周期联合甲氨蝶呤和环磷酰胺对大鼠胶原诱导性关节炎滑膜细胞周期蛋白D1表达的研究  

Effect of combined methotrexate and cyclophosphamide therapy on the cyclin D1 expression in the synovium of rats with collagen induced arthritis

作　　者：王彩虹[1] 李小峰[1] 张奉春[2] 曲世晶[1] 梁美娥[1] 

机构地区：[1]山西医科大学第二医院风湿免疫科,太原030001 [2]中国医学科学院 中国协和医科大学 北京协和医院风湿免疫科

出　　处：《中华风湿病学杂志》2008年第12期812-815,I0001,共5页Chinese Journal of Rheumatology

基　　金：基金项目：山西省自然科学基金资助项目（2007011116）;山西省卫生厅科技攻关项目（200614）;山西医科大学第二医院博士启动基金资助项目（20070405）;山西医科大学第二医院青年科学基金资助项目（20060101）

摘　　要：目的通过研究大鼠胶原诱导性关节炎（CIA）滑膜组织细胞周期蛋白D1（cvclin D1）的表达以及甲氨蝶呤（MTX）和环磷酰胺（CTX）对其的影响，从周期调控的角度探讨两者联合治疗类风湿关节炎（RA）可能的协同效应机制。方法建立Ⅱ型胶原诱导的雌性Wistar大鼠CIA模型，随机分为正常对照组、CIA模型对照组、小剂量MTX治疗组（每周0．9mg／kg）、大剂量MTX治疗组（每周2．7mg／kg）、小剂量CTX间歇治疗组（每3周24mg／kg）、联合治疗组（MTX每周0．9mg／kg＋CTX每3周24mg／kg）。治疗24周后全部动物处死取材，再经固定、脱钙、包埋，原位杂交法检测滑膜细胞周期蛋白cyclin D1 mRNA。结果CIA模型组大鼠关节滑膜组织的cyclin D1 mRNA的阳性染色强度明显高于正常对照组（P〈0．05），各治疗组滑膜cyclin D1 mRNA下调，联合治疗组cvclin D1 mRNA表达最低，各组间平均灰度值差异有统计学意义（P〈0．05）。MTX与CTX存在药物交互作用（P〈0．05），联合治疗组优于单用药治疗组。结论提示MTX和CTX联合治疗为协同效应，可能的重要机制是使滑膜cvclin D1 mRNA下调，从而抑制了滑膜细胞异常增殖。Objective To explore effects and possible mechanisms of combination therapy with methotrexate （MTX） and low dose eyclophosphamide （CTX） intermittently on the cyclin D 1 expression in the synovium of collagen induced arthritis rats. The outcome is analyzed. Methods After CIA experimental models were successfully established in female Wistar rats, they were randomly divided into CIA model group, low-dose MTX treatment group （0.9 mg· kg^-1·W^-1）, high-dose MTX treatment group （2.7 mg· kg^-1·W^-1, lowdose CTX treatment group [ 24 mg·kg^-1·（3W）^-1 ], and low-dose MTX ＋ low-dose CTX treatment group （MTX 0.9 mg ·kg^-1 ·W^-1, CTX [24 mg ·kg^-1·（3W）^-1]. All of the rats were sacrificed 24 weeks after the first immunization, the specimens of ankle joints were exposed, fixed, decalcified, wrapped and cut into slices. The Cyclin D1 mRNA levels were examined by situ hybridization. Results The mRNA level of cyelin D1 in CIA synovial lining layer was significantly higher than that of normal controls, which suggested that in situ proliferation of synovioeytes might contribute to the thickening of rheumatoid synovial lining layer. The synovial expression intensity of the mRNA level of cyclin D1 in the treatment groups was decreased evidently. The mRNA level of eyclin D1 of the combination treatment group was higher than other groups, which was statistically significant （P〈0.05）.Conclusion This is the first research on the effects of MTX combined with CTX intermittently based on cell cycles in CIA animal model. Combination therapy seems to be more effective than single-agent therapy. The results suggest that combining cell cycle stage specific agent MTX with cell cycle stage nonspecifie agent CTX may he synergistic in the treatment of RA. Inhibition of the increased expression of the synovial eyclin D1 mRNA maybe an important mechanism of the MTX and CTX combination treatment for RA.

关 键 词：关节炎 实验性 细胞周期蛋白D1 甲氨蝶呤 环磷酰胺 

分 类 号：R686[医药卫生—骨科学]