基于网络药理学探讨干姜附子汤抗心肌缺血再灌注损伤的作用机制

Effect of dried ginger-aconite decoction on myocardial ischemia-reperfusion injury and related mechanism based on network pharmacology

目的基于网络药理学探讨干姜附子汤抗心肌缺血再灌注损伤(MI/RI)的作用机制。方法通过TCMSP数据库筛选检索干姜、附子的主要成分,并通过TCMSP和CTD数据库筛选成分靶点,结合Disgenet数据库获取干姜附子汤抗MI/RI的作用靶点。应用STRING平台构建蛋白-蛋白相互作用(PPI)网络模型,应用DAVID数据库进行GO富集分析和KEGG通路富集分析,进一步进行实验验证,以揭示干姜附子汤抗MI/RI的机制。结果筛选得到16个活性成分,171个靶点,治疗MI/RI靶点80个。GO功能富集分析得到GO条目158个,包括生物过程条目118个,细胞组成条目22个,分子功能条目18个。KEGG通路富集筛选得到102条信号通路,主要涉及PI3K-Akt、TNF、HIF-1、甲状腺激素、细胞凋亡等信号通路。流式细胞仪检测结果显示干姜附子汤可减少缺氧损伤的H9C2细胞凋亡率。试剂盒检测Na+-K+-ATP、Ca2+-Mg2+-ATP酶活力,发现干姜附子汤可显著提高Na+-K+-ATP、Ca2+-Mg2+-ATP酶活性。结论干姜附子汤治疗MI/RI呈现多分子、多靶点、多通路的协同作用机制,且治疗机制与凋亡密切相关。

Objective To determine the potential molecular mechanism of dried ginger-aconite decoction in treating myocardial ischemia-reperfusion injury (MI/RI).Methods The main components of dried ginger and aconite were retrieved and screened through the TCMSP database,and the component targets were screened through the TCMSP and CTD database.Combined with the Disgenet database,the therapeutic targets of dried ginger-aconite decoction for MI/RI were obtained.The STRING platform was used to construct a protein-protein interaction (PPI) network model.DAVID database was used for GO enrichment analysis and KEGG pathway enrichment analysis,and further experiments were conducted to verify the mechanism of dried ginger-aconite decoction for MI/RI.Results Totally 16 active components,171 targets,and 80 MI/RI targets were screened.GO functional enrichment analysis yielded 158 GO entries,including 118 biological process entries,22 cell composition entries,and 18 molecular function entries.The enrichment and screening of KEGG pathway resulted in 102 signaling pathways,mainly involving PI3K-Akt,TNF,HIF-1,and thyroid hormone,apoptosis and other signaling pathways.The activities of Na+-K+-ATPase and Ca~2+-Mg~2+-ATPase were detected by kit.Dried ginger-aconite decoction was found to significantly increase the activities of Na+-K+-ATP and Ca~2+-Mg~2+-ATPase.The flow cytometry showed that dried ginger-aconite decoction reduced the apoptosis rate of H9C2 cells damaged by hypoxia reoxygenation.Conclusion Dried ginger-aconite decoction in the treatment of MI/RI presents a multi-molecule,multi-target,and multi-channel synergistic mechanism and the therapeutic mechanism is closely related to apoptosis.