类风湿性关节炎（RA）是一种以滑膜炎和炎性细胞浸润为特征的自身免疫性疾病。中药痹痛清（BTQ）在RA临床治疗中显示出显着疗效。然而，BTQ 治疗 RA 的潜在治疗机制尚未得到充分研究。本研究旨在阐明BTQ对胶原诱导性关节炎(CIA)大鼠巨噬细胞焦亡的影响，为治疗RA提供理论依据。本研究采用液相色谱-质谱联用技术(LC-MS)对BTQ的主要成分进行鉴定。在 CIA 大鼠模型中评估 BTQ 的治疗效果。使用病理组织学染色、免疫荧光、显微 CT 和蛋白质印迹进行体内实验。接下来，使用脂多糖（LPS）和三磷酸腺苷（ATP）诱导小鼠单核巨噬细胞（RAW264.7）发生焦亡，并通过细胞活力、免疫荧光分析评估BTQ对RAW264.7巨噬细胞的影响， BTQ对CIA大鼠有治疗作用，主要表现为关节炎症减轻、足部肿胀、骨侵蚀减轻、病理改变改善。进一步的研究表明，BTQ 抑制细胞因子产生白细胞介素 18 (IL-18) 和白细胞介素 1β (IL-1β) 的水平，同样，它抑制 NOD 样受体热蛋白域相关蛋白中关键蛋白的表达3 (NLRP3) 介导 CIA 大鼠滑膜组织焦亡。体外实验结果表明，BTQ 可减弱 LDH 分泌，减少 IL-18 和 IL-1β 细胞因子的产生，并下调参与 NLRP3 介导的 RAW264.7 巨噬细胞焦亡的关键蛋白的表达。 BTQ 在 CIA 中的治疗潜力在于其抑制 NLRP3 介导的巨噬细胞焦亡的能力，从而为治疗 RA 提出了一种有前景的策略。© 2024 Wu 等人。

Rheumatoid arthritis (RA) is an autoimmune disease characterized by synovitis and inflammatory cell infiltration. The traditional Chinese medicine prescription, Bitongqing (BTQ) exhibited significant efficacy in the clinical treatment of RA. However, the potential therapeutic mechanisms of BTQ in treating RA have not been fully investigated. This study aims to elucidate the effect of BTQ on collagen-induced arthritis (CIA) rat macrophage pyroptosis, providing a theoretical basis for treating RA.This research employed liquid chromatography-mass spectrometry (LC-MS) to identify the primary components of BTQ. The therapeutic effects of BTQ were evaluated in a rat model of CIA. In vivo experiments were conducted using pathohistological staining, immunofluorescence, micro-CT, and Western blotting. Next, Mouse leukemia cells of monocyte macrophage cells (RAW264.7) were induced to undergo pyroptosis using lipopolysaccharide (LPS) and adenosine triphosphate (ATP), and the impact of BTQ on RAW264.7 macrophages was assessed through cell viability, immunofluorescence analysis, lactate dehydrogenase (LDH) secretion measurement, and Western blotting.BTQ had a therapeutic effect on CIA rats, which was mainly manifested as a reduction in joint inflammation, foot swelling, bone erosion, and amelioration of pathological changes in these rats. Further studies revealed that BTQ inhibited the levels of cytokine production interleukin-18 (IL-18) and interleukin-1β (IL-1β), and likewise, it inhibited the expression of key proteins in the NOD-like receptor thermal protein domain associated protein 3 (NLRP3) mediated pyroptosis in the synovial tissues of CIA rats. The results of in vitro experiments demonstrated that BTQ attenuated LDH secretion, decreased IL-18 and IL-1β cytokine production, and downregulated expression of key proteins involved in the NLRP3-mediated pyroptosis on RAW264.7 macrophages.The therapeutic potential of BTQ in CIA lies in its ability to inhibit NLRP3-mediated macrophage pyroptosis, thereby suggesting a promising strategy for the treatment of RA.© 2024 Wu et al.