PDF(Pubmed)
Abstract：
UNASSIGNED: 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.
UNASSIGNED: 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. In vivo, 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. In vitro, 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 in vivo and in vitro.
UNASSIGNED: FZD significantly diminished the arthritis index score, paw volume, toe thickness and weigh loss in CIA rats, alleviated the pathological joint alterations. Consistent with in vivo 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 in vivo and in vitro settings, FZD downregulated the protein expressions of RANK, RANKL, and c-Fos, while elevating OPG levels, further decreasing the RANKL/OPG ratio.
UNASSIGNED: 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.
UNASSIGNED: 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. In vivo, 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. In vitro, 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 in vivo and in vitro.
UNASSIGNED: FZD significantly diminished the arthritis index score, paw volume, toe thickness and weigh loss in CIA rats, alleviated the pathological joint alterations. Consistent with in vivo 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 in vivo and in vitro settings, FZD downregulated the protein expressions of RANK, RANKL, and c-Fos, while elevating OPG levels, further decreasing the RANKL/OPG ratio.
UNASSIGNED: 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.
摘要：
■复子汤（FZD）作为经典制剂在治疗类风湿性关节炎（RA）方面具有悠久的应用历史。然而,其潜在机制尚未完全阐明。本研究旨在破译FZD治疗RA的潜在机制，核因子κB受体活化因子/核因子κB受体活化因子配体(RANK/RANKL)信号通路具有特异性。
■在胶原诱导的关节炎大鼠（CIA）中研究了FZD对RA的影响，并在破骨细胞分化细胞模型中研究了其潜在机制。在体内,通过关节炎指数评分评估FZD在各种剂量（2.3，4.6，9.2g/kg/天）的抗关节炎作用，爪子体积,脚趾厚度和关节发炎的组织病理学检查。此外,采用显微CT和番红花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。体外，通过TRAP染色在RANKL诱导的骨髓单个核细胞中评估破骨细胞的分化和成熟。通过ELISA试剂盒评估促炎和抗炎细胞因子以及RANKL和OPG的水平。此外,Western印迹用于研究FZD在体内和体外对RANK/RANKL途径激活的影响。
■FZD显著降低关节炎指数评分，爪子体积,CIA大鼠的脚趾厚度和体重减轻，减轻病理性关节改变。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而显著抑制RANKL诱导的破骨细胞分化。此外，FZD降低促炎细胞因子IL-6、IL-1β和TNF-α水平,同时增加血清和培养上清液中的抗炎细胞因子IL-10水平。FZD治疗显著降低血清RANKL水平，OPG水平升高,并降低RANKL/OPG比值。在体内和体外环境中，FZD下调RANK的蛋白表达，RANKL,c-Fos,在提高OPG水平的同时,进一步降低RANKL/OPG比值。
■总而言之，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治疗方法。
■在胶原诱导的关节炎大鼠（CIA）中研究了FZD对RA的影响，并在破骨细胞分化细胞模型中研究了其潜在机制。在体内,通过关节炎指数评分评估FZD在各种剂量（2.3，4.6，9.2g/kg/天）的抗关节炎作用，爪子体积,脚趾厚度和关节发炎的组织病理学检查。此外,采用显微CT和番红花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。体外，通过TRAP染色在RANKL诱导的骨髓单个核细胞中评估破骨细胞的分化和成熟。通过ELISA试剂盒评估促炎和抗炎细胞因子以及RANKL和OPG的水平。此外,Western印迹用于研究FZD在体内和体外对RANK/RANKL途径激活的影响。
■FZD显著降低关节炎指数评分，爪子体积,CIA大鼠的脚趾厚度和体重减轻，减轻病理性关节改变。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而显著抑制RANKL诱导的破骨细胞分化。此外，FZD降低促炎细胞因子IL-6、IL-1β和TNF-α水平,同时增加血清和培养上清液中的抗炎细胞因子IL-10水平。FZD治疗显著降低血清RANKL水平，OPG水平升高,并降低RANKL/OPG比值。在体内和体外环境中，FZD下调RANK的蛋白表达，RANKL,c-Fos,在提高OPG水平的同时,进一步降低RANKL/OPG比值。
■总而言之，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治疗方法。
