C57BL/6小鼠实验性变态反应性脑脊髓炎模型的建立

Establishment of Experimental Allergic Encephalomyelitis Model in C57BL/6 Mice

目的为更好地研究多发性硬化症(MS)的发病机理、开发具有良好效果的MS治疗药物,建立起能高度模拟人类多发性硬化症发生发展的小鼠模型则显得尤为必要且具有极高的应用价值。本文使用已广泛应用的标准化C57BL/6小鼠作为实验对象,成功地构建了实验性变态反应性脑脊髓炎模型(EAE)。此模型为研究人类多发性硬化症的发病机理及开发相关临床治疗药物提供了良好的小鼠模型。方法 C57BL/6雄性小鼠20只,分为2组。EAE组10只,皮下注射混匀的弗氏不完全佐剂与MOG53-58及结核分枝杆菌的悬浊液,同时分别在day0和day2腹腔注射百日咳毒素0.2μg/只;正常对照组10只,仅注射生理盐水。采用HE染色方法观察小鼠组织学变化;免疫组织化学方法进一步研究小鼠病理变化。结果从抗原免疫后第10天开始EAE组小鼠陆续发生EAE临床症状且发病率达到100%,而正常对照组小鼠未发生任何EAE临床症状。组织病理学结果分析显示,EAE组小鼠脊髓及脑组织中出现大量炎性细胞浸润且脱髓鞘严重。结论本实验成功构建的EAE小鼠模型病程稳定且发病率高,可用于MS机制研究及MS防治药物的筛选。

The establishment of an Experimental Allergic Encephalomyelitis(EAE) model in C57 BL/6 mice provides a better understanding to the mechanism of multiple sclerosis(MS) and a development of new drugs with good effects for MS treatment. Our study applied ten C57 BL/6 mice to immunize with both MOG53-58 and Mycobacterium tuberculosis(MT) in incomplete Freund’s adjuvant(IFA). And then they were treated with 0.2μg/mice Pertussis toxin(PTX) at day0 and day2 by i.p. injection to induce EAE model. Ten C57 BL/6 mice were just treated with normal saline as control group. Histological change in central nervous were investigated by H&E staining, immunohistochemistry assay and Luxol fast blue(LFB)/Periodic Acid-Schiff(PAS) double staining. The result comprise the mice in EAE group started to develop clinical symptoms from 10 th day post immunization, and all mice developed EAE disease. But no mice in control group developed EAE disease. Histological analysis showed abundant inflammatory cells filtrated into brain and spinal cord. Thus, the established EAE model is stable with high incidence, and this model could be used to study the mechanism of MS and screen the drugs for the prevention and treatment of MS.