摘要
目的:探讨β1,4-半乳糖基转移酶-1(β4GalT1)和CD200R1相互作用对小胶质细胞炎性活化的调节作用及其分子机制。方法:免疫共沉淀技术结合Western Blot法研究β4GalT1和CD200R1在体内外的相互作用;细胞免疫荧光技术观察两者定位。构建小胶质细胞-神经元共培养体系,阻断CD200-CD200R1结合,RT-PCR技术和ELISA试剂盒检测BV2细胞炎性因子i NOS、CD86、IL-1β、TNF-α的表达变化,LDH试剂盒检测神经元损伤情况。结果:β4GalT1和CD200R1有相互作用且存在共定位。CD200-CD200R1阻断剂增强LPS诱导的小胶质细胞炎性活化和神经元损伤,CD200R144号位突变体增强炎症因子释放和神经元损伤。CD200R144号位调节CD200-CD200R1结合,该突变体影响CD200-CD200R1的正常结合。结论:β4GalT1通过调节CD200R1第44号位的N-糖基化修饰,进而调节小胶质细胞的炎性活化及其介导的神经元损伤。
Objective:To explore the regulative effect ofβ4Galt1-CD200R1 interaction on inflammatory activation of microglia and its molecular mechanism.Methods:Immunoprecipitation combined with Western blot was used to explore the interaction betweenβ4Galt1 and CD200R1 in vivo and in vitro;the co-localization ofβ4Galt1 and CD200R1 was observed by cellular immunofluorescence technique.To construct the microglia-neuron co-culture system and block the binding of CD200-CD200R1,the iNOS,CD86,IL-1βand TNF-αexpression level was detected by RT-PCR and ELISA,and the neuron damage was detected by the LDH kit.Result:β4Galt1 can interact and co-locate with CD200R1.CD200-CD200R1 blocker enhanced LPS induced inflammatory activation and neuronal injury.The 44 position mutant of CD200R1 not only enhanced inflammatory factor release and neuronal damage,but also regulated the binding of CD200-CD200R1.Conclusion:β4Galt1 regulates the inflammatory activation of microglia and neuronal damage through mediating the N-glycosylation modification at position 44 of CD200R1.
作者
杨杰
刘超
段程伟
YANG Jie;LIU Chao;DUAN Chengwei(Neurosurgery Department,Nantong Third Hospital Affiliated to Nantong University/the Third People’s Hospital of Nantong,Jiangsu 226006;Central Lab,the First People’s Hospital of Wujiang Dist,Suzhou;Clinical Medical Research Center,Second Affiliated Hospital of Nantong University/the First People’s Hospital of Nantong)
出处
《交通医学》
2023年第2期111-116,共6页
Medical Journal of Communications
基金
南通市科技计划指导性项目(MSZ19110,JCZ20078)。
