基于网络药理学探究宽胸气雾剂治疗心绞痛的作用机制

Network Pharmacology-Based Analysis on Pharmacological Mechanism of Kuanxiong Aerosol for Angina Pectoris Therapy

目的本研究运用网络药理学技术分析和筛选宽胸气雾剂治疗心绞痛的潜在作用靶点和效应机制,并通过药理实验初步阐明宽胸气雾剂治疗心绞痛的作用机制。方法采用BATMAN-TCM数据库建立宽胸气雾剂的"药物-靶点-通路-疾病"网络,TCMSP数据库筛选潜在活性分子及靶标信息,CTD数据库收集心绞痛相关基因,STRING平台构建蛋白质-蛋白质相互作用关系(PPI)网络,Cytoscape软件对PPI网络进行合并,通过MCODE插件筛选关键靶点,DAVID数据库进行基因本体(GO)、京都基因和基因组百科全书(KEGG)通路富集分析。根据预测结果进行药理实验,建立垂体后叶素(Pit)和异丙肾上腺素(ISO)诱导的心绞痛大鼠模型,通过心电图、血压、心率、心肌酶谱和病理切片研究宽胸气雾剂防治心绞痛的药理作用。结果网络药理学分析表明宽胸气雾剂治疗心绞痛的关键靶标包括一氧化氮合酶2(NOS2)、血红素加氧酶1(HO-1)、超氧化物歧化酶1(SOD1)、前列腺素内过氧化物合成酶2(PTGS2或COX-2)、IL-6、基质金属蛋白酶2(MMP2)、表皮生长因子受体(EGFR)、血管细胞黏附分子1(VCAM-1)和血管内皮生长因子A(VEGFA)等基因,主要富集在肿瘤坏死因子(TNF)、缺氧诱导因子(HIF-1)和磷脂酰肌醇3-激酶-蛋白激酶B(PI3K-Akt)等多条信号通路,涉及调控细胞凋亡、对机械刺激、缺氧、过氧化氢、脂多糖(LPS)的反应、一氧化氮(NO)合成和平滑肌细胞增殖等机制。药理实验证明宽胸气雾剂可减轻心肌缺血对心肌细胞造成的损伤,降低血清肌酸激酶(CK)、肌苷激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)活性,使病理状态减轻,但对于血压和心率无明显影响。结论宽胸气雾剂能够通过多个靶点和多条信号通路以非血液动力学特征防治心绞痛。

Objective Network pharmacology technology was applied to identify and screen the potential targets of Kuanxiong Aerosol and their functions,and underlying mechanism was verified by following pharmacological experiments.Methods Ingredient-target-pathway-disease association networks were established by Bioinformatics analysis tool for molecular mechanism of traditional Chinese medicine(BATMAN-TCM).The active ingredients and associated targets were obtained using traditional Chinese medicine systems pharmacology database and analysis platform(TCMSP).Protein-protein interactions(PPI) networks of targets of Kuanxiong Aerosol and angina pectoris collected from comparative toxicogenomics database(CTD) were captured through STRING database.Networks were constructed and merged by Cytoscape software,and potential targets were recognized by the molecular complex detection(MCODE) plugin.Gene Ontology(GO) and Kyoto Gene and Genomic Encyclopedia(KEGG) pathway enrichment analysis were performed by Database for Annotation,Visualization and Integrated Discovery(DAVID).Electrocardiographs(ECG),blood pressure,heart rate,serum myocardial enzymes,and pathological study were carried out on the basis of analysis results to assess the pharmacological effects of Kuanxiong Aerosol on myocardial ischemia rat models induced by pituitrin(Pit) and isoprenaline(ISO).Results Network pharmacology analysis showed that potential targets mainly involved the regulation of nitric oxide synthase 2(NOS2),heme oxygenase 1(HO-1),superoxide dismutase 1(SOD1),prostaglandin-endoperoxide synthase 2(PTGS2 or COX-2),interleukin-6(IL-6),matrix metallopeptidase2(MMP2),epidermal growth factor receptor(EGFR),vascular cell adhesion molecule 1(VCAM-1),and vascular endothelial growth factor a(VEGFA),which mediated the process of apoptotic,cellular response to mechanical stimulus,hypoxia,lipopolysaccharide(LPS),and hydrogen peroxide,biosynthetic process of nitric oxide(NO),and regulation of smooth muscle cell proliferation.Multiple pathways such as tumor necrosis factor(TNF),hypoxia-inducible factor 1(HIF-1),and phosphatidylinositol-3-kinase-protein kinase B(PI3 K-Akt) were closely related to the pharmacological mechanism.Experimental results showed that rats treated with Kuanxiong Aerosol exhibited lower creatine kinase(CK),creatine kinase-MB(CK-MB),and lactate dehydrogenase(LDH) activity in serum and mitigated pathological phenotypes with no obvious change on blood pressure and heart rate,indicating protective effects of Kuanxiong Aerosol on acute myocardial ischemia injury.Conclusion Kuanxiong Aerosol could prevent angina with non-hemodynamic features through multiple targets and multiple signaling pathways.