构建双基因共表达腺病毒载体转染关节炎模型大鼠的意义  

Transfection of double gene co-expressing adenovirus vector into arthritis rats

作　　者：樊萍[1] 何岚[1] 郝志明[1] 蒲丹[1] 吕晓虹[1] 孙怡宁[1] 胡楠[1] 王研华 丁小明[2] 李杨[2] 薛武军[2] 

机构地区：[1]西安交通大学医学院第一附属医院风湿免疫科,陕西省西安市710061 [2]西安交通大学医学院第一附属医院肾移植科,陕西省西安市710061

出　　处：《中国组织工程研究》2015年第18期2825-2830,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目资助(81400677);陕西省科学技术研究发展计划项目资助(2012K16-09-02)~~

摘　　要：背景:淋巴细胞异常激活以及核因子κB依赖非特异性的炎症反应是类风湿关节炎关节组织炎症损害的两个重要方面。共刺激信号CD40/CD40L是T细胞识别、活化中最重要的共刺激分子。IκBα有效的抑制核因子κB的途径,可以在中心环节上抑制炎症反应的产生,抑制炎症因子滑膜组织的损害。目的:采用CD40LIg-IRES2-IκBα双基因表达的腺病毒载体转染到关节炎动物模型,探索双基因共表达腺病毒载体转染对免疫性关节炎的治疗效果。方法:构建pA dC D40LIg-IRES2-IκBα双基因共表达腺病毒载体。采用含1 g/LⅡ型胶原蛋的完全弗氏佐剂皮下多点注射构建关节炎Wistar大鼠模型。将20只构建成功的关节炎大鼠模型分为未处理组和转染组,分别给予2组大鼠四肢末端关节腔注射生理盐水和pA d CD40LIg-IRES2-IκBα腺病毒载体。结果与结论:转染14 d后,与未处理组相比,转染组大鼠关节炎评分、关节液中淋巴细胞CD40L表达水平、关节滑膜组织中核因子κB p65表达水平、关节液内白细胞介素2、白细胞介素6、肿瘤坏死因子α、间质金属蛋白酶3和间质金属蛋白酶9的水平降低。提示共表达CD40LIg和IκBα腺病毒载体局部转染可有效抑制关节炎大鼠关节炎症状,降低关节腔内炎性细胞因子及炎性分子在滑膜组织中的表达,取得良好的治疗效果。BACKGROUND：Abnormal activation of lymphocytes and nuclear factorκB-dependent non-specific inflammation are two major manifestations of joint damage in rheumatoid arthritis. Co-stimulatory signal CD40/CD40L is the dominant co-stimulatory factor in the recognition and activation of T cel s. IκBαeffectively inhibits nuclear factorκB pathway, prevent the inflammation in the central link, and suppress the damage caused by inflammatory factor in the synovial tissue. OBJECTIVE：To investigate the therapeutic effect of double gene co-expressing adenovirus vector on arthritis based on an arthritis model rat transfected by CD40LIg-IRES2-IκBαco-expressing adenovirus vector. METHODS：The pAdCD40LIg-IRES2-IκBαco-expressing adenovirus vector was established. Arthritic model was established through multi-subcutaneous injections of complete Freund＇s adjuvant of type col agen II （1 g/L） into Wistar rats. Then 20 arthritic rats were divided into two groups：untreated group and transfection group, receiving an injection of saline and pAdCD40LIg-IRES2-IκBαadenovirus vector to distal joint cavity of limbs, respectively. RESULTS AND CONCLUSION：At 14 days post-transfection, compared with the untreated group, the mean arthritis index score, the CD40L expression of lymphocytes in synovial fluid, the nuclear factor-κB p65 expression in synovial tissue, and levels of interleukin-2, interleukin-6, tumor necrosis factor-α, matrix metal oproteinase-3 and matrix metal oproteinase-9 in synovial fluid of rats in transfection group were significantly lower than those in＆nbsp;untreated group. Focal transfection of the CD40LIg-IκBαco-expression adenovirus vector can effectively inhibit arthritic symptoms, and reduce the expressions of inflammatory cytokine in synovial fluid and inflammatory molecule in synovial tissue of arthritic rats, which shows good therapeutic effect.

关 键 词：实验动物 骨及关节损伤模型 关节炎 类风湿 CD40L IΚBΑ 核因子κB 共刺激信号 炎症 滑膜 国家自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]