目的 优化化湿止痛方的提取工艺与凝胶剂制备工艺，探究化湿止痛凝胶对痛风性关节炎的治疗效果及作用机制，为痛风性关节炎外用治疗新药研发提供候选制剂与理论依据。方法 采用紫外分光光度法测定化湿止痛方的总酚酸含量，通过单因素与正交试验优化提取工艺；以外观、黏度、延展性及稳定性为指标筛选凝胶基质及辅料配比，运用Box-Behnken响应面法优化制备工艺。建立大鼠急性痛风性关节炎模型，通过检测关节肿胀、皮温、疼痛及炎症因子[白细胞介素-1β（interleukin-1β，IL-1β）、巨噬细胞炎症蛋白-1α（macrophage inflammatory protein-1α，MIP-1α）、超氧化物歧化酶（superoxide dismutase，SOD）、Toll样受体4（Toll-like receptor 4，TLR4）、基质金属蛋白酶9（matrix metalloproteinase 9，MMP9）]表达，考察化湿止痛凝胶抗痛风性关节炎的药效作用。结合GEO数据、网络药理学与分子对接构建“物质-效应组群”网络，分析化湿止痛凝胶抗痛风性关节炎的作用机制并进行实验验证。结果 化湿止痛方的最佳提取工艺为60%乙醇、料液比1∶10、70 ℃提取1.5 h，提取2次；凝胶剂最佳配方为1%卡波姆940与0.2%泊洛沙姆188为基质，搭配7.5%保湿剂、1.2%促渗剂、14.7%含药量及0.07%抗氧剂。药效学实验结果显示，化湿止痛凝胶能显著减轻痛风性关节炎大鼠关节肿胀和疼痛（P＜0.01、0.001），降低血清中IL-1β、MIP-1α水平及滑膜组织TLR4、MMP9表达（P＜0.05、0.01），升高血清中SOD活性（P＜0.05、0.01）。机制研究表明，化湿止痛凝胶通过调控前列腺素内过氧化物合酶2（prostaglandin-endoperoxide synthase 2，PTGS2）、表皮生长因子受体（epidermal growth factor receptor，EGFR）、血管内皮生长因子A（vascular endothelial growth factor A，VEGFA）等靶点及环磷酸腺苷（cyclic adenosine monophosphate，cAMP）、趋化因子、钙信号等通路发挥抗痛风性关节炎的作用；免疫荧光染色和qRT-PCR实验证实化湿止痛凝胶能显著抑制痛风性关节炎大鼠滑膜组织环氧化酶-2（cyclooxygenase-2，COX-2）、VEGFA和EGFR的表达（P＜0.01）。结论 成功优化化湿止痛凝胶剂的提取与制备工艺，化湿止痛凝胶具有良好的抗痛风性关节炎作用，且其作用机制与多靶点、多通路调控相关。

Objective To optimize the extraction process of Huashi Zhitong Formula (化湿止痛方) and preparation technology of its gel formulation, elucidate the therapeutic efficacy and mechanism of Huashi Zhitong Gel against gouty arthritis, and provide a candidate formulation and theoretical basis for the development of new topical drugs for gouty arthritis. Methods The total phenolic acid content in Huashi Zhitong Formula was determined by ultraviolet spectrophotometry. The extraction process was optimized through single-factor and orthogonal tests. The gel matrix and excipient ratios were screened based on appearance, viscosity, ductility and stability, and the preparation process was further optimized using Box-Behnken response surface methodology. A rat gouty arthritis model was established, anti-gouty arthritis efficacy of Huashi Zhitong Gel was investigated by detecting joint swelling, skin temperature, pain and expressions of inflammatory factors [interleukin-1β (IL-1β), macrophage inflammatory protein-1α (MIP-1α), superoxide dismutase (SOD), Toll-like receptor 4 (TLR4), matrix metalloproteinase 9 (MMP9)]. Integrated with GEO data, network pharmacology and molecular docking, a “component-effect group” network was constructed to analyze the mechanism of Huashi Zhitong Gel against gouty arthritis, followed by experimental validation. Results The optimal extraction conditions of Huashi Zhitong Formula were determined as 60% ethanol, a solid-to-liquid ratio of 1∶10, extraction at 70 ℃ for 1.5 h (repeated twice). The optimal gel formulation consisted of 1% carbomer 940 and 0.2% poloxamer 188 as the matrix, combined with 7.5% humectant, 1.2% penetration enhancer, 14.7% drug load and 0.07% antioxidant. Pharmacodynamic experiment results demonstrated that Huashi Zhitong Gel significantly alleviated joint swelling and pain in rats with gouty arthritis (P < 0.01, 0.001), reduced the levels of IL-1β, MIP-1α in serum and expressions of TLR4, MMP9 in synovial tissue (P < 0.05, 0.01), and increased the activity of SOD in serum (P < 0.05, 0.01). The mechanism study showed that Huashi Zhitong Gel could play an anti-gouty arthritis role by regulating the targets such as prostaglandin endoperoxide synthase 2 (PTGS2), epidermal growth factor receptor (EGFR), vascular endothelial growth factor A (VEGFA), and the pathways such as cyclic adenosine monophosphate (cAMP), chemokines, and calcium signaling. Immunofluorescence staining and qRT-PCR confirmed that Huashi Zhitong Gel could significantly inhibit the expressions of cyclooxygenase-2 (COX-2), VEGFA and EGFR in synovium of rats with gouty arthritis (P < 0.01). Conclusion The extraction and preparation process of Huashi Zhitong Gel are successfully optimized. Huashi Zhitong Gel has a good anti-gouty arthritis effect, and its mechanism is related to multi target and multi pathway regulation.

R285.5

广西中药药效研究重点实验室项目（桂科计字[2005]6号）；中药资源循环利用广西高校工程研究中心（桂教科研[2024]10号）；广西“带土移植”人才引育计划项目（桂科AA23026008）