基于网络药理学及体外实验探究黄连阿胶汤治疗抑郁症的机制

Mechanism of Huanglian Ejiao Decoction in the Treatment of Depression Based on Network Pharmacology and in vitro Experiments

目的:探讨黄连阿胶汤治疗抑郁症的机制。方法:通过中药系统药理学数据库与分析平台等对黄连阿胶汤的有效成分及靶点进行筛选,使用Cytoscape软件建立成分-疾病靶点图,基于STRING平台建立交集靶点的蛋白质-蛋白质相互作用(PPI)图,基于DAVID平台进行基因本体(GO)功能富集分析及京都基因与基因组百科全书(KEGG)通路富集分析,基于AutoDock软件进行分子对接。体外实验,取皮质酮造模的PC12细胞,给予槲皮素、黄芩素,检测细胞活性以及相关靶蛋白的表达情况。结果:从黄连阿胶汤中共筛选出抗抑郁的活性成分73种,涉及577个靶点,其中283个靶点与抑郁症相关。槲皮素、黄芩素和β-谷甾醇等活性成分可能会发挥重要作用。PPI结果显示,信号转导及转录激活因子3、血管内皮生长因子A、白细胞介素(IL)1β、IL-6、肿瘤坏死因子(TNF)和蛋白激酶B1是黄连阿胶汤治疗抑郁症的核心靶点。分子对接结果显示,靶点与有效成分之间有着良好的结合能力。GO和KEGG富集分析结果显示,283个靶点涉及缺氧反应、突触后膜和质膜,以及缺氧诱导因子(HIF)1、TNF和环磷酸腺苷(cAMP)等信号通路。体外实验结果表明,槲皮素、黄芩素可以有效改善皮质酮对PC12细胞的凋亡率,降低炎症因子的表达等。结论:经过网络药理学的预测及体外实验的验证,黄连阿胶汤可通过HIF-1、TNF和cAMP等信号通路调节炎症因子、细胞因子及氧化应激反应等治疗抑郁症。

OBJECTIVE:To explore the mechanism of Huanglian Ejiao decoction in the treatment of depression.METHODS:Effective components and targets of Huanglian Ejiao decoction were screened through Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform and other database.Component-disease-target network was established by Cytoscape.Protein-protein interaction(PPI)of intersection targets was established based on STRING platform.Gene ontology(GO)functional enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were performed based on DAVID platform.AutoDock software was used for the molecular docking.In vitro experiments,the PC12 cell model was administered with quercetin and baicalein,cell activity and expression of related target proteins were detected.RESULTS:A total of 73 antidepressant active ingredients were screened from Huanglian Ejiao decoction,including 577 targets,of which 283 targets were related to depression.Active ingredients such as quercetin,baicalein and beta-sitosterol may play an important role.PPI results showed that signal transducer and activator of transcription 3,vascular endothelial growth factor A,interleukin(IL)1β,IL-6,tumor necrosis factor(TNF)and protein kinase B1 were the core targets of Huanglian Ejiao decoction in the treatment of depression.Molecular docking results showed that there was a good binding ability.GO and KEGG enrichment analysis showed that 283 targets were involved in hypoxia response,postsynaptic membrane and plasma membrane,as well as signal pathways such as hypoxia inducible factor(HIF)1,TNF and cyclic adenosine monophosphate(cAMP).Results of in vitro experiments showed that quercetin and baicalein could effectively improve the apoptosis rate of PC12 cells and reduce the expression of inflammatory factors.CONCLUSIONS:After the prediction of network pharmacology and verification of in vitro experiments,Huanglian Ejiao decoction can treat depression by regulating inflammatory factors,cytokines and oxidative stress through HIF-1,TNF,cAMP and other signaling pathways.