四妙丸（SMP）是一种经典处方，具有抗炎、镇痛、免疫调节等作用，临床上常用于治疗风湿性关节炎（RA）和痛风性关节炎等炎症性疾病，其作用和机制仍不清楚。本研究采用超高效液相色谱-四极杆飞行时间质谱代谢组学技术和液相色谱串联质谱蛋白质组学技术以及网络药理学方法分析RA大鼠血清样品，探索SMP的药效成分。为了进一步验证上述结果，我们构建了类风湿滑膜细胞（FLS）模型，并以黄柏碱进行实验。所有这些线索表明，SMP可以显著降低完全弗氏佐剂大鼠血清中白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的水平，并改善足部肿胀程度；结合代谢组学、蛋白质组学和网络药理学技术，确定了SMP通过炎症途径发挥治疗作用，而黄柏碱是其中一种药效成分。通过构建FLS模型，进一步确定黄柏碱可以有效抑制滑膜细胞活性，通过下调与TLR4-MyD88-IRAK4-MAPK信号通路中相关蛋白的表达水平来减轻关节炎和软骨损伤的炎症水平。总体而言，这些发现表明黄柏碱是SMP治疗RA的有效成分之一。 ©2023 The Authors. Published by American Chemical Society.

The Simiao pill (SMP) is a classic prescription that has shown anti-inflammatory, analgesic, and immunomodulatory effects and is clinically used to treat inflammatory diseases, such as rheumatoid arthritis (RA) and gouty arthritis, for which the effects and mechanism of action remain largely unknown. In this study, serum samples from RA rats were analyzed using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry based metabolomics technology and liquid chromatography with tandem mass spectrometry proteomics technology together with network pharmacology to explore the pharmacodynamic substances of SMP. To further verify the above results, we constructed a fibroblast-like synoviocyte (FLS) cell model and administered phellodendrine for the test. All these clues suggested that SMP can significantly reduce the level of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in complete Freund's adjuvant rat serum and improve the degree of foot swelling; combined with metabolomics, proteomics, and network pharmacological technology, it is determined that SMP plays a therapeutic role through the inflammatory pathway, and phellodendrine is found to be one of the pharmacodynamic substances. By constructing an FLS model, it is further determined that phellodendrine could effectively inhibit the activity of synovial cells and reduce the expression level of inflammatory factors by downregulating the expression level of related proteins in the TLR4-MyD88-IRAK4-MAPK signal pathway to alleviate joint inflammation and cartilage injury. Overall, these findings suggested that phellodendrine is an effective component of SMP in the treatment of RA.© 2023 The Authors. Published by American Chemical Society.