小胶质细胞在PLP139-151诱导SJL小鼠EAE模型中的作用研究  

Microglia in PLP139-151-induced EAE model in SJL mice

作　　者：丛衡日 张明[2] 苌浩晓 杜利 尹琳琳[2] 张星虎[1] CONG Hengri;ZHANG Ming;CHANG Haoxiao;DU Li;YIN Linlin;ZHANG Xinghu(Department of Neurology,Beijing Tiantan Hospital,Capital Medical University,China National Clinical Research Center for Neurological Diseases,Beijing 100070,China)

机构地区：[1]首都医科大学附属北京天坛医院神经病学中心,国家神经系统疾病临床医学研究中心,100070 [2]首都医科大学宣武医院药物研究室,100053

出　　处：《中国神经免疫学和神经病学杂志》2019年第6期405-409,共5页Chinese Journal of Neuroimmunology and Neurology

基　　金：北京市自然科学基金面上项目(7162208);北京市卫生系统高层次人才(学科骨干,2014-3-052)。

摘　　要：目的观察髓鞘蛋白脂质蛋白139-151(proteolipid protein,PLP139-151)诱导SJL小鼠构建的复发缓解型的实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)模型复发期脑内小胶质细胞的病理改变。方法应用100μg、150μg PLP139-151多肽片段免疫SJL小鼠构建复发缓解型EAE模型(分别简称为100μg PLP组、150μg PLP组),另设健康对照组(对照组;采用与实验组等体积溶媒处理),观察35 d。观察两组EAE小鼠的发病情况及神经功能残疾进展评分差异,选择发病快、神经功能残疾进展评分高者(计为EAE组)观察复发期小胶质细胞活化情况。EAE组于复发期疾病高峰时,对照组于相同时间点,取小鼠大脑皮层组织,应用免疫组织化学(immunohistochemical,IHC)染色及免疫荧光(immunofluorescence,IF)染色比较两组小胶质细胞的活化情况。结果150μg PLP组小鼠在两个发病高峰(分别为免疫第13、28天)均早于100μg PLP组(分别为免疫第15天、31天),神经功能评分第2次发病高峰高于100μg PLP组(3.00±0.79比1.75±0.90,t=2.33,P<0.05),第1次发病高峰与100μg PLP组差异无统计学意义(P>0.05),故选择150μg PLP组(计作EAE组)观察复发期(免疫后第28天)小胶质细胞活性。与对照组相比,EAE组小鼠复发期大脑皮层中小胶质细胞的活化明显增多,组织染色上表现为Iba-1表达增高,细胞体积增大,细胞突起增多。结论150μg PLP139-151多肽片段免疫SJL小鼠构建的复发缓解型EAE模型复发期小鼠大脑皮层小胶质细胞明显活化。Objective To observe the pathological changes of microglia in the rlapse period of experimental autoimmune encephalomyelitis(EAE)model in SJL mice induced by proteolipid protein(PLP139-151).Methods SJL mice were immunized with 100μg and 150μg PLP139-151 to construct a relapsing-remitting EAE model,and a healthy control group(the control group was treated with the same volume of solvent corresponding to the experimental group)was set up.All groups were observed for 35 days.The development of EAE and the progression of neurological disability were observed in both EAE groups.We chose the EAE group with a rapid onset and high score of neurological disability to observe the activation of microglia in the relapse period.The activation of microglia in the chosen EAE group was compared with that in the control group at the same time using immunohistochemically staining(IHC)and immunofluorescence staining(IF).Results the 150μg PLP induced EAE model reached disease peak(13th and 28th days after immunization)earlier than the 100μg PLP group(15th and 31th days after immunization),and its neurological function score was higher than the 100μg PLP group(3.00±0.79 vs.1.75±0.90,t=2.33,P<0.05)in the second disease peak.There was no significant difference of neurological function score between the two groups in the first disease peak period(P>0.05).So the 150μg PLP group was selected to observe the activity of microglia in the reapse period(28 days after immunization).Compared with the control group,the activation of microglia in the cerebral cortex of EAE group increased significantly,which presented as increased expression of Iba-1 cell volume,and cell dendrites.Conclusions The activation of microglia cells in the recurrent EAE model constructed by SJL mice immunized with 150μg PLP139-151 is obvious when compared with the control group.

关 键 词：实验性自身免疫性脑脊髓炎 多发性硬化 髓鞘蛋白脂质蛋白 小胶质细胞 

分 类 号：R744.52[医药卫生—神经病学与精神病学]