Li Ye, Zhang Yan-Yan, Wu Di, Qi Ke-Ming, Lu Wei, Wei Yi-Cong
School of Pharmacy, Fujian University of Traditional Chinese Medicine Fuzhou 350122, China.
Zhongguo Zhong Yao Za Zhi. 2022 Oct;47(20):5599-5609. doi: 10.19540/j.cnki.cjcmm.20220509.704.
To provide experimental basis and theoretical guidance for further research on the molecular mechanism of notoginsenoside R_1(NGR_1) in the treatment of depression, the present study analyzed the potential mechanism of NGR_1 in 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 NGR_1 and depression to obtain the potential targets of NGR_1 in 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 NGR_1 exerting 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 NGR_1 to 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_1. The antidepressant effect of NGR_1 was detected by behavioral tests and RT-qPCR. The results showed that by network pharmacology, 56 common targets of NGR_1 and 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 polymerase Ⅱ promoter, 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 NGR_1 had desirable binding activity to these target proteins. In animal experiments, the results showed that NGR_1 improved 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 NGR_1 may 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_1, and some of them were verified in animal experiments, which provided the basis for in-depth exploration on the mechanism of NGR_1 in the treatment of depression.
为进一步研究三七皂苷R_1（NGR_1）治疗抑郁症的分子机制提供实验依据和理论指导，本研究通过网络药理学分析NGR_1治疗抑郁症的潜在机制，并通过分子对接和动物实验进行验证。利用PharmMapper、SwissTargetPrediction和GeneCards预测NGR_1和抑郁症的相关靶点，以获得NGR_1治疗抑郁症的潜在靶点。使用注释、可视化和综合发现数据库（DAVID）进行基因本体（GO）功能注释和京都基因与基因组百科全书（KEGG）通路富集分析，以筛选NGR_1发挥抗抑郁作用的可能机制。采用Cytoscape 3.9.0构建蛋白质-蛋白质相互作用（PPI）网络，并进行拓扑分析以获得核心靶点。通过分子对接检测NGR_1与核心靶点的结合活性。通过向小鼠侧脑室注射脂多糖（LPS）制备抑郁症模型，并用NGR_1进行干预。通过行为测试和实时定量聚合酶链反应（RT-qPCR）检测NGR_1的抗抑郁作用。结果表明，通过网络药理学预测了NGR_1和抑郁症的56个共同靶点，GO富集分析确定了13个相关生物学过程，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、细胞增殖和信号转导。此外，KEGG通路富集分析确定了10条潜在通路，包括神经活性配体-受体相互作用信号通路、脂质与动脉粥样硬化信号通路、环磷酸腺苷（cAMP）信号通路、磷脂酰肌醇-3激酶/蛋白激酶B（PI3K-AKT）信号通路和脂质与动脉粥样硬化信号通路。PPI分析显示核心靶点包括半胱天冬酶3（CASP3）、血管内皮生长因子A（VEGFA）、胰岛素样生长因子1（IGF1）、信号转导和转录激活因子3（STAT3）、丝裂原活化蛋白激酶1（MAPK1）、过氧化物酶体增殖物激活受体γ（PPARG）、哺乳动物雷帕霉素靶蛋白（MTOR）、丝裂原活化蛋白激酶14（MAPK14）、核受体亚家族3成员C1（NR3C1）和雄激素受体（AR），分子对接表明NGR_1与这些靶蛋白具有良好的结合活性。在动物实验中，结果表明NGR_1改善了抑郁小鼠的疾病行为，显著抑制神经炎症反应（降低离子钙接头蛋白1（Iba-1）、肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）的mRNA表达），并调节脂质与动脉粥样硬化信号通路相关靶点（CASP3、STAT3、MAPK1和MAPK14）的mRNA表达。这表明NGR_1的抗抑郁机制可能与脂质与动脉粥样硬化信号通路的调节有关。总之，利用网络药理学揭示了NGR_1的核心靶点和通路，其中一些在动物实验中得到验证，为深入探索NGR_1治疗抑郁症的机制提供了依据。
