目的 利用网络药理学、分子对接技术，从肾茶中筛选潜在的治疗痛风性关节炎的活性成分及其靶点，并通过体外实验进一步验证。方法 从中药系统药理学数据库与分析平台（TCMSP）、HERB等数据库获得肾茶的活性成分及其作用靶点，通过人类基因数据库（GeneCards）、人类孟德尔遗传数据库（OMIM）等获得痛风性关节炎的相关靶点，采用R软件的Venny 2.1.0包获取肾茶与痛风性关节炎的交集靶点，利用软件CytoScape 3.9.1绘制“肾茶–活性成分–疾病靶点”网络，通过STRING网站构建关键靶点的蛋白质–蛋白质相互作用（PPI）网络，并通过微生信平台进行基因本体（GO）功能和京都基因与基因组百科全书（KEGG）通路富集分析。利用分子对接技术对核心活性成分和核心靶点对接结果进行模拟可视化。使用脂多糖（LPS）诱导RAW264.7巨噬细胞，构建体外炎症模型，给药组则同时予以黄芩素（5、10、20 μmol/L）、甜橙黄酮（6.25、12.5、25 μmol/L）和鼠尾草素（12.5、25、50 μmol/L）处理。通过RT-qPCR检测白细胞介素-6（IL-6）、IL-1β、肿瘤坏死因子-α（TNF-α）mRNA表达水平和Western blotting检测胞外信号调节激酶（ERK）、核因子-κB（NF-κB）p65磷酸化水平。结果 网络药理学最终筛选得到7个肾茶活性成分，对应靶点481个，与痛风性关节炎交集靶点共47个；“肾茶–活性成分–疾病靶点”网络识别出3种核心成分（黄芩素、甜橙黄酮和鼠尾草素）；PPI筛选得到NOD样受体热蛋白结构域相关蛋白3（NLRP3）、肉瘤病毒癌基因同源物（SRC）、集落刺激因子1受体（CSF1R）、基质金属蛋白酶9（MMP9）、肿瘤坏死因子受体超家族成员1A（TNFRSF1A）、白细胞介素-18（IL18）、白细胞介素-10（IL10）、干扰素-γ（IFNG）、Toll样受体4（TLR4）、丝裂原活化蛋白激酶14（MAPK14）10个核心靶点；GO和KEGG通路分析可知肾茶通过MAPK及NF-κB等信号通路发挥作用；分子对接结果显示，3种核心成分与痛风性关节炎治疗靶点的结合能力各有不同，与MAPK14结合能力较强。RT-qPCR实验结果显示，给药组较模型组IL-6、IL-1β、TNF-α mRNA表达水平明显下降；Western blotting实验结果显示，与模型组相比，给药后抑制细胞中MAPK通路及NF-κB通路的激活，显著降低ERK、NF-κB p65磷酸化水平。结论 肾茶治疗痛风性关节炎的主要活性成分为黄芩素、甜橙黄酮、鼠尾草素，以及药物主要通过MAPK/ERK和NF-κB p65信号通路发挥治疗痛风性关节炎作用。

Objective To screen the potential active ingredients and targets of Clerodendranthus spicatus against gouty arthritis based on network pharmacology and molecular docking technology, and to further validate the findings through in vitro experiments. Methods The active ingredients of C. spicatus and their corresponding targets were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and the HERB database. Disease targets of gouty arthritis were retrieved from the GeneCards database and the Online Mendelian Inheritance in Man (OMIM) database. Intersection targets between C. spicatus and gouty arthritis were identified using the Venny 2.1.0 package in R software. The “C. spicatus-active component-disease target” network was constructed using Cytoscape 3.9.1. A protein-protein interaction (PPI) network of core targets was built via the STRING database. Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed using the Bioinformatics Platform. Molecular docking was employed to simulate and visualize the binding of core active components to core targets. An in vitro inflammatory model was established using lipopolysaccharide (LPS)-induced RAW264.7 macrophages. Treatment groups were administered baicalein (5, 10, 20 μmol/L), sinensetin (6.25, 12.5, 25 μmol/L), and salvigenin (12.5, 25, 50 μmol/L). The mRNA expression levels of interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α) were measured by RT-qPCR. The phosphorylation levels of extracellular signal-regulated kinase (ERK) and nuclear factor-κB (NF-κB) p65 were detected by Western blotting. Results Network pharmacology ultimately screened 7 active components of C. spicatus, corresponding to 481 targets, and 47 intersection targets with gouty arthritis. Three core components (baicalein, sinensetin, and salvigenin) were identified from the “C. spicatus-active component-disease target” network. PPI network analysis yielded 10 core targets: NOD-like receptor thermal protein domain associated protein 3 (NLRP3), sarcoma virus oncogene homolog (SRC), colony-stimulating factor 1 receptor (CSF1R), matrix metalloproteinase 9 (MMP9), tumor necrosis factor receptor superfamily member 1A (TNFRSF1A), interleukin-18 (IL18), interleukin-10 (IL10), interferon-γ (IFNG), Toll-like receptor 4 (TLR4), and mitogen-activated protein kinase 14 (MAPK14). GO and KEGG pathway analyses indicated that C. spicatus exerts its effects through signaling pathways such as the MAPK and NF-κB pathways. Molecular docking results showed that the three core components each have varying binding affinities to gouty arthritis therapeutic targets, with relatively strong binding to MAPK14. RT-qPCR results demonstrated that compared with the model group, the treatment groups significantly reduced the mRNA expression levels of IL-6, IL-1β, and TNF-α. Western blotting results showed that compared with the model group, treatment inhibited the activation of the MAPK and NF-κB pathways in cells and significantly reduced the phosphorylation levels of ERK and NF-κB p65. Conclusion The main active ingredients of C. spicatus against gouty arthritis are baicalein, sinensetin, and salvigenin, and the therapeutic effect is primarily mediated through the MAPK/ERK and NF-κB p65 signaling pathways.

江西中医药大学中药学（新药创制方向）特区建设立项项目（TQ-11）；江西中医药大学中药药效物质基础江西省重点实验室项目（2024SSY07102）；江西中医药大学校企合作项目（横20220124）