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  

The role of DOCK2 in microglia polarization and neurogenesis in ischemic stroke rats

作　　者：张华霖 费长东 毛琪 刘慎 周大伟 Zhang Hualin;Fei Changdong;Mao Qi;Liu Shen;Zhou Dawei(Depaitment of Emergency Medical,the 967 Hospital of the Joint Service Support Force of the Chinese People's Liberation Army,Liaoning 116000,China)

机构地区：[1]中国人民解放军联勤保障部队第九六七医院急诊医学科,辽宁116000 [2]中国人民解放军联勤保障部队第九六七医院神经外科,辽宁116000

出　　处：《脑与神经疾病杂志》2023年第8期503-510,共8页Journal of Brain and Nervous Diseases

基　　金：辽宁省科学技术基金(2019-ZD-1076)。

摘　　要：目的 探究靶向胞质分裂作用因子蛋白(dedicator of cytokinesis 2,DOCK2)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 (MCAO+LV-shNC)组,IS+LV-shDOCK2 (MCAO+LV-shDOCK2)组,每组15只。7d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、Western blot检测。结果 与Sham组相比,MCAO组与MCAO+LVshNC组大鼠脑组织中DOCK2表达增加(P<0.05),出现梗死区域,神经功能评分增加(P<0.05)。Y型迷宫中正确反应次数减少(P<0.05),正确反应时间增加(P<0.05)。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(P<0.05),小胶质细胞活化标记物iba1表达升高(P<0.05)。小胶质细胞极化标记物中,MCAO组与MCAO+LV-shNC组CD16、TNF-α、IL-1 β、iNOS mRNA表达明显增加(P<0.05),且CD206、TGF-β、Arg1、Ym1 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(P<0.05),伴随着梗死面积减少(P<0.05),神经功能评分减少(P<0.05)。Y型迷宫中正确反应次数增加(P<0.05),正确反应时间减少(P<0.05)。与此同时,海马DG区BrdU阳性细胞数、DCX表达进一步增加(P<0.05),但iba1表达降低(P<0.05)。此外,M1型小胶质细胞标记物CD16、TNF-α、IL-1 β、iNOS mRNA表达减少(P<0.05),M2型TGF-β、Arg1、Ym1 mRNA表达进一步增加(P<0.05)。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。Objective To investigate the effect of DOCK2 protein on microglial polarization and neurogenesis in rats with ischemic stroke(IS).Methods All rats were randomly divided into Group Sham operation(Sham),Group IS(MCAO),Group ischemic stroke+LC-shNC(MCAO+LV-shNG),Group IS+LVshDOCK2(MCAO+LV-shDOCK2),15 in each group.7d later,neurological function test and Y-maze test were carried out,brain tissue was collected for TTC staining and immunofluorescence staining,and hippocampus from IS side 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 the neurological function score increased(P<0.05).In the Y-maze,the number of correct response times decreased(P<0.05),and the correct response time increased(P<0.05).In addition,compared with Group Sham,the number of BrdU positive cells and the expression of DCX in the hippocampal DG area of the rats in Group MCAO and Group MCAO+LV-shNC increased(P<0.05),and the expression of the microglia activation marker ibal increased(P<0.05).Among the microglia polarization markers,the expressions of CD16,TNF-α,IL-1β,and iNOS mRNA in Group MCAO and Group MCAO+LV-shNC were significantly increased(P<0.05),as well as,the expressions of CD206,TGF-β,Argl,and Yml mRNA increased.However,compared with Group MCAO,the expression of DOCK2 in rats of Group MCAO+LV-shDOCK2 decreased(P<0.05),which accompanied with a decrease in the infarct size(P<0.05)and neurological score(P<0.05).In addition,compared with Group MCAO,in the Y-maze,the number of correct response times increased(P<0.05)and the correct response time decreased(P<0.05)of Group MCAO+LVshDOCK2.At the same time,the number of BrdU-positive cells and the expression of DCX in the hippocampal DG area were further increased,but the expression of ibal decreased of Group MCAO+LV-shDOCK2.Furthermore,the expressions of M1-type microglia markers CD16,TNF-α,IL-1β,and iNOS mRNA decreased(P<0.05),and the express

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 

分 类 号：R743.32[医药卫生—神经病学与精神病学]