小胶质细胞介导的T细胞应答在大鼠实验性自身免疫性脑脊髓炎中的致病作用  

The pathogenic role of microglia-mediated T cell responses in experimental autoimmune encephalomyelitis in rats

作　　者：周翔鱼[1] 李娜[1] 王佳[1] 储晨晨 祁莉凤 杨世胜 华冰[1] 曹羿堃 ZHOU Xiangyu;LI Na;WANG Jia;CHU Chenchen;QI Lifeng;YANG Shisheng;HUA Bing;CAO Yikun(Center for Neurological Diseases,the First People’s Hospital of Shizuishan,Shizuishan 753200,China)

机构地区：[1]宁夏石嘴山市第一人民医院神经疾病研究中心,宁夏石嘴山753200

出　　处：《宁夏医学杂志》2023年第9期772-775,F0002,共5页Ningxia Medical Journal

基　　金：国家自然科学基金项目(81660212)。

摘　　要：目的探讨大鼠维持中枢神经系统(CNS)炎症的小胶质细胞、T细胞亚群之间的相互效应关系,以期部分解释实验性自身免疫性脑脊髓炎(EAE)中中枢神经系统(CNS)持续存在病理损伤的原因。方法用鼠源性髓鞘碱性蛋白MBP68-86抗原肽制备Lewis大鼠EAE模型,透射电子显微镜观察脑组织超微结构,免疫荧光染色、Western blot、流式细胞术和RT-qPCR检测外周血、脾脏、脑组织中相关分子的表达。结果MBP68-86抗原肽成功制备出EAE模型。流式细胞及免疫荧光分析发现EAE模型组外周血CD4^(+)T(28.01%)、CD8^(+)T(12.81%)细胞的数量及脾脏中MHCⅡ的表达明显上调(P<0.05)。同时,在中枢神经系统中小胶质细胞的CD86^(+)CD206^(-)M1样巨噬细胞的数量明显多于CD86^(-)CD206^(+)M2样巨噬细胞。Western blot和RT-qPCR结果显示EAE模型组小胶质细胞及脑组织中MHCI、IL-10、TGF-β、CD32b,Granzyme B、Perforin、FASL分子表达明显上调(P<0.05)。结论大鼠EAE免疫应答过程中,CNS存在一个由CTL和Th2细胞共同参与的免疫应答环路,这可能是维持MS/EAE时间-空间多发性的主要机制。Objective To investigate the interaction between microglia and T cell subsets that maintain central nervous system(CNS)inflammation,in order to partially explain the persistent pathological damage of the central nervous system(CNS)in experimental autoimmune encephalomyelitis(EAE).Methods EAE model of lewis rat was prepared by MBP 68-86 antigenic peptide of mouse myelin basic protein.The ultrastructure of brain tissue was observed by transmission electron microscope,and the expression of related molecules in peripheral blood,spleen and brain tissue was detected by immunofluorescence staining,western blot,flow cytometry and RT-PCR.Results EAE model was successfully prepared by MBP 68-86 antigenic peptide.Flow cytometry and immunofluorescence analysis showed that CD4^(+)T(28.01%)and CD8^(+)T(12.81%)cells and MHCⅡ expression in spleen were significantly up-regulated in EAE model group(P<0.05).At the same time,the number of CD86^(+)CD206^(-)M1-like macrophages was significantly more than cd86^(-)CD206^(-)M2-like macrophages in the microglia of the central nervous system.Western blot and RT-PCR results showed that the molecular expressions of MHCI,IL-10,TGF-β,CD32b,Granzyme B,Perforin and FASL were significantly up-regulated in EAE model group(P<0.05).Conclusion During the EAE immune response,the CNS has an immune response loop,involving both CTLs and Th2 cells,which may be the main mechanism by which the temporal-spatial multiplicity of multiple sclerosis(MS)/EAE is maintained.

关 键 词：实验性自身免疫性脑脊髓炎 TH细胞 CTL细胞 MHCI 

分 类 号：R285.5[医药卫生—中药学]