复方大黄散对类风湿关节炎大鼠MMP-10及IL-33的影响

Effects of the combination of compound rhubarb powder and methotrexate on the expression of MMP-10 and IL-33 in rheumatoid arthritis rats

目的观察中药复方大黄散(CRP)外敷对类风湿关节炎(RA)大鼠的治疗作用及对MMP-10及IL-33的影响,探讨复方大黄散治疗RA的潜在机制。方法将72只大鼠分成正常组、模型组、扶他林组、甲氨蝶呤组、复方大黄散组、联合组(复方大黄散+甲氨蝶呤组),除正常组外,以完全弗氏佐剂(CFA)诱导大鼠RA模型,造模第8天后,各治疗组分别给药。用跖围法测量大鼠足跖肿胀度;治疗6周后,光镜下观察各组大鼠右踝关节滑膜病理学形态变化。Western blot检测大鼠软骨中MMP-10及IL-33的蛋白表达水平;RT-PCR法测定软骨中IL-33、MMP-10的m RNA表达水平。结果模型组大鼠右后足关节红肿明显,伴活动量减少及跛行,软骨中MMP-10及IL-33水平均明显升高(P<0.05),各用药组足跖肿胀较模型组显著减轻(P<0.05),复方大黄散组与扶他林组、甲氨蝶呤组之间无明显差异(P>0.05),联合组大鼠较其它组减轻最明显(P<0.05)。治疗6周后,与模型组比较,各用药组软骨蛋白表达及m RNA的表达含量MMP-10、IL-33水平均显著下降(P<0.05)。光镜下观察各用药组大鼠关节组织中炎症细胞浸润减少,血管翳生成减少,关节腔间隙增宽,关节软骨破坏减轻。结论复方大黄散外敷能明显减轻RA大鼠关节肿胀,作用与扶他林、甲氨蝶呤相当,联合甲氨蝶呤疗效更佳,其机制可能与调控MMP-10、IL-33等细胞因子相关。

This study performed to explore the effects of the compound rhubarb powder（CRP） on the expressions of MMP-10 and IL-33 in rheumatoid arthritis（RA） rats, and then investigate the mechanism of CRP in the treatment of RA. Seventy-two SD rats were randomly divided into six groups： normal, model, voltaren,methotrexate, CRP, CRP plus methotrexate, with 12 rats in each group. The model group was induced by intradermal injection of complete freund′ sadjuvant（CFA）, and the swelling degree of rear foot of rats were measured by plantar circumference. Drugs were administered to each treatment group from the ninth day after modeling.Western blot and quantitative real-time PCR were performed to analyze the expression of MMP-10 and IL-33 in the ankle joint, while the histopathology of ankle joint was observed under light microscopy after the treatment for six weeks. Data revealed that rear foot of the rats were severe swelling, while erythema and the expression of MMP-10 and IL-33 were significantly increased（P〈0.05） in model group with activity decrease and limp; compared with model group, treated groups showed significantly reduction in paw swelling（P〈0.05）, especially CRP plus methotrexate-treated rats（P〈0.05）, but there were no significant difference among CRP-, voltaren- and methotrexate-treated rats（P〉0.05）. Western blot and quantitative real-time PCR results showed that there was a significant decrease in the expressions of MMP-10 and IL-33 in treated groups compared with model group（P〈0.05）. Under light microscopy, histopathology of ankle joints demonstrated that the inflammatory cells infiltration and pannus reduced, the joint cavity gap widen and the bone erosion decreased. In conclusion, CRP can obviously reduce the swelling degree, especially in combination with methotrexate, which is comparable to voltaren- and methotrexate-treatment. The mechanism may involve in regulating the expressions of cytokines such as MMP-10 and IL-33.