参附注射液调控应激反应的作用机制:基于网络药理学方法和分子对接技术

Mechanism of Shenfu injection in regulating stress response based on network pharmacology and molecular docking

目的通过网络药理学和分子对接技术探讨参附注射液经神经-内分泌-免疫系统调控应激反应的潜在作用机制。方法使用中药系统药理学数据库和分析平台筛选参附注射液的主要活性成分及作用靶点,使用PharmMapper、Swiss Target Prediction平台和Uniprot数据库对靶点补充预测并统一基因名;通过检索GeneCards、OMIM、TTD、CTD、Drugbank、Disgenet和Pharmgkb数据库筛选应激反应调控的相关靶点。使用Venny2.1工具获得参附注射液与应激反应调控交集的潜在作用靶点后,导入STRING数据库构建蛋白相互作用网络并筛选关键靶点。将潜在作用靶点上传至Metascape数据库中,通过GO功能富集分析和KEGG通路富集分析进行机制研究;使用Autoduck、Pymol软件进行分子对接及可视化展示。结果筛选并获取参附注射液主要活性成分43个,相关作用靶点257个;数据库检索到应激反应调控相关靶点4811个,与参附注射液成分相关靶点取交集获得188个潜在作用靶点,通过蛋白相互作用网络筛选得到关键靶点14个。GO功能富集分析筛选得到18条,主要涉及循环系统及体液调控、对外来刺激及创伤的反应、MAPK级联反应、突触后膜、受体复合体、离子通道复合体、神经递质受体活性等。KEGG通路富集分析高度相关通路20条,主要涵盖神经活性配体-受体的相互作用、钙信号通路、肾上腺素能信号转导、类固醇激素生物合成、IL-17、TNF、MAPK、cGMP-PKG、PI3K-Akt、NF-κB、Toll样受体信号通路和细胞凋亡等。分子对接结果提示主要活性成分与关键靶点具有较好的结合力。结论参附注射液中山柰酚、β-谷甾醇、去甲基棱砂贝母碱、豆甾醇、人参皂苷、蛇床子苷、次乌头碱等成分可能作用于AKT1、TNF、IL1B、PTGS2、HSP90AA1、MAPK1、NFKBIA、NR3C1、ADRB2等靶点,通过调节神经-内分泌-免疫系统功能状态、抑制炎症反应、抗氧化应激和减少细胞凋亡等机制对应激反应进行调控。

Objective To investigate the potential mechanism of Shenfu injection in regulating stress response via the neuro-endocrine-immune system by network pharmacology and molecular docking.Methods The main active ingredients and related targets of Shenfu injection were screened using the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform.PharmMapper,Swiss Target Prediction platform and Uniprot database were used to predict the target and unify the gene names.GeneCards,OMIM,TTD,CTD,Drugbank,Disgenet and Pharmgkb databases were searched to screen the related targets regulated by stress responses.Venny 2.1 tool was used to obtain the potential effect targets of the intersection between Shenfu injection and stress response regulation,and the STRING database was imported to construct the interaction PPI network and screen the key targets.Potential effect targets were uploaded to Metascape database online analysis for study on the mechanism through GO and KEGG enrichment analysis.Autoduck and Pymol were used for molecular docking and visualization.Results Forty-three main active ingredients and 257 related targets for Shenfu injection were obtained by component screening and target prediction.A total of 4811 targets related to stress response regulation were retrieved from the database,188 potential effect targets were obtained by intersection with Shenfu injection component-related targets,and 14 key targets were obtained by PPI network screening.Eighteen samples were screened by GO enrichment analysis,which mainly involved the circulatory system and humoral regulation,responses to external stimuli and trauma,MAPK cascade reaction,postsynaptic membrane,receptor complex and ion channel complex and neurotransmitter receptor activity,etc.KEGG enrichment analysis showed 20 highly correlated pathways,mainly covering neuroactive ligand-receptor interaction,calcium signaling pathway,adrenergic signaling,steroid hormone biosynthesis,IL-17,TNF,MAPK,cGMP-PKG,PI3K-Akt,NF-κB,Toll-like receptor signaling pathway and cell apoptosis,etc.The results of molecular docking indicated that the main active components had good binding force with the key target.Conclusions The components of Shenfu injection such as kaempferol,β-sitosterol,Demethyldelavaine,Stigmasterol,ginsenoside,Carnosifloside,hypaconitine may act on targets such as AKT1,TNF,IL1B,PTGS2,HSP90AA1,MAPK1,NFKBIA,NR3C1 and ADRB2 and regulate the stress response through the mechanisms such as regulation of the functional state of the neuro-endocrino-immune system,inhibition of inflammatory responses,anti-oxidative stress and reduction of cell apoptosis.