小鼠流行性乙型脑炎疾病进展中反应性星形胶质细胞活化模式的变化及干预  

Changes and intervention of reactive astrocyte activation patterns in the progression of Japanese encephalitis

作　　者：周子清 李婧 王小可 思越 朱旭东 雷迎峰 马宏炜 张芳琳 ZHOU Ziqing;LI Jing;WANG Xiaoke;SI Yue;ZHU Xudong;LEI Yingfeng;MA Hongwei;ZHANG Fanglin(Department of Microbiology,Basic Medical Science Academy,The First Affiliated Hospital,Air Force Medical University,Xi’an 710032,China;Department of Critical Care Medicine and Department of Anaesthesiology and Perioperative Medicine,The First Affiliated Hospital,Air Force Medical University,Xi’an 710032,China)

机构地区：[1]空军军医大学基础医学院微生物与病原生物学教研室,陕西西安710032 [2]空军军医大学第一附属医院麻醉与围术期医学科/重症医学科,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2024年第4期303-310,共8页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(82172272)。

摘　　要：目的 明确星形胶质细胞活化模式与流行性乙型脑炎疾病进展的关系。方法 首先通过足垫注射乙型脑炎病毒(JEV)构建流行性乙型脑炎小鼠模型,采用免疫荧光技术(IFA)检测JEV非结构蛋白3(NS3)在小鼠全脑的表达情况;进一步利用IFA、 RNA测序技术(RNA-seq)、实时定量PCR明确流行性乙型脑炎发病不同阶段星形胶质细胞活化模式的变化;最后,通过侧脑室注射鸢尾素(irisin)调控补体C3阳性A1型星形胶质细胞、 S100钙结合蛋白A10(S100A10)阳性的A2型星形胶质细胞活化比例,探索其是否可改善乙型脑炎模型小鼠的体质量、行为学评分及脑组织损伤情况。结果 流行性乙型脑炎模型组小鼠的M1、 M2等运动皮层脑区和海马组织中NS3蛋白显著增加。上述区域胶质原纤维酸性蛋白(GFAP)阳性星形胶质细胞的数目和体积较对照组也显著增加,其中JEV感染后与A1/A2型星形胶质细胞活化相关的基因均显著升高。侧脑室或者腹腔注射irisin虽不能改善流行性乙型脑炎预后,但可在一定程度上抑制A1型星形胶质细胞活化,减轻神经炎症。结论 JEV主要感染小鼠的运动皮质和海马神经元,且星形胶质细胞异常激活导致神经炎症损伤。Irisin可能抑制A1型星形胶质细胞激活,减轻JEV感染后的神经炎症。Objective To clarify the relationship between astrocyte activation patterns and disease progression in epidemic encephalitis B(Japanese encephalitis).Methods First,a mouse model of epidemic encephalitis B was constructed by foot-pad injection of Japanese encephalitis virus(JEV),and the expression of viral protein NS3 in different brain regions was detected by immunofluorescence assay(IFA).Next,IFA,RNA sequencing(RNA-seq)and real-time quantitative PCR(qRT-PCR)were used to clarify the changes in the astrocyte activation patterns at different stages of epidemic encephalitis B.Finally,intracerebroventricular administration of irisin was conducted to regulate the proportion of activation in complement C3-positive A1 astrocytes and S100A10-positive A2 astrocytes,investigating whether it could improve the body mass,behavioral scores,and brain tissue damage in a mouse model.Results NS3 protein was detected by IFA predominantly in the M1/M2 region of the motor cortex and the hippocampus.The number and volume of GFAP-positive astrocytes significantly increased in JEV-infected brain regions,in which the expression of multiple genes associated with A1/A2 astrocyte activation was significantly enhanced.Although intracerebroventricular or intraperitoneal injection of irisin did not improve the prognosis of epidemic encephalitis B,it inhibited the activation of A1 astrocytes and ameliorate neuroinflammation.Conclusion Neurons in the M1/M2 motor cortex and hippocampus are susceptible to JEV infection,in which the abnormal astrocyte activa tion contributes to the neuroinflammatory injury.Irisin administration may restrain A1 astrocyte activation and alleviate neuroinflammation following JEV infection.

关 键 词：乙型脑炎病毒 星形胶质细胞 A1型星形胶质细胞 A2型星形胶质细胞 C3 S100A10 

分 类 号：R512.32[医药卫生—内科学] R364.5[医药卫生—临床医学] R373.31