Shi Wei Ru Xiang pill alleviates acute gouty arthritis through suppressing NLRP3 inflammasome activation

Shi Wei Ru Xiang pill (SWR) is commonly utilized in Tibetan medicine as a therapeutic intervention for 'Huang-shui disease' and has been clinically validated as an effective treatment for acute gouty arthritis (AGA). Nevertheless, the underlying mechanisms of action and the active components of SWR in combating AGA remain unclear. In this study, the effects of SWR and its active components on AGA and NLRP3 inflammasome activation were investigated in monosodium urate (MSU)-induced AGA rats and lipopolysaccharide/nigericin-induced THP-1 cells. The chemical profile of SWR was characterized using UPLC-Q-Exactive Orbitrap MS. The results indicated that SWR effectively suppressed pyroptosis, caspase-1 activity, and IL-1β production in THP-1 cells. Furthermore, SWR significantly suppressed NLRP3 inflammasome activation and attenuated ankle swelling in a rat AGA model. Specifically, SWR affected the priming and assembly phases to inhibit NLRP3 inflammasome activation. Surprisingly, SWR showed good liver and renal protective effects in AGA rats. A total of 58 compounds were identified in SWR by UPLC-MS analysis. Further pharmacological studies demonstrated that the phenolic compounds serve as active compounds responsible for the inhibition of inflammasome activation, including dehydrocostus lactone, gallic acid, 4-hydroxybenzoic acid. This is the first study to comprehensively elucidate the therapeutic effects and underlying mechanisms of SWR against AGA by inhibiting NLRP3 inflammasome activation. This study strongly indicated that SWR could serve as a promising anti-inflammatory medicine with an acceptable safety profile for the treatment of AGA and other inflammatory disorders linked to NLRP3 inflammasome activation十味乳香丸（Shi Wei Ru Xiang pill, SWR）是藏医药中常用于治疗黄水病（Huang-shui disease）的治疗方剂，且经临床验证可有效治疗急性痛风性关节炎（acute gouty arthritis, AGA）。然而，十味乳香丸对抗急性痛风性关节炎的潜在作用机制及其活性成分仍不明确。本研究分别在尿酸钠（monosodium urate, MSU）诱导的急性痛风性关节炎模型大鼠，以及脂多糖/尼日利亚菌素诱导的THP-1细胞中，探究了十味乳香丸及其活性成分对急性痛风性关节炎与NLRP3炎性小体（NLRP3 inflammasome）活化的影响。采用超高效液相色谱-静电场轨道阱质谱（UPLC-Q-Exactive Orbitrap MS）对十味乳香丸的化学组分进行了表征。结果显示，十味乳香丸可有效抑制THP-1细胞的细胞焦亡、半胱天冬氨酸蛋白酶-1（caspase-1）活性以及白细胞介素-1β（IL-1β）的产生。此外，十味乳香丸可显著抑制NLRP3炎性小体活化，并减轻急性痛风性关节炎模型大鼠的踝关节肿胀症状。具体而言，十味乳香丸通过影响炎性小体的启动与组装阶段，从而抑制NLRP3炎性小体活化。值得注意的是，十味乳香丸对急性痛风性关节炎模型大鼠展现出良好的肝、肾保护作用。经超高效液相色谱-质谱分析，本研究共从十味乳香丸中鉴定出58种化合物。进一步的药理学研究证实，酚类化合物为其抑制炎性小体活化的活性成分，包括去氢木香内酯、没食子酸及4-羟基苯甲酸。本研究为首个通过抑制NLRP3炎性小体活化，全面阐明十味乳香丸抗急性痛风性关节炎的疗效与潜在作用机制的研究。本研究结果表明，十味乳香丸有望成为治疗急性痛风性关节炎及其他与NLRP3炎性小体活化相关炎症性疾病的极具潜力的抗炎药物，且安全性良好。