基于网络药理学及分子对接技术探析青风藤干预强直性脊柱炎的分子作用机制

Exploration of the Molecular Mechanism of Caulis Sinomenii Intervention in Ankylosing Spondylitis Based on Network Pharmacology and Molecular Docking Technology

目的:运用网络药理学方法构建青风藤的化学成分-疾病靶点-代谢信号通路网络,从而分析青风藤干预强直性脊柱炎(AS)的分子作用机制。方法:通过检索相关文献,并运用TCMSP数据库和SwissTargetPrediction数据库获取青风藤主要活性成分和作用靶点,利用DrugBank、GeneCards、TTD等数据库获取与AS相关的靶基因;整合药物与疾病的交集靶点,以String数据库对核心靶点构建蛋白互作网络(PPI)分析;最后通过DAVID数据库对核心靶点进行基因本体(GO)功能富集分析和京都基因与基因组百科全书(KEGG)信号通路分析,采用Cytoscape3.6.0构建青风藤活性成分-AS作用靶点-代谢信号通路的可视化网络。结果:通过筛选获得青风藤潜在活性成分共6个,得到可作用于AS的靶点37个,核心靶点涉及溶质载体家族6(神经递质转运蛋白),成员4(SLC6A4)、多巴胺受体D2(DRD2)、5-羟色胺受体3A(HTR3A)、前列腺素G/H合酶2(PTGS2)、细胞色素P450 2D6(CYP2D6)等。GO功能富集分析共得到150个条目(P<0.05),其中生物过程98个,细胞组成19个,分子功能条目33个。KEGG通路分析富集得到13条信号通路(P<0.05),主要包括神经活性配体-受体相互作用、钙信号通路、血清素激活突触通路、环磷酸腺苷(cAMP)信号通路、血管平滑肌收缩、环磷酸鸟苷-依赖性蛋白激酶(cGMP-PKG)信号通路等;分子对接研究证实青风藤中大多数活性成分与治疗AS的靶点有着较好的结合活性。结论:基于网络药理学的研究表明,青风藤中千金藤啶碱、青藤碱等成分是治疗AS的关键成分,这些成分可能调节PTGS2、SLC6A4等靶点,并作用于血清素激活突触、 Janus激酶/信号转导与转录激活因子(JAK-STAT)等多条信号通路发挥发挥抑制炎症反应、调节免疫功能等作用来治疗AS,实现多成分、多靶点、多通路发挥对AS的治疗作用,具有一定的临床应用价值。

Objective: To analyze the molecular mechanism of Caulis Sinomenis intervention in ankylosing spondylitis(AS) by using network pharmacology to construct the chemical composition-disease target-metabolic signaling pathway network of Caulis Sinomenii. Methods: Through searching the related literature, TCMSP database and Swiss TargetPrediction database were used to obtain the main active ingredients and targets of Caulis sylvestris, and DrugBank, GeneCards, TTD and other databases were used to obtain the target genes related to AS. The intersection of drug and disease targets was integrated, and String database was used to construct protein interaction network(PPI) analysis of core targets. Finally, the core targets were analyzed by gene ontology(GO) function enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) signal pathway analysis through the DAVID database, and Cytoscape3.6.0 was used to construct the active ingredients of Caulis sylvestris-AS action targets-a visual network of metabolic signaling pathways. Results: A total of 6 potential active components of Caulis Sinomenii, and 37 targets that could act on AS were obtained. The core targets involved SLC6 A4, DRD2, HTR3 A, PTGS2 and CYP2 D6, etc. The GO function enrichment analysis yielded a total of 150 entries(P<0.05), including 98 biological processes, 19 cell compositions, and 33 molecular function entries. KEGG pathway analysis enriched 13 signaling pathways(P<0.05), mainly including neuroactive ligand-receptor interaction, calcium signaling pathway, serotonergic synapse, cAMP signaling pathway, vascular smooth muscle contraction and cGMP-PKG signaling Access etc. Molecular docking studies confirmed that most of the active ingredients in Caulis sinomenii had good binding activity with the targets for the treatment of AS. Conclusion: The studies based on network pharmacology show that stepholidine and sinomenine are the key components in the treatment of AS. These components may regulate targets such as PTGS2 and SLC6 A4, and act on serotonin-activated synapses, JAK-STAT and other signal pathways to inhibit inflammation and regulate immune function to treat AS, and to achieve the therapeutic effects of multi-component, multi-target and multi-pathway on AS. It has certain clinical application value.