DOCK2在缺血性卒中大鼠小胶质细胞除极和神经发生中作用研究.pdf

论著（收稿日期：2 0 2 3-0 1-10）503脑与神经疾病杂志2 0 2 3年第31卷第8 期参考文献1Graus F,Vogrig A,Muniz-Castrillo S,et al.Updated diagnosticcriteria for paraneoplastic neurologic syndromesJ.NeurolNeuroimmunol,2021,8(4):e1014.2 冉艳萍.儿童中枢神经系统副肿瘤综合征的研究进展.国际儿科学杂志，2 0 2 0,47(8):552-555.3 Grativvol RS,Cavalcante WCP,Castro LHM,et al.Updates in thediagnosis and treatment of paraneoplastic neurologic syndromesJ.Curr Oncol Rep.,2018,20(11):92.4Honnorat J,Didelot A,Karantoni E,et al.Autoimmune limbicencephalopathy and anti-Hu antibodies in children without cancerJ.Neurology,2013,80(24):2226.5 Chefdeville A,Treilleux I,Mayeur ME,et al.Immunopathologicalcharacterization of ovarian teratomas associated with anti-N-methyl-D-aspartate receptor encephalitisJJ.Acta Neuropathol Com,2019,7(1):38.6Jiang XY,Lei S,Zhang L,et al.Co-expression of NMDA-receptorsubunits NR1,NR2A,and NR2B in dysplastic neurons of teratomasin patients with paraneoplastic NMDA-receptor-encephalitis:a retrospective clinico-pathology study of 159 patientsJ.ActaNeuropathol Com,2020,8(1):130.7Mitchell WG,Blaes F.Cancer and autoimmunity:paraneoplasticneurological disorders associated with neuroblastic tumorsJJ.SeminPediatr Neurol,2017,24(3):180-188.8Dai Y,Zhang J,Ren H,et al.Surgical outcomes in patients with anti-N-methyl D-aspartate receptor encephalitis with ovarian teratomaJ.Am J 0Obstet Gynecol,2019,221(5):485.e1-485.e10.9Swayne A,Warren N,Prain K,et al.Analysing triggers for anti-NMDA-receptor encephalitis including herpes simplex virusencephalitis and ovarian teratoma:results from the Queenslandautoimmune encephalitis cohortJ.Intern Med J,2021,52(11):1943-1949.10Bost C,Chanson E,Picard G,et al.Malignant tumors in autoimmuneencephalitis with anti-NMDA receptor antibodiesJ.J Neurol,2018,265(10):2190-2200.11刘梦佳，邹丽萍.抗N-甲基-D-天冬氨酸受体脑炎与卵巢畸胎瘤相关性研究.中华神经医学杂志，2 0 15,14(9):9 7 0-9 7 1.12de Alarcon PA,Matthay KK,London WB,et al.Intravenousimmunoglobulin with prednisone and risk-adapted chemotherapyfor children with opsoclonus myoclonus ataxia syndrome associatedwith neuroblastoma(ANBLOOP3):a randomised,open-label,phase 3trialJ.Lancet Child Adolesc Health,2018,2(1):25-34.13 Wilbur C,Yea C,Licht C,et al.An upfront immunomodulatorytherapy protocol for pediatric opsoclonus-myoclonus syndrome.Pediatr Blood Cancer,2019,66(8):e27776.14 Iemura Y,Yamada Y,Hirata M,et al.Histopathologicalcharacterization of the neuroglial tissue in ovarian teratoma associatedwith anti-N-methyl-D-aspartate(NMDA)receptor encephalitisJ.Pathol Int,2018,68(12):677-684.16Guan HZ,Ren HT,Cui LY.Autoimmune encephalitis:anexpanding frontier of neuroimmunologyJJ.Chin Med J(Engl),2016,129(9):1122-1127.DOCK2在缺血性卒中大鼠小胶质细胞除极和神经发生中作用研究张华霖费长东毛琪刘慎周大伟【摘要】目的探究靶向胞质分裂作用因子蛋白（dedicatorof cytokinesis2DOCK2）蛋白在缺血性卒中（IS）大鼠脑组织中对小胶质细胞除极与神经发生的影响。方法将大鼠随机分为假手术（Sham）组，缺血性卒中（MCAO）组，IS+LC-shNC（M CA O+LV-s h NC）组，IS+LV-shDOCK2（M CA O+LV-s h D O CK 2）基金项目：辽宁省科学技术基金（2 0 19-ZD-1076）作者单位：116 0 0 0辽宁，中国人民解放军联勤保障部队第九六七医院急诊医学科（张华霖、费长东、毛琪、周大伟），神经外科（刘慎）通信作者：周大伟，Email：d r a g o n 7 6 10 2 8 12 6.c o m504脑与神经疾病杂志2 0 2 3年第31卷第8 期组，每组15只。7 d后开展神经功能评分与Y型迷宫检测，取脑组织进行TTC染色、免疫荧光染色，取缺血侧海马组织开展RT-PCR检测、Westernblot检测。结果与Sham组相比，MCAO组与MCAO+LV-shNC组大鼠脑组织中DOCK2表达增加（P0.05），出现梗死区域，神经功能评分增加（P0.05）。Y型迷宫中正确反应次数减少（P0.05），正确反应时间增加（P0.05）。此外，与Sham组相比，MCAO组与MCAO+LV-shNC组大鼠海马DG区BrdU阳性细胞数、DCX表达增加（P0.05），小胶质细胞活化标记物ibal表达升高（P0.05）。小胶质细胞极化标记物中，MCAO组与MCAO+LV-shNC组CD16、T NF-、IL-1、i NO SmRNA 表达明显增加（P0.05），且CD206、T CF-、A r g l、Yml mRNA 表达增加。然而，与MCAO组相比，MCAO+LV-shDOCK2组大鼠脑组织中DOCK2表达降低（P0.05），伴随着梗死面积减少（P0.05），神经功能评分减少（P0.05）。Y型迷宫中正确反应次数增加（P0.05），正确反应时间减少（P0.05）。与此同时，海马DG区BrdU阳性细胞数、DCX表达进一步增加（P0.05），但ibal表达降低（P0.05）。此外，M1型小胶质细胞标记物CD16、T NF-、IL-1、i NO SmRNA 表达减少（P0.05），M2型TGF-、A r g l、Ym l m RNA 表达进一步增加（P0.05）。结论敲低DOCK2蛋白表达有利于抑制小胶质细胞活化，逆转M1型小胶质细胞除极，提高M2型小胶质细胞除极化，最终起到促进神经发生，发挥神经功能与学习、记忆能力改善作用。【关键词】DOCK2；缺血性卒中；小胶质细胞除极；神经发生；神经功能；学习和记忆能力中图分类号：R743.32文献标识码：A文章编号：10 0 6-351X（2 0 2 3）0 8-0 50 3-0 8The role of DOCK2 in microglia polarization and neurogenesis in ischemic stroke ratsZhang Hualin,Fei Changdong,Mao Qi,Liu Shen,Zhou DaweiDepaitment of Emergency Medical,the 967 Hospital of the Joint Service Support Force of the Chinese Peoples LiberationArmy,Liaoning 116000,ChinaCorresponding author:Zhou Dawei,Email:Abstract Objective To investigate the effect of DOCK2 protein on microglial polarization andneurogenesis in rats with ischemic stroke(IS).Methods All rats were randomly divided into Group Shamoperation(Sham),Group IS(MCAO),Group ischemic stroke+LC-shNC(MCAO+LV-shNC),Group IS+LV-shDOCK2(MCAO+LV-shDOCK2),15 in each group.7d later,neurological function test and Y-maze test werecarried out,brain tissue was collected for TTC staining and immunofluorescence staining,and hippocampus from ISside was collected for RT-PCR and Western blot.Results Compared with Group Sham,the expression of DOCK2in the brain of the rats in Group MCAO and Group MCAO+LV-shNC increased,the infarct area appeared,and theneurological function score increased(P0.05).In the Y-maze,the number of correct response times decreased(P0.05),a