目的 基于网络药理学和分子对接技术结合体内实验验证，探讨归芍地黄汤加减方治疗抽动秽语综合征（Tourette syndrome，TS）的作用机制。方法 通过检索TCMSP和BATMAN-TCM数据库获得归芍地黄汤加减方的活性成分及作用靶点，检索GeneCards和OMIM数据库获得TS疾病相关靶点，将二者靶点取交集，得到交集靶点。利用Cytoscape软件构建活性成分-靶点网络，并对交集靶点进行蛋白质-蛋白质相互作用（protein-protein interaction，PPI）及网络拓扑分析。利用R语言软件及相关程序包对交集靶点进行基因本体（gene ontology，GO）功能及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析，对网络拓扑分析后筛选出的关键靶点进行分子对接，并通过动物实验对关键靶点和信号通路进行验证。结果 筛选得到归芍地黄汤加减方治疗TS的活性成分8个，交集靶点32个。主要活性成分包括槲皮素、豆甾醇、β-谷甾醇、异鼠李素，关键靶点包括白细胞介素-1β（interleukin-1β，IL-1β）、溶质载体家族6成员4（solute carrier family 6 member 4，SLC6A4）、单胺氧化酶B（monoamine oxidase B，MAOB）、CXC趋化因子配体8（C-X-C motif chemokine ligand 8，CXCL8）、趋化因子配体2（C-C motif chemokine ligand 2，CCL2）、单胺氧化酶A（monoamine oxidase A，MAOA）、骨肉瘤癌基因（FBJ osteosarcoma oncogene，FOS）。GO功能富集分析表明归芍地黄汤加减方可能通过儿茶酚胺结合、G蛋白偶联胺受体活性、铵离子结合、神经递质受体活性等治疗TS。KEGG通路富集分析表明归芍地黄汤加减方可能通过药物代谢-细胞色素P450、神经活性配体-受体相互作用、细胞因子-细胞因子-受体相互作用、IL-17信号通路、核因子-κB（nuclear factor-κB，NF-κB）信号通路等治疗TS。经归芍地黄汤加减方干预后，TS大鼠抽动频率、刻板运动和临床症状均显著改善（P<0.05），血清中炎症因子IL-1β、CXCL8、CCL2水平均显著降低（P<0.05、0.01），纹状体中NF-κB p65、p-NF-κB p65、kappa B抑制蛋白激酶（inhibitor of kappa B kinase，IKK）蛋白表达水平均显著降低（P<0.05、0.01），NF-κB抑制蛋白（inhibitor of NF-κB，IκB）蛋白表达水平显著升高（P<0.01）。结论 归芍地黄汤加减方通过多组分多靶点发挥治疗TS的作用，其作用机制可能与调控NF-κB信号通路、改善神经炎症相关。

Objective To explore the mechanism of modified Guishao Dihuang Decoction (归芍地黄汤加减方) on Tourette syndrome (TS) based on network pharmacology and molecular docking combined with in vivo experimental verification. Methods The active constituents and targets of modified Guishao Dihuang Decoction were identified through TCMSP and BATMAN-TCM databases. Targets of TS were retrieved from GeneCards and OMIM database, the two targets were intersected to obtain the interleaved target. Active ingredient-target network was constructed by Cytoscape software, followed by protein-protein interaction (PPI) and network topology analysis for the interleaved targets. For the interleaved targets, gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were conducted using R language software and relevant programs. Additionally, molecular docking was performed for key targets selected based on the results of network topology analysis. The key targets and signaling pathway were verified by animal experiments. Results A total of eight active components and 32 interleaved targets of modified Guishao Dihuang Decoction were selected for TS treatment. Major active ingredients included quercetin, stigmasterol, beta-sitosterol, isorhamnetin and others. Key targets included interleukin-1β (IL-1β), solute carrier family 6 member 4 (SLC6A4), monoamine oxidase B (MAOB), C-X-C motif chemokine ligand 8 (CXCL8), C-C motif chemokine ligand 2 (CCL2), monoamine oxidase A (MAOA) and FBJ osteosarcoma oncogene (FOS). GO function enrichment analysis suggested that modified Guishao Dihuang Decoction may exert an effect on TS by catecholamine binding, G protein-coupled amine receptor activity, ammonium ion binding, neurotransmitter receptor activity, etc. KEGG pathway enrichment analysis indicated that modified Guishao Dihuang Decoction could play its effects through drug metabolism-cytochrome P450, neuroactive ligand-receptor interaction, cytokine-cytokine-receptor interaction, IL-17 signaling pathway, nuclear factor-κB (NF-κB) signaling pathway, etc. After the intervention of modified Guishao Dihuang Decoction, the tic frequency, rigid movement and clinical symptoms were significantly improved (P < 0.05), levels of inflammatory factors IL-1β, CXCL8 and CCL2 in serum were significantly decreased (P < 0.05, 0.01), NF-κB p65, p-NF-κB p65 and inhibitor of kappa B kinase (IKK) protein expression levels in striatum were significantly decreased (P < 0.05, 0.01), while inhibitor of NF-κB (IκB) protein expression level was significantly increased (P < 0.01). Conclusions Modified Guishao Dihuang Decoction plays a role in treatment of TS through multi-components and multi-targets, and its mechanism may be related to regulating NF-κB signaling pathway and improving neuroinflammation.

R285.5

广州华商学院校内学术科研项目（2023HSXS01）