Development of the novel therapeutic method for rheumatoid arthritis based on the redox regulation

1.硫氧还蛋白（TRX）转基因小鼠（TRX- tg）的抗关节炎作用由于TRX- tg具有C57BL/6基因背景，且对胶原诱导的关节炎具有抵抗性，因此通过腹腔注射抗胶原II型单克隆抗体鸡尾酒诱导关节炎。(1)野生型小鼠（Wt）在第6天100%诱导关节炎，而TRX-Tg仅在第7天发生关节炎，因此观察到关节炎的晚发性。(2) Wt上，关节红肿明显，关节评分7 ~ 14分（平均10分）。在TRX-Tg中，仅观察到轻微的关节红肿，关节评分为0（无关节炎）至2。(3)组织学上，TRX-Tg与Wt相比，滑膜增生、中性粒细胞浸润和软骨破坏较轻，证实了对软骨凋亡的抑制作用。外源性TRXArthritis的抗关节炎作用以同样的方式在Wt上诱导，每天腹腔注射重组TRX （rTRX）。服用rTRX后，关节炎的发病时间也同样延迟了一天。rTRX对关节肿胀也有抑制作用，但效果不如TRX-Tg。这些结果表明，足够量的TRX对氧化应激的抑制可能导致类风湿关节炎（RA）的新治疗方法。腺苷对滑膜细胞TRX表达的影响为探讨甲氨蝶呤抗风湿作用的效应分子腺苷的抗氧化功能是否与TRX有关，我们用腺苷孵育RA患者滑膜细胞，采用ELISA和流式细胞术检测TRX表达的变化。腺苷对TRX水平没有影响，这表明腺苷的抗风湿作用可能至少与TRX没有直接关系。

1.Anti-arthritis effects of thioredoxin (TRX) transgenic mice (TRX-Tg)Since TRX-Tg have C57BL/6 background and are resistant to collagen-induced arthritis, arthritis was induced by the intraperitoneal injection of anti-collagen type II monoclonal antibody cocktail.(1)Arthritis was 100% induced at day 6 on wild-type mice (Wt) while only few TRX-Tg developed arthritis on day 7, thus the late onset of arthritis was observed.(2)On Wt, joint redness and swelling were significant and the joint scores were 7 to 14 (average 10). On TRX-Tg only slight, if any, joint redness and swelling were observed and the joint scores were 0 (no arthritis) to 2.(3)Histologically synovial proliferation, neutrophil infiltration and cartilage destruction were mild in TRX-Tg compared to Wt. Inhibition of cartilage apoptosis was proved.2.Anti-arthritis effects of exogenous TRXArthritis was induced in the same manner on Wt and recombinant TRX (rTRX) was injected intraperitoneally every day. Onset of arthritis was similarly delayed one day by rTRX administration. Inhibition of joint swelling was also achieved by rTRX administration but not as strong as was observed on TRX-Tg.These results indicate that the inhibition of oxidative stress by enough amount of TRX may lead to the novel therapeutic method for rheumatoid arthritis (RA).3.The effect of adenosine on TRX expression of synoviocytesTo investigate if the anti-oxidative function of adenosine, the effector molecule of anti-rheumatic effects of methotrexate, is related to the TRX, synoviocytes from RA patients were incubated with adenosine and the change of TRX expression was estimated by ELISA and flow cytometry. Adenosine had no influence to the TRX levels, which suggests that anti-rheumatic effects of adenosine may not be at least directly related to the TRX.