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目的探讨实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)发病过程中淋巴细胞相关因子及转录因子的表达情况。方法将C56BL/6小鼠分为免疫髓鞘少突胶质细胞糖蛋白35-55(myelin oligodendrocyte glycoprotein 35-55,MOG35-55)特异性免疫乳剂的EAE组与免疫未加MOG35-55多肽的免疫乳剂的CFA组,每组16只,分别取100μl免疫乳剂,经腋窝皮下免疫小鼠,于免疫当天及第2天经小鼠尾静脉注射200 ng百日咳毒素(pertussis toxin,PT),建立实验动物模型,并于免疫后进行临床评分;分别于免疫后第7、14和21天取小鼠的淋巴结和脾脏,RT-PCR法检测淋巴细胞相关因子及转录因子IL-17、IFNγ、RAR相关孤核受体γ(RAR-related orphan receptor gamma,RORγ)和T盒子转录因子be(tT-box transcription factor bet,T-bet)基因mRNA转录水平的变化。结果成功建立EAE模型,从免疫后第7天起出现临床症状,EAE组临床评分明显高于CFA组(P均<0.05);免疫后第7、14和21天,EAE组淋巴结和脾脏淋巴细胞中IL-17、IFNγ、RORγ、T-bet基因mRNA转录水平均明显高于CFA组。结论 IL-17、IFNγ、RORγ和T-bet参与EAE疾病的发生发展,且表达量的增加与疾病加重有相关性。
Objective To investigate the expression of lymphocyte related factors and transcription factors in the pathogenesis of experimental autoimmune encephalomyelitis (EAE). Methods C56BL / 6 mice were divided into EAE group and immunized myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) Immunofluorescein CFA group, 16 mice in each group were immunized with 100μl immunosuppressive agents. The mice were immunized subcutaneously with axillas. 200 ng pertussis toxin (PT) was injected through the tail vein of mice on the day of immunization and on the second day to establish the experiment The animal model and clinical score after immunization. The lymph nodes and spleens of mice were taken on the 7th, 14th and 21th day after immunization. The correlation of lymphocyte related factors and the transcription factors IL-17, IFNγ, RAR was detected by RT-PCR Changes of mRNA transcription level of RAR-related gene and T-box transcription factor bet (T-bet transcription factor) mRNA. Results The EAE model was successfully established. Clinical symptoms were observed on the 7th day after immunization. The clinical scores of EAE group were significantly higher than those of CFA group (all P <0.05). On the 7th, 14th and 21st day after immunization, the lymph node and spleen lymphocytes The mRNA transcription levels of IL-17, IFNγ, RORγ and T-bet mRNA in CFA group were significantly higher than those in CFA group. Conclusion IL-17, IFNγ, RORγ and T-bet are involved in the occurrence and development of EAE disease, and the increase of expression level is correlated with the exacerbation of disease.