Liu Jing, Liu Ge, Chu Teng, Wu Yue, Yan Xingyu, Pang Liping, Fang Weirong
School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu, People's Republic of China.
Department of Medicine, Ningbo Liwah Pharmaceutical Co., Ltd, Ningbo, Zhejiang, People's Republic of China.
J Inflamm Res. 2025 Jul 29;18:10147-10161. doi: 10.2147/JIR.S527354. eCollection 2025.
The cascade of inflammation caused by the activation of NLRP3 inflammasome plays an important role in monosodium urate (MSU)-induced acute gouty arthritis (AGA) in mice. This study aimed to evaluate the therapeutic effect of total glucosides of paeony (TGP) in AGA mice and elucidate the underlying anti-inflammatory mechanism.
MSU crystals were injected into the ankle joint to establish the AGA model in mice, and bone marrow-derived macrophages (BMDMs) were primary cultured and stimulated with MSU and LPS to model inflammatory conditions in vitro. Ankle diameter was measured to quantify joint swelling. Pain was assessed by the fifty percent paw withdrawal threshold (PWT) test and the bipedal support test. Pro-inflammatory cytokine levels and arthropathological damage were analyzed by ELISA, H&E staining, and immunohistochemistry staining. The mRNA and protein expression of NLRP3 inflammasome-associated signaling pathways were determined by RT-qPCR and Western blotting, respectively. Mitochondrial oxidative damage was evaluated using flow cytometry and immunofluorescence staining.
Four-week TGP administration with 360, 540, 720 mg/kg respectively significantly attenuated joint swelling and pain (P < 0.05, P< 0.01), reduced pro-inflammatory cytokine secretion (P < 0.01), and ameliorated joint pathology and macrophage infiltration (P < 0.01) in AGA mice. In BMDMs, TGP (30, 60 μg/mL) treatment notably suppressed mRNA and protein expression of NLRP3, caspase-1, and IL-1β, and lowered inflammatory factor secretion (P < 0.05, P< 0.01). Additionally, cellular and mitochondrial ROS levels were both significantly decreased (P < 0.01), while ATP concentration markedly increased (P < 0.01), suggesting improved mitochondrial function.
TGP significantly alleviated experimental AGA by reducing macrophage infiltration and suppressing the NLRP3-mediated inflammatory response.
NLRP3炎性小体激活所引发的炎症级联反应在小鼠尿酸钠（MSU）诱导的急性痛风性关节炎（AGA）中起重要作用。本研究旨在评估白芍总苷（TGP）对AGA小鼠的治疗效果，并阐明其潜在的抗炎机制。
将MSU晶体注射到小鼠踝关节以建立AGA模型，原代培养骨髓来源的巨噬细胞（BMDM），并用MSU和LPS刺激以在体外模拟炎症状态。测量踝关节直径以量化关节肿胀。通过50%足趾撤离阈值（PWT）试验和双足支撑试验评估疼痛。通过ELISA、苏木精-伊红（H&E）染色和免疫组织化学染色分析促炎细胞因子水平和关节病理损伤。分别通过逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法测定NLRP3炎性小体相关信号通路的mRNA和蛋白质表达。使用流式细胞术和免疫荧光染色评估线粒体氧化损伤。
分别以360、540、720 mg/kg的剂量连续四周给予TGP可显著减轻AGA小鼠的关节肿胀和疼痛（P < 0.05，P < 0.01），减少促炎细胞因子分泌（P < 0.01），并改善关节病理和巨噬细胞浸润（P < 0.01）。在BMDM中, TGP（30、60 μg/mL）处理显著抑制NLRP3、半胱天冬酶-1（caspase-1）和白细胞介素-1β（IL-1β）的mRNA和蛋白质表达，并降低炎症因子分泌（P < 0.05，P < 0.01）。此外，细胞和线粒体活性氧水平均显著降低（P < 0.01），而三磷酸腺苷（ATP）浓度显著升高（P < 0.01），提示线粒体功能改善。
TGP通过减少巨噬细胞浸润和抑制NLRP3介导的炎症反应，显著减轻实验性AGA。
