基于网络药理学与体外实验探讨杏仁活性成分治疗血管痉挛的作用机制

Mechanism of action of active components of Prunus armeniaca L.var.ansu Maxim.mature seed for vasospasm based on network pharmacology and in vitro experiments

目的运用网络药理学和体外细胞实验探究杏仁活性成分治疗血管痉挛的潜在作用机制。方法检索TCMSP数据库、Genecards数据库、SwissTargetPrediction数据库、Drugbank数据库等,获得杏仁的活性成分、作用靶点及血管痉挛相应疾病靶点,并将其作用靶点与获得的疾病靶点的交集作为潜在靶点;利用String平台,对潜在靶点构建蛋白-蛋白互作(PPI)网络;采用R语言软件包对潜在靶点进行基因本体(GO)功能及京都基因与基因组百科全书(KEGG)富集分析,然后使用Cytoscape软件分析其网络拓扑结构,筛选其核心靶点,并进行分子对接验证。在此基础上,通过MTT法检测苦杏仁苷对血管紧张素Ⅱ所致血管平滑肌细胞增殖的影响。结果杏仁治疗血管痉挛的主要潜在活性成分有17个;药物-疾病核心靶点有68个;涉及的信号通路主要有钙信号通路、神经活性配体-受体相互作用、鞘脂信号通路、血管平滑肌收缩、EGFR酪氨酸激酶抑制剂耐药性、松弛素信号通路等;关键核心靶点与活性成分苦杏仁苷、雌酚酮结合具有稳定构象。体外实验结果表明,苦杏仁苷能抑制血管紧张素Ⅱ所致血管平滑肌细胞增殖,其机制可能与其抑制MAPKs信号转导通路的活化有关。结论杏仁活性成分可能通过作用于基质金属肽酶9、雌酚酮等靶点治疗血管痉挛,为杏仁活性成分治疗血管痉挛的作用机制研究提供了参考。

Objective To explore the potential mechanism of the active components of Prunus armeniaca L.var.ansu Maxim.mature seed for vasospasm by integrating network pharmacology and in vitro cell experiments.Methods We searched TCMSP,Genecards,SwissTargetPrediction database,Drugbank database,and other related literatures to obtain active components in Prunus armeniaca L.var.ansu Maxim.mature seed,action targets Rhizoma,and corresponding disease targets.The intersection of action and disease targets was taken as the potential targets.A protein-protein interaction network(PPI)for potential targets was established based on String platform.R language software package was used to analyze the function of Gene Ontology(GO)and the enrichment of Kyoto Encyclopedia of Genes and Genomes(KEGG),and then Cytoscape software was used to analyze its network topology and further screen core targets.Molecular docking was used to verify the predicted results.Subsequently,the effect of active components in Prunus armeniaca L.var.ansu Maxim.mature seed on the proliferation of vascular smooth muscle cells(VSMCs)induced by angiotensinⅡ(AngⅡ)was detected by MTT.Results Totally 17 main potential active components of Prunus armeniaca L.var.ansu Maxim.mature seed in the treatment of vasospasm and 68 drug-disease targets were found.The main signaling pathways include calcium signaling pathway,neuroactive ligand-receptor interaction,sphingolipid signaling pathway,vascular smooth muscle contraction,EGFR tyrosine kinase inhibitor resistance,relaxin signaling pathway,etc.The stable conformations were formed between the core targets and the active components,amygdalin and estrone.The in vitro results showed that amygdalin inhibited AngⅡ-induced proliferation and phenotype transformation of VSMCs through inactivating the MAPKs signaling pathways.Conclusion Various active components in Prunus armeniaca L.var.ansu Maxim.mature seed may play a role in vasospasm by acting on targets such as matrix metallopeptidase 9(MMP9),estrone,etc.The study helps reveal the mechanism of active components of Prunus armeniaca L.var.ansu Maxim.mature seed in the treatment of vasospasm.