抗骨桥蛋白抗体对胶原诱导大鼠类风湿关节炎作用的研究

Study on the Effect of Anti Osteopontin Antibody in Treating Rats' Collagen Induced Rheumatoid Arthritis

目的研究抗骨桥蛋白抗体对类风湿关节炎大鼠滑膜组织的保护作用及机制。方法 40只SD大鼠,随机抽取6只为正常对照组,34只建立胶原诱导的类风湿关节炎模型;选取24只关节炎指数>2的大鼠(其余10只剔除),随机分为模型组、抗骨桥蛋白抗体低、中和高剂量治疗组。记录大鼠体质量、关节炎指数和左后足足趾容积。第35天处死大鼠,取其关节进行HE染色,观察关节病理学改变;Western blot检测滑膜组织内骨桥蛋白(OPN)和肿瘤坏死因子α(TNF-α)蛋白表达情况;ELISA检测大鼠血液内OPN和TNF-α含量的变化。结果治疗组大鼠体质量第16天平稳上升,给药后关节肿胀逐步缓解。与正常组比较,模型组体质量、足趾容积和关节炎指数差异有统计学意义(P<0.05)。与模型组比较,治疗组体质量、足趾容积和关节炎指数差异有统计学意义(P<0.05)。治疗组可见滑膜细胞增生明显减轻,少量炎性细胞浸润,关节软骨表面光滑,未见破坏。与正常组比较,模型组OPN蛋白表达明显升高,差异有统计学意义(P<0.05);与模型组比较,治疗组OPN蛋白表达明显降低,差异有统计学意义(P<0.05);与正常组比较,模型组TNF-α表达明显升高,差异有统计学意义(P<0.05)。结论抗骨桥蛋白抗体可明显减轻CIA模型大鼠的关节症状,抑制滑膜细胞增生,减轻炎性反应。抗骨桥蛋白抗体保护CIA模型大鼠关节过程中,抑制大鼠滑膜组织及血清中OPN的表达,不影响TNF-α的表达。

Objective To investigate the anti-OPN antibody′s protective effect towards the synovial membrane of rats with rheumatoid arthritis,in-cluding the mechanism of such protective action. Methods From a total of 40 SD rats,6 were randomly selected as the normal control group while the remaining 34 were used to establish the collagen induced arthritis model. A total of 24 rats with arthritis index higher than 2 were chosen（exclud-ing the rest 10 rats）,and randomly divided into model group and groups being treated with low,medium and high dose of anti-OPN anti-body,re-spectively. Weight,arthritis index and the volume of the left hind leg toe of the rats were recorded. Then on day 35,the rats were killed and their joints were taken out to make HE colored sections for observation of the pathological changes. The synovial membrane of each rat was taken out and WB method was used to test the protein expression of OPN and TNF-αin such membrane. ELISA method was used to test the change of the content of OPN and TNF-αin each rat′s blood. Results The rats in treatment group gained weight steadily from day 16 and their swollen joints were gradu-ally relieved upon medication. Comparing with the control group,the differences in weight,toe volume and arthritis index of the rats in model group were statistically significant（P〈0.05）. Comparing with the model group,the differences in weight,toe volume and arthritis index of the rats in the treatment groups were also statistically significant（P〈0.05）. Pathologic change of synovial membrane：for rats in treatment groups,synovial cell hy-perplasia was visibly reduced with a little inflammatory cell infiltration and smooth articular cartilage surfaces with no visible damage. Expression of OPN and TNF-αin synovial membrane and blood serum of rats：Judging from the OPN measured in synovial membrane and blood serum,we could see that the expression of OPN in rats of the model group increased obviously if compared with the control group. Comparing with the model group, the expression of OPN in the treatment groups was reduced（P〈0.05）. Judging from the TNF-αmeasured in synovial membrane and blood serum, we could see that the expression of TNF-αin rats of the model group increased obviously if compared with the control group,thus such difference was statistically significant between different groups（P〈0.05）. Conclusion The anti-OPN anti-body could significantly reduce the articular symp-toms of the rats with collagen induced arthritis,restrain the synovial cell hyperplasia and alleviate the inflammatory reaction. In the process of protect-ing the joints of the rats with collagen induced arthritis,anti-OPN anti-body restrained only the expression of OPN in the synovial membrane and blood serum,leaving the expression of TNF-αunaffected.