实验性自身免疫性脑脊髓炎小鼠模型的建立及其病理学和影像学表现

目的：建立实验性自身免疫性脑脊髓炎（experimental autoimmune encephalomyelitis,EAE）小鼠模型并做病理学及影像学鉴定,为研究多发性硬化的发病机制奠定基础。方法：C57BL/6雌性小鼠80只,分为EAE1组、EAE2组、EAE3组、EAE4组,各15只,用MOG35-55抗原加完全弗氏佐剂不同注射方法制作EAE小鼠模型;佐剂组10只用生理盐水代替MOG35-55制备免疫乳剂,对照组10只仅注射生理盐水;用MRI对小鼠脑和脊髓进行扫描,HE、LFB染色和Bielschowskys银染观察小鼠脑和脊髓病理学改变。结果：免疫后第14天,EAE1组、EAE2组、EAE3组、EAE4组小鼠开始出现EAE临床症状,发病率分别为80%、80%、73%、67%;EAE2组、EAE3组小鼠临床症状评分与EAE1组比较,P〉0.05,EAE4组小鼠临床症状评分与EAE1组比较,P〈0.05;佐剂组与对照组小鼠均未出现临床神经系统症状;EAE小鼠脑和脊髓小血管周围炎细胞浸润呈袖套状或脱髓鞘改变,神经元变性及轴突损伤,MRI可显示脑和脊髓病灶。结论：应用背部、颈部、腋窝、蹊部多点注射可提高EAE发病率,1次免疫与两次免疫小鼠对EAE发病率的影响没有明显差别。

Mouse Model Establishment and Pathological and Imaging Appearance of Autoimmune Encephalomyelitis

Objective：To establish autoimmune encephalomyelitis（AE）mouse model and observe its pathological and imaging appearance,and so as to provide a basis for study of pathological mechanism and therapy of multiple sclerosis.Methods：Eighty C57BL/6 female mice were divided randomly into 6 groups,with 15 in each of groups of EAE1,EAE2,EAE3,and EAE4.Experimental autoimmune encephalomyelitis（EAE）were induced in mice of the 4 groups with 300μg of MOG35-55 plus complete Freund＇s adjuvant（CFA）by different methods.Mice in adjuvant group（group ADJ,n=10）were treated with CFA plus normal saline,while mice in control group（group CON,n=10）treated with normal saline only.MRI examination and pathological observation on the brain and spinal cord were carried out on mice in groups of EAE1,ADJ,and CON.Methods used in pathological observation included HE staining,Luxol Fast Blue staining,and Bieschowsky silver staining.Results：Clinical symptoms of EAE appeared in mice of groups EAE1,EAE2,EAE3,and EAE4 in 14 days after immunization,and the incidences were 80%,80%,73%,and 67% respectively.Clinical symptom scores of groups EAE2 and EAE3 were not statistically different from that of group EAE1（P0.05）,while that of group EAE4 had statistically significant difference from that of group EAE1（P0.05）.No obvious EAE symptom was found in groups ADJ or CON.Pathological observation showed inflammatory cuff around small blood vessels,de-myelization,degeneration of neurons,and axonal injury in brain and spinal cord of mice in EAE groups.Foci of disease in brain and spinal cord were showed by MRI.Conclusions：Multi-point injection at back,neck,armpit,and groin could increase the incidence of EAE.Twice injection of reagent does not make significant difference from one single injection in the incidence of EAE.