低氧减弱CD4^(+)T细胞激活降低脑脊髓炎小鼠神经病理损伤  

Hypobaric hypoxia alleviates the neuropathological injury in the experimental autoimmune encephalomyelitis mice by attenuating peripheral CD4^(+)T cell activation

作　　者：员鹏飞 张倩 董小铷 刘新秦 赵芳 孟姗姗 张文斌 陈筱鸣 Yuan Pengfei;Zhang Qian;Dong Xiaoru;Liu Xinqin;Zhao Fang;Meng Shanshan;Zhang Wenbin;Chen Xiaoming(The 18th Squadron,The 5th Battalion,School of Basic Medicine,School of Public Health,Air Force Military Medical University,Xi’an 710032,China;Department of Occupational and Environmental Health and the Ministry of Education,School of Public Health,Air Force Military Medical University,Xi’an 710032,China)

机构地区：[1]空军军医大学基础医学院五大队十八队,西安710032 [2]空军军医大学军事预防医学系军队劳动与环境卫生学教研室,西安710032

出　　处：《神经解剖学杂志》2021年第5期493-501,共9页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81803194)。

摘　　要：目的:分别于常氧环境和低压低氧环境下建立实验性自身免疫性脑脊髓炎(EAE)小鼠模型,通过测定小鼠的神经病理损伤和外周免疫激活,评估低压低氧环境对EAE小鼠的病理影响和潜在机制。方法:C57BL/6J雌性小鼠皮下注射髓鞘少突胶质细胞糖蛋白33-35(MOG33-35),建立EAE模型,于常氧环境和低压低氧环境(模拟海拔4500 m)分别暴露32 d,通过测定小鼠体重、临床病理评分(CS)评估小鼠病理损伤,利用苏木精-伊红(HE)染色测定中枢神经系统(CNS)炎性浸润、腊克沙固蓝(LFB)染色评估白质脱髓鞘状况,并利用CD3、CD4、IL-17α和IFN-γ免疫组织化学染色明确CNS浸润的免疫细胞种类;此外,利用酶联免疫吸附试验(ELISA)测定血清细胞因子IL-17α、IFN-γ和IL-21水平,流式细胞分析测定脾脏CD4^(+)效应T细胞中IL-17α^(+)、IFN-γ^(+)和IL-2^(+)细胞比例,进而评估小鼠外周免疫激活状态。结果:与常氧EAE组小鼠相比,低压低氧EAE组小鼠临床病理评分在疾病高峰期显著降低,同时HE染色和LFB染色显示CNS炎性浸润和白质脱髓鞘均显著减轻,提示小鼠神经病理损伤减轻。此外,低压低氧EAE小鼠CNS中CD3^(+)、CD4^(+)、IFN-γ^(+)细胞浸润显著减少,同时血清中IFN-γ和IL-17α细胞因子水平显著降低,脾脏激活的IL-17α^(+)、IFN-γ^(+)的CD4^(+)效应T细胞比例显著降低,提示低压低氧导致EAE小鼠外周免疫激活减弱,可能是中枢浸润免疫细胞减少的主要原因。结论:低压低氧环境可以通过减弱外周CD4^(+)T细胞的免疫激活,减少IFN-γ和IL-17α细胞因子的分泌,从而减少CNS免疫细胞浸润,降低EAE小鼠神经病理损伤。Objective:Experimental autoimmune encephalomyelitis(EAE)mice models were established under hypobaric hypoxia and normoxia,respectively.Neuropathological injury and peripheral immune activation were measured to evaluate the influence of hypobaric hypoxic exposure upon the EAE neuropathological development and explore the potential mechanism.Methods:Female C57BL/6J mice were injected subcutaneously with myelin oligodendrocyte glycoprotein 33-35(MOG33-35)to establish EAE models,and then exposed to normoxic and hypobaric hypoxic environment(simulated 4500 m altitude)for 32 days,respectively.Body weights and clinical scores(CS)were measured to evaluate the neuropathological development.hematoxylin-eosin(HE)and luxol fast blue(LFB)staining were used to assess the inflammatory infiltration and demyelination in the mice central nervous system(CNS).Immunohistochemical staining of CD3,CD4,IL-17α,and IFN-γwere used to identify the types of CNS-infiltrated immune cells.To assess peripheral CD4^(+)T cells activation,enzyme-linked immunosorbent assay(ELISA)was used to measure the seral cytokine levels(IL-17α,IFN-γ,and IL-21),and flow cytometry was used to measure the spleen effector CD4^(+)T cell subsets(IL-17α^(+),IFN-γ^(+),and IL-2^(+)).Results:Compared with the normoxic EAE mice,the clinical scores(at the peak of the disease)significantly reduced in the hypobaric hypoxic EAE mice.Similarly,the CNS inflammatory infiltration and demyelination in the hypobaric hypoxic EAE mice were also lower than those in the normoxic EAE mice,indicating that the neuropathological injury in the EAE mice were alleviated under hypobaric hypoxia.Among the CNS-infiltrated immune cells,the CD3^(+),CD4^(+),and IFN-γ^(+)cells significantly decreased under hypobaric hypoxia.As for the peripheral immune activation,the seral IFN-γand IL-17αlevels significantly reduced in the hypobaric hypoxic EAE mice,and the proportion of splenic IL-17α^(+),and IFN-γ^(+)effector CD4^(+)T cells also significantly decreased,indicating that the peripheral immu

关 键 词：低压低氧暴露 多发性硬化 实验性自身免疫性脑脊髓炎模型 神经病理损伤 外周免疫激活 小鼠 

分 类 号：R744.51[医药卫生—神经病学与精神病学]