Pan Zhenzhen, Li Fangchan, Xu Yujie, Ye Huimin, Liu Jiahui, Wang Zhenhua, Deng Changsheng, Song Jianping, Mei Manxue, Li Changqing
Artemisinin Research Center, Guangzhou University of Chinese Medicine, Guangzhou, China.
Experimental Teaching Center, Guangxi University of Chinese Medicine, Nanning, China.
Front Pharmacol. 2024 Jul 23;15:1423884. doi: 10.3389/fphar.2024.1423884. eCollection 2024.
Fu-zi decoction (FZD) has a long history of application for treating Rheumatoid arthritis (RA) as a classic formulation. However, its underlying mechanisms have not been fully elucidated. This study aimed to decipher the potential mechanism of FZD in treating RA, with a specific focus on receptor activator of nuclear factor κB/receptor activator of nuclear factor κB ligand (RANK/RANKL) signaling pathway.
The impact of FZD on RA was investigated in collagen-induced arthritis rats (CIA), and the underlying mechanism was investigated in an osteoclast differentiation cell model. , the antiarthritic effect of FZD at various doses (2.3, 4.6, 9.2 g/kg/day) was evaluated by arthritis index score, paw volume, toe thickness and histopathological examination of inflamed joints. Additionally, the ankle joint tissues were determined with micro-CT and safranin O fast green staining to evaluate synovial hyperplasia and articular cartilage damage. , osteoclast differentiation and maturation were evaluated by TRAP staining in RANKL-induced bone marrow mononuclear cells. The levels of pro- and anti-inflammatory cytokines as well as RANKL and OPG were evaluated by ELISA kits. In addition, Western blotting was used to investigate the effect of FZD on RANK/RANKL pathway activation both and .
FZD significantly diminished the arthritis index score, paw volume, toe thickness and weigh loss in CIA rats, alleviated the pathological joint alterations. Consistent with results, FZD markedly inhibited RANKL-induced osteoclast differentiation by decreasing osteoclast numbers in a dose-dependent manner. Moreover, FZD decreased the levels of pro-inflammatory cytokines IL-6, IL-1β and TNF-α, while increasing anti-inflammatory cytokine IL-10 level both in serum and culture supernatants. Treatment with FZD significantly reduced serum RANKL levels, increased OPG levels, and decreased the RANKL/OPG ratio. In both and settings, FZD downregulated the protein expressions of RANK, RANKL, and c-Fos, while elevating OPG levels, further decreasing the RANKL/OPG ratio.
In conclusion, FZD exerts a therapeutic effect in CIA rats by inhibiting RANK/RANKL-mediated osteoclast differentiation, which suggested that FZD is a promising treatment for RA.
附子汤（FZD）作为一种经典方剂，在治疗类风湿关节炎（RA）方面有着悠久的应用历史。然而，其潜在机制尚未完全阐明。本研究旨在揭示FZD治疗RA的潜在机制，特别关注核因子κB受体激活剂/核因子κB配体受体激活剂（RANK/RANKL）信号通路。
在胶原诱导性关节炎大鼠（CIA）中研究FZD对RA的影响，并在破骨细胞分化细胞模型中研究其潜在机制。通过关节炎指数评分、 paw体积、趾厚度和炎症关节的组织病理学检查评估FZD在不同剂量（2.3、4.6、9.2 g/kg/天）下的抗关节炎作用。此外，用显微CT和番红O固绿染色测定踝关节组织，以评估滑膜增生和关节软骨损伤。通过抗酒石酸酸性磷酸酶（TRAP）染色评估RANKL诱导的骨髓单核细胞中的破骨细胞分化和成熟。用酶联免疫吸附测定试剂盒评估促炎和抗炎细胞因子以及RANKL和骨保护素（OPG）的水平。此外，用蛋白质免疫印迹法研究FZD在体内和体外对RANK/RANKL途径激活的影响。
FZD显著降低CIA大鼠的关节炎指数评分、 paw体积、趾厚度和体重减轻，减轻关节病理改变。与体内结果一致，FZD通过以剂量依赖性方式减少破骨细胞数量，显著抑制RANKL诱导的破骨细胞分化。此外，FZD降低血清和培养上清液中促炎细胞因子白细胞介素-6（IL-6）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的水平，同时增加抗炎细胞因子IL-10的水平。FZD治疗显著降低血清RANKL水平，增加OPG水平，并降低RANKL/OPG比值。在体内和体外环境中，FZD均下调RANK、RANKL和c-Fos的蛋白表达，同时提高OPG水平，进一步降低RANKL/OPG比值。
总之，FZD通过抑制RANK/RANKL介导的破骨细胞分化在CIA大鼠中发挥治疗作用，这表明FZD是一种有前途的RA治疗药物。
