基于分子对接和网络药理学探讨参附注射液有效活性成分治疗心力衰竭的作用研究

The effect of Shenfu injection on heart failure based on molecular docking and network pharmacology

目的 利用网络药理学和分子对接技术探讨参附注射液活性成分对心力衰竭(heart failure, HF)的治疗作用,并分析其可能的分子机制,为今后实验研究提供一定的理论参考。方法 检索中药系统药理学数据库和分析平台(TCMSP)并筛选参附注射液有效活性成分潜在的作用靶点。利用GeneCards、OMIM、PharmGkb、TTD和DrugBank等数据库检索疾病相对应的靶点。将筛选所得中药靶点和疾病靶点取交集得到中药—疾病共同靶点及韦恩图。利用Cytoscape软件构建中药成分-疾病靶点调控网络图。运用STRING在线数据库构建中药—疾病共同靶点的蛋白互作网络关系图,使用CytoNCA作图对核心目标基因进行筛选。根据基因连接度选取靠前的核心基因,同时查找基因对应的药物分子,利用Autodock vina软件进行分子对接。最后将中药—疾病共同靶点进行GO富集功能分析和KEGG信号通路富集分析,富集得到有效的基因和信号通路。结果 从TCMSP数据库筛选获得参附注射液治疗HF的有效活性成分43个和中药—疾病共同作用靶点97个。关键靶点包括CASP3、JUN、AKT1、MAPK8、PTGS2、AHR、TNF、IL1B和RELA。GO功能富集分析显示参附注射液的分子功能主要与核受体的活动、配体激活、转录因子的活动、G蛋白偶联受体活性、药物结合、多肽结合、G蛋白偶联、神经递质受体活性、突触后神经递质受体活性、类固醇激素受体活性和酰胺结合等相关。KEGG显示主要作用途径包括脂质和动脉粥样硬化、血流剪切应力和动脉粥样硬化、TNF信号通路、糖尿病并发症中AGE-RAGE(晚期糖基化终产物及其核受体)信号通路和C型凝集素受体信号通路等。蛋白互作网络分析和分子对接结果显示,参附注射液有效活性成分山柰酚、β-谷甾醇和人参皂苷Rh2脂质体(Rh2-L)与CASP3、JUN、AKT1、MAPK8、PTGS2、AHR、TNF、IL1B和RELA具有良好的亲和力;由此推断这些基因和信号通路可能发挥关键性的作用。结论 参附注射液中的有效活性成分可通过多靶点、多基因、多信号通路影响HF的发生发展。为后续实验研究提供线索和导向。

Objective To investigate the therapeutic effects of Shenfu injection on heart failure(HF) by network pharmacology and molecular docking techniques, and to analyze its possible molecular mechanism, so as to provide theoretical reference for future experimental studies. Methods The pharmacologic database and analysis platform of TCM system(TCMSP) were searched and the potential targets of the active components and active components of SFI were screened.GeneCards, OMIM, PharmGkb, TTD and DrugBank databases were used to search for diseases and their corresponding targets.The intersection of the screened TCM target and the disease target was establish to obtain the common TCM-disease target and Venn diagram. Cytoscape software was used to construct the network diagram of TCM component-disease target regulation.The protein interaction network diagram of the common target of TCM and disease was constructed by using STRING online database, and the core target genes were screened by using CytoNCA.The top core genes were selected according to gene connectivity, and drug molecules corresponding to genes were searched.Autodock vina software was used for molecular docking. Finally, GO enrichment function analysis and KEGG signal pathway enrichment analysis were performed on the common target of TCM and disease to obtain effective genes and signal pathways. Results Forty-three active components and 97 targets of TCM and disease of SFI in treating heart failure were screened from TCMSP database. Key targets include CASP3,JUN,AKT1,MAPK8,PTGS2,AHR,TNF,IL1B and RELA. GO function enrichment analysis showed that the cords with the main molecular function of the injection and the activity of nuclear receptor and the ligand of the activation of transcription factors, activity, G protein coupled receptor activity, drug combination, polypeptide, G protein coupling, neurotransmitter receptor activity and postsynaptic activity of neurotransmitter receptors, steroid hormone receptor activity, amide bond, etc.KEGG showed that the main pathways include lipids and atherosclerosis, blood flow shear stress and atherosclerosis, TNF signaling pathway, age-rage(advanced glycation end products and their nuclear receptors)signaling pathway in diabetic complications, and C-type lectin receptor signaling pathway. Protein interaction network analysis and molecular docking result showed that kaempferol, β-sitosterol and ginsenoside Rh2 liposome(RH2-L) had good affinity with CASP3,JUN,AKT1,MAPK8,PTGS2,AHR,TNF,IL1 B and RELA.It follows that these genes and signaling pathways may play key roles. Conclusion The active components in SFI can affect the occurrence and development of heart failure through multi-target, multi-gene and multi-signal pathway and provide clues and guidance for the follow-up experimemtal research.