基于网络药理学和分子对接技术探讨石菖蒲治疗缺血性脑卒中的作用机制

Mechanism of action of Rhizoma Acori graminei in treatment of cerebral ischemic stroke based on network pharmacology and molecular docking

目的:借助网络药理学和分子对接技术研究石菖蒲治疗缺血性脑卒中(CIS)的作用机制。方法:以"石菖蒲"为关键词在中药系统药理学数据库与分析平台(TCMSP)检索其活性成分和作用靶点,利用Uniprot数据库对靶点蛋白信息进行标准化,构建中药、成分、靶点网络。利用GeneCards、TTD、OMIM和DRUGBANK 4个数据库检索CIS相关疾病靶点。将疾病和药物靶点导入微生信软件绘制韦恩图得到交集靶点,使用在线软件String 11.0建立蛋白质相互作用(PPI)网络。采用Cytoscape 3.7.2软件对PPI网络进行可视化分析,筛选出核心靶点。在Metascape数据库对核心靶点进行GO富集分析及KEGG通路分析。最后采用分子对接技术确定石菖蒲的有效成分作用于CIS的核心靶点。结果:获得38个潜在作用靶点,PPI网络拓扑分析发现RAC-α丝氨酸/蛋白激酶(AKT1)、肿瘤坏死因子(TNF)可能是石菖蒲治疗CIS的核心基因。GO富集分析结果显示石菖蒲可能是通过对血压的负向调节、调控白细胞迁移等炎症反应、神经递质合成过程、血管生成以及血液循环等生物学过程来发挥作用。KEGG通路富集分析表明,石菖蒲作用于CIS中的通路涉及血流剪切应力与动脉粥样硬化、钙信号通路、cAMP信号通路、VEG通路以及炎症和癌症等通路。分子对接技术验证了石菖蒲有效成分中的山柰酚与潜在靶点AKT1和TNF具有较好的结合活性。结论:石菖蒲的主要活性成分山柰酚可能通过作用于AKT1等多个靶点,调控钙信号通路等多条信号通路,从而发挥血管生成、抗神经细胞凋亡、抗炎、抑制细胞凋亡等作用,来达到预防或治疗CIS的目的。

Objective:To investigate the mechanism of action of Rhizoma Acori graminei in the treatment of cerebral ischemic stroke(CIS)based on network pharmacology and molecular docking.Methods:With“Rhizoma Acori graminei”as the keyword,Traditional Chinese Medicine Systems Pharmacology was searched to obtain its active components and action targets,and Uniprot database was used to standardize the information of target proteins,so as to construct a traditional Chinese medicine(TCM)drug-component-target network.GeneCards,TTD,OMIM,and DRUGBANK databases were used to search for the disease targets of CIS.The disease targets and the drug targets were imported into bioinformatics software to plot the Venn diagram and obtain intersecting targets,and the online software String 11.0 was used to construct a protein-protein interaction(PPI)network.Cytoscape 3.7.2 was used for visual analysis of the PPI network,and core targets were screened out.Gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were performed for the core targets in Metascape database.The molecular docking technique was used to identify the effective constituents of Rhizoma Acori graminei acting on the core targets of CIS.Results:A total of 38 potential action targets were obtained,and PPI network topology analysis showed that RAC-α serine/protein kinase(AKT1)and tumor necrosis factor(TNF)might be the core genes in the treatment of CIS by Rhizoma Acori graminei.GO enrichment analysis showed that Rhizoma Acori graminei exerted a therapeutic effect possibly by negatively regulating blood pressure and regulating various biological processes such as inflammatory response(including leukocyte traffic),neurotransmitter synthesis,angiogenesis,and blood circulation.KEGG pathway enrichment analysis showed that Rhizoma Acori graminei acted on CIS via the pathways including blood flow shear stress and atherosclerosis,calcium signaling pathway,cAMP signaling pathway,VEG signaling pathway,and inflammation/cancer signaling pathways.Molecular docking showed that kaempferol,one of the effective constituents of Rhizoma Acori graminei,had good binding activity to the potential targets AKT1 and TNF.Conclusion:As the main effective constituent of Rhizoma Acori graminei,kaempferol may act on various targets including AKT1,regulate various signaling pathways including the calcium signaling pathway,and thus achieve the prevention or treatment of CIS by promoting angiogenesis and inhibiting nerve cell apoptosis,inflammation,and cell apoptosis.