目的比较不同剂量髓鞘少突胶质细胞糖蛋白(myelin oligodendrocyte glycoprotein,MOG)35-55免疫诱导C57BL/6小鼠实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)的作用以及相关免疫组织学观察。方法将C57BL/6小鼠分为正常组和三个不同剂量MOG35-55诱导的EAE模型组,共4组。模型组分别以每只300、150、50μg的MOG35-55与等量完全弗氏佐剂(complete Freund's adjuvant,CFA)混合的乳化抗原皮下注射诱导EAE模型,正常组以生理盐水代替。观察不同剂量MOG35-55对C57BL/6小鼠体重、发病率、死亡率以及神经功能评分等的影响,同时HE染色观察小鼠脑和脊髓组织神经病理学改变以及免疫组化法观察脑和脊髓组织少突胶质细胞转录因子2(oligodendrocyte transcription factor 2,Olig2)、髓鞘碱性蛋白(myelin basic protein,MBP)、神经胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)的表达情况。结果三组不同剂量MOG35-55均能诱导EAE模型,呈慢性单相病程,病理学观察发现小鼠脑和脊髓组织有炎性细胞浸润、脱髓鞘等改变。但小剂量组在体重减轻、神经功能评分及病理学改变、大脑及脊髓特定部位Olig2、MBP、GFAP表达等方面均较其他模型组明显。结论用MOG35-5550μg免疫诱导的C57BL/6小鼠EAE模型稳定可靠,可以作为研究多发性硬化(multiple sclerosis,MS)的理想模型。
Objective To compare the conditions to induce experimental autoimmune encephalomyelitis(EAE)in C57BL /6 mice with myelin oligodendrocyte glycoprotein35-55 and the pathological characteristics revealed by immunohisto-chemistry.Methods Fifty female SPF C57BL /6 mice 6-8 weeks old were divided randomly into four groups:normal group and three EAE model groups(high-dose group,middle-dose group and low-dose group).The mice of the three model groups were injected subcutaneously over flanks with the antigen containing 300,150,50 μg MOG35-55 /mouse and completeFreund's adjuvant(CFA) in the same ratio,respectively.500 ng pertussis toxin(PTX) in 200 μL physiological saline was given i.p.to the mice of the three model groups on the day of immunization and 48 h later.Physiological saline was given to the normal group instead.The effects of different doses of MOG35-55 on the body weight,incidence,mortality rate and neu-rological scores of C57BL /6 mice were observed.Meanwhile,the pathological changes of CNS were examined by hematoxy-lin-eosin(HE) staining and the expression of Olig2,MBP and GFAP by immunohistochemical staining.Results The C57BL /6 mouse model of EAE was successfully induced with three different doses of MOG35-55.The EAE model represen-ted a chronic,monophasic feature of disease.There were inflammatory cell infiltration around small veins called sleeve-like changes and demyelination in the brain and spinal cord of mice.However,the influence of low-dose of MOG35-55 on loss of weight,neurological score and pathological changes as well as the expression of Olig2,MBP and GFAP seemed to be more significant than that in other model groups.Conclusion An immune-induced mouse model of EAE has been successfully established with MOG35-55,especially with a low dose of 50 μg.This EAE mouse model is stable,with a high incidence and low mortality rate,and can be used in research of multiple sclerosis(MS).