正清风痛宁联合甲氨蝶呤对CIA大鼠血清RANKL、MMPs表达的影响

摘要

目的 通过观察正清风痛宁联合甲氨蝶呤(MTX)对胶原诱导的关节炎大鼠模型血清核刺激因子-κB受体配体(RANKL)和金属蛋白酶(MMPs)的表达影响,探讨其防治类风湿性关节炎(RA)骨质破坏的作用机制.方法 建立Ⅱ型胶原蛋白诱导的SD大鼠关节炎模型.将30只SD大鼠随机分为4组:正常组、模型组、MTX组和联合组(正清风痛宁+MTX).采用关节评分法评估关节炎症程度,并用ELISA法分析大鼠血清中OPG、RANKL、MMP-3、MMP-13的表达水平.结果 ①SD大鼠造模后,炎症迅速发展,关节肿胀在第21天最为明显,然后逐渐消退,在第35天左右会出现第2个炎症高峰.MTX组和联合组均可减轻关节肿胀,关节炎指数评分在第46天与造模组比较差异有统计学意义(P＜0.05)；②与模型组比较,MTX组、联合组均提高OPG的表达水平,升高OPG与RANKL的比值；联合组可降低血清RANKL的表达水平,差异有统计学意义(P＜0.05)；联合组与MTX组比较,血清RANKL的表达水平下降更为明显,OPG与RANKL的比值也有显著上升,差异有统计学意义(P＜0.05)；③MTX组和联合组都可明显抑制血清MMP-3和MMP-13的表达水平,同时联合组血清MMP-3的表达水平低于MTX组,差异有统计学意义(P＜0.05).结论 正清风痛宁联合MTX能够较好地控制炎症,并可协同延缓骨破坏的发生,可能与其通过对MMP-3、MMP-13的抑制,提高OPG/RANKL的比例有关.%OBJECTIVE Through the observation of effects of Zhengqing Fengtongning combined with methotrexate on the expression of RANKL and MMPs in the serum of collagen induced arthrites rats, the action mechanism of it on the prevention of RA bone destruction is discussed. METHODS SD arthritis rats model induced by CollagenII was made and 30 SD rats, were randomized into 4 groups: normal group, model group, MTX group and joint group (Zhengqing Fengtongning +MTX). The severity of articular inflammation was assessed by joint scoring method and the expression level of OPG, RANKL、 MP-3 and MMP-13 in serum was analyzed by the method of ELISA. RESULTS ①After being made model, the inflammation developed rapidly and appeared most obvious on the 21st day, and then gradually subsided. The second summit of inflammation appeared about the 35th day. MTX group and Joint group can reduce the articular swelling in rats. There was significant difference arthritis indexs between these two groups and the immodel group on the 46th day (P<0. 05). ② Compared with the model group, MTX group and joint group can enhance the expression level of OPG and increase the tatio of OPG and RANKL; joint group can reduce the expression levels of RANKL in serum. The difference had statistical significance (P<0.05). ③The expression levels of MMP-3 and MMP-13 can be significantly prohibited in the joint group and MTX group. And the expression level of MMP-3 in joint group was lower than that in MTX group .The difference was statistically significant (F<0.05). CONCLUSION Zhengqing Fengtongning and MTX can not only control the articular swelling well, but synergistically delay theoccurrence of bone destruction, which maybe related to the inhibition of MMP-3 and MMP-13 and the increase of OPG/RANKL ratio to some extent.