重组人粒细胞集落刺激因子对实验性自身免疫性脑脊髓炎小鼠的治疗作用

摘要

Objective The purpose of this study is to explore the therapeutic effect of recombinant human granulocyte colony stimulating factor (rhG-CSF) on experimental autoimmune encephalomyelitis (EAE) mice. Methods Forty mice were randomly divided into 2 groups, and 10 in the control group were not treated, 30 in the model of EAE mice group induced by myelin oligodendrocyte glycoprotein 35-55, 6 mice were died, and the remaining 24 mice were randomly divided into model group and treatment group, with 12 mice in each group. The treatment group received rhG-CSF subcutaneously, and the model group received the same amount of saline. Neurological function score was evaluated during treatment. After 2 weeks of treatment, the brain and spinal cord specimens were taken for pathological observation and luxol fast blue (LFB) staining used for demyelination. The changes of the proportion of regulatory T (Treg) cells and natural killer (NK) cells were detected by flow cytometry. The expression of glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP) in EAE mice were detected by Western blot. Results The results showed that the neurological function score, the inflammatory cell infiltration score and the demyelinating score of the rhG-CSF treatment group mice were reduced. The proportion of Treg cells in the treatment group were significantly lower than that in the model group, and the proportion of NK cells in treatment group were significantly lower than that in model group. Western blot showed that the expression of GFAP in the treatment group was significantly lower than that in the model group, whereas the expression of MBP was significantly higher than that in the model group. Conclusion rhG-CSF has a significant therapeutic effect on EAE.%目的 观察重组人粒细胞集落刺激因子(recombinant human granulocyte colony stimulating factor, rhG-CSF)对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)小鼠的治疗效果并探讨相关机制.方法 将40只小鼠随机分为2组,对照组10只不作处理,30只用髓鞘少突胶质细胞糖蛋白35-55(纯度＞95％)多肽诱发建立EAE小鼠模型,死亡6只,剩余24只随机分为模型组和治疗组,各组12只,治疗组皮下注射rhG-CSF,模型组给予同等量的生理盐水.治疗期间对3组小鼠进行神经功能评分和体重检测;治疗2周后,取脑和脊髓制作标本行病理观察,劳克坚劳蓝染色进行脱髓鞘评分;流式细胞仪检测调节性T细胞和自然杀伤细胞比例的变化;蛋白质印迹法检测EAE小鼠胶质纤维酸性蛋白和髓鞘碱性蛋白的表达.结果 与模型组相比,治疗组小鼠神经功能评分显著降低,髓炎性细胞浸润评分和脱髓鞘评分降低;调节性T细胞显著增加,自然杀伤细胞显著减少;蛋白质印迹法的结果提示治疗组小鼠胶质纤维酸性蛋白表达量下降,而髓鞘碱性蛋白表达量明显高于模型组.结论rhG-CSF对EAE具有显著治疗效果.