乌梢蛇Ⅱ型胶原提取及其诱导大鼠关节炎特性的研究  

Extraction of typeⅡcollagen from Zaocys dhumnades (Cantor) and its property for the development of collagen-induced arthritis

作　　者：沈杰 鲍建芳[2] 何东仪 张之澧 倪立青 

机构地区：[1]上海市长宁区光华中西医结合医院风湿科,上海200052 [2]浙江大学医学院细胞与分子生物学实验中心,杭州310006

出　　处：《现代免疫学》2007年第3期223-227,共5页Current Immunology

摘　　要：提取和纯化乌梢蛇Ⅱ型胶原,观察乌梢蛇Ⅱ型胶原诱导大鼠关节炎的特性,为研究乌梢蛇治疗类风湿关节炎的机制提供研究方向。离心提取乌梢蛇Ⅱ型胶原,经DEAE-52纤维素交换柱纯化,斑点免疫渗滤试验和免疫印迹试验进行成分鉴定。乌梢蛇Ⅱ型胶原不完全佐剂免疫大鼠,观察大鼠关节炎发生情况、抗Ⅱ型胶原抗体、CD 4/CD8亚群、血清中TNF-α、IL-10、IL-1β、IL-4的变化。乌梢蛇Ⅱ型胶原提纯品在SDS-PAGE凝胶电泳上呈一条区带,斑点免疫渗滤试验和免疫印迹试验为阳性反应。诱导大鼠关节炎发生率为86.67%,关节炎大鼠血清中抗Ⅱ型胶原抗体明显增高(P<0.01),CD4 T细胞亚群和CD4^+/CD8^+增高(P<0.05),TNF-α含量增加(P<0.01),IL-10含量降低(P<0.01),血清中IL-1β和IL-4含量无变化。结果显示,乌梢蛇Ⅱ型胶原能通过免疫方法可以诱导大鼠产生多关节炎,并且体内有自身免疫反应的表现。The type Ⅱ collagen （CII） was extracted from Zaocys dhumnades （Caantor） and its property to develop the collagen-induced arthritis in rats was observed in order to explore the possibility to use Z. dhurnnades for the treatment of rheuma toid arthritis. CII was isolated from Z. dhurnnades by centrifugation and then purified through DEAE-52 anion exchange column and identified by dot-immunofiltration and immunoblotting assay with monoclonal antibody against cattle type Ⅱ collagen. The purified CII of Z. dhurnnades with incomplete adjuvant was then injected to the back of rats,and the development of arthritis in rats was observed. The antibodies agaist CII ratio of CD4/CD8 subsets and changes in the production of cytokines, such as TNF-α,IL-10,IL-β and IL-4 in serum of rats were determined. It was found that the purified CII of Z. dhunades （Cantor） showed a single band on the SDS-PAGE gel electrophoresis and it was confirmed by dot immunofiltration and immunoblotting assays. The incidence of development of arthritis in rats injected with CUU of Z. dhurnnades was 86.67%. In rats with arthri tis, the ratio of CD4/CD8 subset and serum level of antibodies against CII and TNF-a were increased, but that of IL-10 was decreased. Meanwhile, the serum levels of 1L-113 and IL-4 demonstrated no significant difference. These results suggest that type II collagen of Z. dhumnades （Cantor） can induce arthritis in rats with appearance of autoimmune reactions in vivo.

关 键 词：Ⅱ型胶原 乌梢蛇 类风湿关节炎 

分 类 号：R392.12[医药卫生—免疫学]