum-Angelicae Sinensis Radix” essential oil.