目的 应用网络药理学分析金藤清痹颗粒治疗急性痛风性关节炎（acute gouty arthritis，AGA）的作用机制，并建立AGA大鼠模型进行验证。方法 在清热解毒、活血止痛治法指导下，通过TCMSP数据库搜集金藤清痹颗粒的活性成分及靶点，利用GeneCards、NCBI数据库等搜集AGA相关靶点，与金藤清痹颗粒作用靶点整合后，构建共有靶点蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络和“金藤清痹颗粒-中药-活性成分-靶点-AGA”网络，并进行基因本体（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。雄性SD大鼠随机分为空白组、模型组、秋水仙碱（0.3 mg/kg）组和金藤清痹颗粒低、中、高剂量（1.05、2.10、4.20 g/kg）组，每组6只，踝关节注射单钠尿酸盐（monosodium urate，MSU）晶体建立AGA大鼠模型。采用游标卡尺测量大鼠踝关节直径，计算踝关节肿胀度；采用全自动生化仪检测血清尿酸（serum uric acid，SUA）、C反应蛋白（C-reactive protein，CRP）水平；采用苏木素-伊红（HE）染色观察踝关节组织病理变化；采用qRT-PCR、ELISA和Western blotting检测踝关节组织及血清中关键靶点和信号通路的表达。结果 共检索到金藤清痹颗粒110种活性成分、212个作用靶点，272个AGA治疗靶点，共有靶点29个，关键靶点有白细胞介素-1β（interleukin-1β，IL-1β）、肿瘤坏死因子（tumor necrosis factor，TNF）及IL-6，涉及TNF信号通路、IL-17信号通路和NOD样受体信号通路等。大鼠踝关节注射MSU晶体后明显肿胀（P<0.01），SUA及CRP显著升高（P<0.05、0.01），滑膜组织增生明显、结构紊乱，有大量炎症细胞浸润，NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor thermal protein domain associated protein 3，NLRP3）、NIMA相关蛋白激酶7（NIMA-related kinases 7，NEK7）、凋亡相关斑点样蛋白（apoptosis-associated speck-like protein containing a CARD，ASC）、半胱氨酸天冬氨酸蛋白酶-1（cystein-asparate protease-1，Caspase-1）、消皮素D（gasdermin D，GSDMD）、IL-18、IL-1β、IL-6及TNF-α表达水平明显升高（P<0.01）；秋水仙碱或金藤清痹颗粒治疗后，有效缓解踝关节肿胀（P<0.05、0.01），SUA及CRP均显著降低（P<0.05、0.01），关节滑膜增生、炎症细胞浸润均得到改善，同时显著抑制IL-1β、TNF-α及IL-6等靶点和NOD样受体信号通路的表达（P<0.05、0.01）。结论 金藤清痹颗粒对AGA大鼠具有明显的保护作用，其机制可能与抑制NOD样受体信号通路的异常激活，降低炎症因子水平，改善关节滑膜增生、炎症细胞浸润有关。

Objective To study the mechanism of Jinteng Qingbi Granules (金藤清痹颗粒) in treating acute gouty arthritis (AGA) by network pharmacology and establish AGA rat model for verification. Methods The active components and targets of Jinteng Qingbi Granules were collected by TCMSP database, and AGA-related targets were collected by GeneCards and NCBI database under the guidance of clearing heat and detoxification, promoting blood circulation and relieving pain. After integrating with the targets of Jinteng Qingbi Granules, protein-protein interaction (PPI) and "Jinteng Qingbi Granules-traditional Chinese medicine-active component-target-AGA" network was constructed, gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were carried out. Male SD rats were randomly divided into blank group, model group, colchicine (0.3 mg/kg) group and Jinteng Qingbi Granules low-, medium-and high-dose (1.05, 2.10, 4.20 g/kg) groups, with six rats in each group. The AGA rat model was established by ankle injection of monosodium urate (MSU) crystals. The diameter of ankle joint in rats was measured by vernier caliper, and the swelling degree of ankle joint was calculated. Serum uric acid (SUA) and C-reactive protein (CRP) levels were detected by automatic biochemical analyzer. HE staining was used to observe the pathological changes of ankle joint. qRT-PCR, ELISA and Western blotting were used to detect the expressions of key targets and signal pathways in ankle joint tissue and serum. Results A total of 110 active ingredients, 212 targets and 272 AGA therapeutic targets were retrieved, with a total of 29 common targets. The key targets were interleukin-1β (IL-1β), tumor necrosis factor (TNF) and IL-6, involving TNF signaling pathway, IL-17 signaling pathway and NOD-like receptor signaling pathway. After injecting MSU crystal into the ankle joint of rats, the swelling was obvious (P < 0.01), SUA and CRP were significantly increased (P < 0.05, 0.01), synovial tissue proliferated obviously, the structure was disordered, and a large number of inflammatory cells infiltrated. NOD-like receptor thermal protein domain associated protein 3 (NLRP3), NIMA-related kinases 7 (NEK7), apoptosis-associated speck-like protein containing a CARD (ASC), cystein-aspartate protease-1 (Caspase-1), gasdermin D (GSDMD), IL-18, IL-1β, IL-6 and TNF-α expression levels were significantly increased (P < 0.01). After treatment with colchicine or Jinteng Qingbi Granules, ankle swelling was effectively relieved (P < 0.05, 0.01), SUA and CRP were significantly decreased (P < 0.05, 0.01), synovial hyperplasia and inflammatory cell infiltration of joints were improved, and the expressions of IL-1β, TNF-α and IL-6 and NOD-like receptor signaling pathway were significantly inhibited (P < 0.05, 0.01). Conclusion Jinteng Qingbi Granules has significant protective effects on AGA rats, and the mechanism may be related to the inhibition of abnormal activation of NOD-like receptor signaling pathway, reducing inflammatory factor levels, and improvement of synovial hyperplasia and inflammatory cell infiltration in joints.

R285.5

国家自然科学基金资助项目（82074377）；中医药传承与创新“百千万”人才工程（岐黄工程）岐黄学者（20210602-1）；国家中医药管理局名老中医传承工作室（975022）；金藤清痹颗粒二次开发项目（20220626-1）