目的 研究全蝎-土鳖虫药对减轻胶原诱导的关节炎（collagen-induced arthritis，CIA）模型小鼠关节骨破坏的作用以及对破骨细胞分化的核因子-κB（nuclear factor-κB，NF-κB）信号通路的影响，探讨全蝎-土鳖虫药对治疗类风湿关节炎的机制。方法 DBA/1J小鼠随机分为正常组、模型组及全蝎-土鳖虫低、高剂量组和甲氨蝶呤组。除正常组外，其余组建立CIA模型，正常组和模型组ig纯净水，其余各组给药干预4周。测定各组小鼠足趾肿胀情况；采用Micro-CT扫描的方法重建小鼠关节3D图并分析相关骨参数；苏木素-伊红（HE）染色观察小鼠踝关节病理变化；抗酒石酸酸性磷酸酶（tartrate-resistant acid phosphatase，TRAP）染色检测关节中破骨细胞分化情况。体外实验考察全蝎-土鳖虫药对对核因子-κB受体活化因子配体（receptor activator of nuclear factor-κB ligand，RANKL）处理的小鼠骨髓来源的巨噬细胞诱导破骨细胞的影响；免疫荧光染色检测NF-κB p65核定位情况。Western blotting检测破骨细胞分化、CIA小鼠骨组织中NF-κB信号通路相关蛋白表达。结果 各给药组显著降低CIA模型小鼠足趾肿胀（P＜0.05、0.01）；与模型组比较，Micro-CT 3D重建结果表明，全蝎-土鳖虫药对明显抑制CIA模型小鼠骨破坏，保留小鼠骨结构完整性，显著改善骨参数（P＜0.05、0.01）；全蝎-土鳖虫药对显著减少关节TRAP染色阳性细胞数量（P＜0.01）；全蝎-土鳖虫药对高剂量组显著降低CIA小鼠骨组织中NF-κB p65磷酸化水平（P＜0.01）。体外实验结果显示，全蝎-土鳖虫药对显著抑制破骨细胞的分化（P＜0.01），抑制NF-κB p65核定位；与RANKL对照组比较，全蝎-土鳖虫药对组p65、核因子-κBα抑制蛋白（inhibitor of nuclear factor-κB alpha，IκBα）磷酸化水平明显降低（P＜0.05、0.01）。结论 全蝎-土鳖虫药对能够减轻CIA小鼠骨破坏，可能是通过抑制NF-κB信号通路活化、降低破骨细胞的分化而发挥作用。

Objective To investigate the effect of Quanxie (Scorpio)-Tubiechong (Eupolyphaga Steleophaga) drug pair on alleviating joint bone destruction in collagen-induced arthritis (CIA) mice and nuclear factor-κB (NF-κB) signaling pathway for osteoclast differentiation, and explore the mechanism therapy for treating rheumatoid arthritis. Methods DBA/1J mice were randomly divided into normal group, model group, Scorpio-Eupolyphaga Steleophaga low-and high-dose groups, and methotrexate group. Except for the normal group, the other groups were established with CIA models. The normal group and model group were treated with purified water, the other groups were treated with medication for four weeks. The swelling of toes of mice in each group was measured; Micro CT scanning method was used to reconstruct 3D images of mouse joints and analyze relevant bone parameters; HE staining was used to observe the pathological changes of the mouse ankle joint; Tartaric acid resistant acid phosphatase (TRAP) staining was used to detect the differentiation of osteoclast in joints. In vitro experimental study on the effect of Scorpio-Eupolyphaga Steleophaga drug pair on receptor activator of nuclear factor-κB ligand (RANKL) treated mice bone marrow derived macrophages induced osteoclast. Immunofluorescence staining was used to detect NF-κB p65 nuclear localization. Western blotting was used to detect the differentiation of osteoclast and NF-κB signaling pathway related protein expressions in bone tissue of CIA mice. Results Each administration group significantly reduced toe swelling in CIA model mice (P < 0.05, 0.01); Compared with model group, Micro-CT 3D reconstruction results showed that Scorpio-Eupolyphaga Steleophaga drug pair significantly inhibited bone destruction in CIA model mice, preserved bone structural integrity of mice, and significantly improved bone parameters (P < 0.05, 0.01); Scorpio-Eupolyphaga Steleophaga drug pair significantly reduced the number of TRAP positive cells in joints (P < 0.01); Scorpio-Eupolyphaga Steleophaga drug pair high-dose group significantly decreased NF-κB p65 phosphorylation level in bone tissue of CIA mice (P < 0.01). The results of in vitro experiment showed that Scorpio-Eupolyphaga Steleophaga drug pair significantly inhibited the differentiation of osteoclast and NF-κB p65 nuclear localization; Compared with RANKL control group, p65 and inhibitor of nuclear factor-κB alpha (IκBα) phosphorylation levels were significantly decreased in Scorpio-Eupolyphaga Steleophaga drug pair group (P < 0.05, 0.01). Conclusion Scorpio-Eupolyphaga Steleophaga drug pair can alleviate bone damage in CIA mice, which may be through inhibiting NF-κB signaling pathway activation and reducing the differentiation of osteoclast.

R285.5

江苏省中医药科技发展专项（2020ZX16）；国家自然科学基面上项目（81773973）；南京中医药大学自然科学基金资助项目（XZR2021037）