【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。更多还原

【Abstract】 The typeⅡcollagen(CII) was extracted from Zaocys dhumnades(Caantor)and its property to develop the colla- gen-induced arthritis in rats was observed in order to explore the possibility to use Z.dhumnades for the treatment of rheuma- toid arthritis.CII was isolated from Z.dhumnades by centrifugation and then purified through DEAE-52 anion exchange column and identified by dot-immunofiltration and immunoblotting assay with monoclonal antibody against cattle typeⅡcollagen.The purified CII of Z.dhumnades 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-1βand 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.dhumnades was 86.67%.In rats with arthri- tis,the ratio of CD4/CD8 subset and serum level of antibodies against CII and TNF-αwere increased,but that of IL-10 was de- creased.Meanwhile,the serum levels of IL-1βand IL-4 demonstrated no significant difference.These results suggest that typeⅡcollagen of Z.dhumnades(Cantor)can induce arthritis in rats with appearance of autoimmune reactions in vivo.更多还原