免疫检查点受体CD200R1缺失激活小胶质细胞在帕金森病模型小鼠中的作用  

The role of microglia activated by the deletion of immune checkpoint receptor CD200R1 gene in a mouse model of Parkinson's disease

作　　者：郭佳丽 黄焘颖 张震 牛坤 巩比开·夏尔巴特 宫晓丽[2,3] 王晓民[2,3] 张婷[1,2] GUO Jia-Li;HUANG Tao-Ying;ZHANG Zhen;NIU Kun;GONGBIKAI Xarbat;GONG Xiao-Li;WANG Xiao-Min;ZHANG Ting(Department of Neurobiology,School of Basic Medical Sciences,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China;Key Laboratory of Neurodegenerative Disorders of the Ministry of Education,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China;Department of Physiology and Pathophysiology,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学基础医学院神经生物学系,北京100069 [2]首都医科大学教育部神经变性病重点实验室,北京100069 [3]首都医科大学基础医学院生理与病理生理学系,北京100069

出　　处：《生理学报》2025年第1期13-24,共12页Acta Physiologica Sinica

基　　金：National Natural Science Foundation of China(No.32171158,31971094);Beijing Natural Science Foundation(No.7222005);Scientific Research Common Program of Beijing Municipal Commission of Education(No.KM202010025032)。

摘　　要：本研究旨在探究维持小胶质细胞稳态的免疫检查点受体CD200R1缺失对小胶质细胞激活以及帕金森病(Parkinson's disease,PD)发病进程中黑质多巴胺(dopamine,DA)神经元丢失的影响。应用CRISPR-Cas9技术构建CD200R1-/-小鼠,提取野生型和CD200R1^(-/-)小鼠原代小胶质细胞,经脂多糖(lipopolysaccharide,LPS)处理后,用荧光微球吞噬实验检测小胶质细胞吞噬水平。小鼠黑质立体定位注射携带人源α-突触核蛋白(α-synuclein,α-syn)重组腺相关病毒载体制备PD模型,通过碰壁、旷场、转棒测试评估两种基因型小鼠运动行为学变化;用免疫组织化学染色标记DA神经元,评估DA神经元丢失情况;用免疫荧光染色检测小胶质细胞CD68(吞噬作用的关键分子)表达水平。结果显示,在体外,CD200R1缺失增强LPS诱导的小胶质细胞吞噬水平。CD200R1缺失加剧PD模型小鼠运动行为学损伤和黑质DA神经元丢失。在PD模型小鼠中,CD200R1-/-小鼠黑质小胶质细胞CD68荧光强度显著升高。以上结果提示,CD200R1缺失可能通过促进小胶质细胞吞噬功能进一步激活小胶质细胞,导致PD模型小鼠黑质DA神经元丢失加剧。CD200R1有望成为早期干预PD进程的新潜在靶点。The study aimed to investigate the effect of the CD200R1 gene deletion on microglia activation and nigrostriatal dopamine neuron loss in the Parkinson's disease(PD)process.The CRISPR-Cas9 technology was applied to construct the CD200R1_(−/−)mice.The primary microglia cells of wild-type and CD200R1−/−mice were cultured and treated with bacterial lipopolysaccharide(LPS).Microglia phagocytosis level was assessed by a fluorescent microsphere phagocytosis assay.PD mouse model was prepared by nigral stereotaxic injection of recombinant adeno-associated virus vector carrying humanα-synuclein(α-syn).The changes in the motor behavior of the mice with both genotypes were evaluated by cylinder test,open field test,and rotarod test.Immunohistochemical staining was used to assess the loss of dopamine neurons in substantia nigra.Immunofluorescence staining was used to detect the expression level of CD68(a key molecule involved in phagocytosis)in microglia.The results showed that CD200R1 deletion markedly enhanced LPS-induced phagocytosis in vitro by the microglial cells.In the mouse model of PD,CD200R1 deletion exacerbated motor behavior impairment and dopamine neuron loss in substantia nigra.Fluorescence intensity analysis results revealed a significant increase in CD68 expression in microglia located in the substantia nigra of CD200R1−/−mice.The above results suggest that CD200R1 deletion may further activates microglia by promoting microglial phagocytosis,leading to increased loss of the nigrostriatal dopamine neurons in the PD model mice.Therefore,targeting CD200R1 could potentially serve as a novel therapeutic target for the treatment of early-stage PD.

关 键 词：CD200R1 小胶质细胞 吞噬 帕金森病 

分 类 号：R74[医药卫生—神经病学与精神病学]