整合网络药理学和动物实验研究三七皂苷R1抗抑郁的作用与分子机制

Antidepressant effect and molecular mechanism of notoginsenoside R1 based on network pharmacology and animal experiments

该研究拟通过整合网络药理学和动物实验研究三七皂苷R(notoginsenoside R,NGR)治疗抑郁症的作用与分子机制,为进一步深入研究NGR治疗抑郁症的分子机制提供实验依据与理论指导。该研究通过PharmMapper、SwissTargetPrediction、GeneCards等在线分析平台预测NGR与抑郁症相关靶点,获得NGR治疗抑郁的潜在靶点;基于DAVID数据库进行GO功能注释和KEGG富集分析,筛选药物作用于疾病的可能机制;并利用Cytoscape 3.9.0软件构建蛋白-蛋白互作(PPI)网络图,对其进行拓扑学分析,获得核心靶点。通过分子对接检验NGR与核心靶点的结合活性。另外,采用侧脑室注射脂多糖(LPS)的方法制备抑郁模型,给予NGR进行干预,通过行为学测试及RT-qPCR检测NGR抗抑郁作用。结果显示,网络药理学分析可预测56个NGR与抑郁症共同靶点,GO富集分析结果主要涉及G蛋白偶联受体信号通路、RNA聚合酶Ⅱ启动子转录的正调控、细胞因子介导的信号通路、基因表达、凋亡、细胞增殖、信号转导等13个相关生物过程;KEGG富集分析得到神经活性配体-受体相互作用信号通路、脂质和动脉粥样硬化信号通路、cAMP信号通路、PI3K-AKT信号通路等10条潜在作用通路。PPI网络分析预测核心靶点包括CASP3、VEGFA、IGF1、STAT3、MAPK1、PPARG、MTOR、MAPK14、NR3C1、AR等,分子对接结果表明NGR与这些靶点蛋白均具有较好的结合活性。动物实验表明NGR可有效改善抑郁小鼠的疾病行为,并能显著抑制脑部神经炎症指标(Iba-1、TNF-α、IL-1β、IL-6)的mRNA表达,缓解神经炎症反应,同时也可调控脂质和动脉粥样硬化信号通路中的相关靶点(CASP3、STAT3、MAPK1、MAPK14)的mRNA表达水平,说明NGR抗抑郁的作用机制可能与调节脂质和动脉粥样硬化信号通路相关。综上,该研究运用网络药理学预测了NGR的核心靶点和通路并通过动物实验进行了相关的验证,为深入探讨NGR治疗抑郁的作用机制提供依据。

To provide experimental basis and theoretical guidance for further research on the molecular mechanism of notoginsenoside R(NGR)in the treatment of depression,the present study analyzed the potential mechanism of NGRin the treatment of depression through network pharmacology and verified it by molecular docking and animal experiments.PharmMapper,SwissTargetPrediction,and GeneCards were used to predict the related targets of both NGRand depression to obtain the potential targets of NGRin the treatment of depression.The database for annotation,visualization and integrated discovery(DAVID)was used for GO functional annotation and KEGG pathway enrichment analysis to screen the possible mechanisms of NGRexerting antidepressant effect.Cytoscape 3.9.0 was adopted to construct a protein-protein interaction(PPI)network,and the topological analysis was performed to obtain the core targets.The binding activity of NGRto core targets was tested by molecular docking.The depression model was prepared by injecting lipopolysaccharide(LPS)into the lateral ventricle in mice,and intervened with NGR.The antidepressant effect of NGRwas detected by behavioral tests and RT-qPCR.The results showed that by network pharmacology,56 common targets of NGRand depression were predicted,and GO enrichment analysis determined 13 related biological processes,mainly involving G protein-coupled receptor signaling pathway,positive regulation of transcription from RNA polymeraseⅡpromoter,cytokine-mediated signaling pathway,gene expression,apoptosis,cell proliferation,and signal transduction.In addition,KEGG pathway enrichment analysis identified ten potential pathways,including neuroactive ligand-receptor interaction signaling pathway,lipid and atherosclerosis signaling pathway,cAMP signaling pathway,PI3 K-AKT signaling pathway,and lipid and atherosclerosis signaling pathway.PPI analysis revealed that the core targets included CASP3,VEGFA,IGF1,STAT3,MAPK1,PPARG,MTOR,MAPK14,NR3 C1 and AR,and molecular docking demonstrated that NGRhad desirable binding activity to these target proteins.In animal experiments,the results showed that NGRimproved the disease behavior of depressed mice,significantly inhibited the neuroinflammatory response(reducing the mRNA expression of Iba-1,TNF-α,IL-1β,and IL-6),and regulated the mRNA expression of lipid and atherosclerosis signaling pathway-related targets(CASP3,STAT3,MAPK1 and MAPK14).This indicated that the antidepressant mechanism of NGRmay be related to the regulation of lipid and atherosclerosis signaling pathway.In conclusion,network pharmacology was used to reveal the core targets and pathways of NGR,and some of them were verified in animal experiments,which provided the basis for in-depth exploration on the mechanism of NGRin the treatment of depression.