Pristane诱导小鼠系统性红班狼疮动物模型的研究  

Study on experimental systemic lupus erythematosus mouse model induced by pristane

作　　者：李金枝[1] 丁隽[2] Merim Boukherouba 李昊文[1] 张勇[1] 沈浩[1] 陈学进[2] 王树军[1] 王颖[1] 

机构地区：[1]上海交通大学医学院上海市免疫学研究所,上海200025 [2]上海交通大学医学院实验动物科学部,上海200025

出　　处：《细胞与分子免疫学杂志》2011年第2期119-122,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家高技术研究发展计划(863)资助项目(2006AA02A252);上海市国际科技合作基金项目资助(10410701600);上海市科委基金资助项目(102R1426200);上海市教委第5期重点学科(J50207)

摘　　要：目的:建立pristane诱导的系统性红斑狼疮(SLE)小鼠模型,并对该小鼠模型的发病机制进行初步的探讨。方法:6-8周龄雌性BALB/c小鼠单次腹腔注射pristane0.5mL,对照组单次腹腔注射PBS0.5mL,注射前及注射后每2周行流式细胞术(FCM)检测外周血中IFN-α分泌细胞(CD11b+Ly6Chigh)的比例及细胞活化状态B220+Aβ1dhigh),ELISA检测血清中自身抗体(anti-dsDNA,anti-smRNP,anti-ribosomalP0)的含量。至6个月处死动物,FCM检测腹腔细胞中IFN-α分泌细胞(CD11b+Ly6Chigh)的比例和脾脏中细胞的活化(B220,Aβ1d),采用直接免疫荧光法标记小鼠肾脏免疫球蛋白复合物及H&E染色评估小鼠肾脏免疫复合物的沉积及损伤情况。结果:小鼠腹腔注射pristane第2个月开始血清总IgG升高,第3个月起出现自身抗体阳性,到个6月时达到最高,并维持高水平至被处死;Pristane处理6个月后,Pristane处理组小鼠出现关节炎症状,肾脏免疫复合物的大量沉积和明显肾脏损伤。Pristane注射2周起,小鼠外周血中IFN-α分泌细胞(CD11b+Ly6Chigh)的比例明显高于PBS注射组,小鼠腹腔细胞中IFN-α分泌细胞的比例也明显升高;同时外周血和脾细胞中B细胞表面MHCII分子Aβ1d的平均荧光强度(MFI)均高于对照组,表明pristane处理组小鼠中B细胞发生了显著活化。结论:BALB/c小鼠腹腔注射pristane可诱导构建小鼠SLE模型,其SLE的发病可能与IFN-α的持续分泌导致B细胞的异常活化有关。该模型的建立为进一步研究SLE的发病机制提供了良好的动物模型。AIM： To establish the systemic lupus erythematosus （SLE） mouse model through pristane intraperitoneal injection and discuss the pathogenesis of SLE in this mouse model. METHODS： Single intraperitoneal injection of 0.5 mL Pristane or PBS was applied on 6 - 8 week old female BALB/c mice. The percentage of IFN-α producing cells （CDllb^＋ Ly6C^high）and B cells and the expression of B cell activation surface marker （Aβ1^d） in peripheral blood were detected by flow cytometry every 2 weeks. Serum total IgG and auto-antibodies （anti-dsDNA, anti-sm RNP, anti-ri- bosomal P0）were detected by ELISA at different time point. The percentage of peritoneal CDIIb^＋ Ly6Ch^high cells and Aβ1^d expression in spleen were also detected by flow cytometry after 6 months, glomerular IgG deposition and kidney histopathologic changes were determined by direct immunofluorescence and H＆E staining respectively. RESULTS： Total IgG began to increase since 2 months after the pristane injection, while auto-antibodies were detected after 3 moths, both of which peaked after 6 moths and maintained the high level. Most of the pristane treated mice developed arthritis, glomerular immune complex deposition and kidney damage. The percentage of peripheral and peritoneal IFN-α producingcells was much higher in pristane group than that of the PBS control group since 2 weeks after intraperitoneal injection. The mean fluorescence intensity （MFI） of B cell activation marker（Aβ1^d） in pristane group was also higher than PBS group in both peripheral blood and spleen indicating B cell over activation. CONCLUSION. Intraperitoneal injection of pristane can successfully establish a SLE mouse model which may be used in research of the SLE pathogenesis. increased percentage of IFN-α producing cells may play an etiopathogenic role in abnormal B cell activation and SLE development in this model.

关 键 词：系统性红斑狼疮 小鼠模型 IFN-Α B细胞活化 自身抗体 

分 类 号：R392.33[医药卫生—免疫学]