双特异性磷酸酶-1对小胶质细胞极化及颞叶癫痫发作的影响  

Effect of dual-specificity phosphatase-1 on microglial polarization and seizures in temporal lobe epilepsy

作　　者：张靖 张宇 任倩玉 安志恒 王丽敏 ZHANG Jing;ZHANG Yu;REN Qianyu;AN Zhiheng;WANG Limin(Department of Neurology,the First Hospital of Zhangjiakou,Zhangjiakou,Hebei 075000,China;Department of Outpatient,the 81st Group Army Hospital,Zhangjiakou,Hebei 075000,China)

机构地区：[1]河北省张家口市第一医院神经内科,河北张家口075000 [2]陆军第八十一集团军医院门诊部,河北张家口075000

出　　处：《检验医学与临床》2024年第15期2230-2236,共7页Laboratory Medicine and Clinic

基　　金：张家口市重点研发计划项目(1921091D)。

摘　　要：目的探讨双特异性磷酸酶-1(DUSP1)对小胶质细胞极化及颞叶癫痫(EP)发作的影响。方法采用慢病毒转染技术构建DUSP1正常表达组(DUSP1^(normal)组)、DUSP1高表达组(DUSP1^(over)组)、DUSP1低表达组(DUSP1^(low)组)小鼠,每组12只,将这些小鼠又分为DUSP1^(normal)+对照(Con)组、DUSP1^(normal)+EP组、DUSP1^(over)+Con组、DUSP1^(over)+EP组、DUSP1^(low)+Con组和DUSP1^(low)+EP组,每组6只。取DUSP1^(normal)+EP组、DUSP1^(over)+EP组及DUSP1^(low)+EP组小鼠分别腹腔注射氯化锂溶液(125 mg/kg)、硫酸阿托品溶液(1 mg/mL)及盐酸匹罗卡品溶液(50 mg/kg),构建颞叶EP模型。评估EP小鼠注射后2 h内EP发作等级,记录达到Ⅳ级的时间为潜伏期。将EP小鼠处死,取完整脑组织用于免疫荧光检测,取颞叶组织用于炎症细胞因子、DUSP1及小胶质细胞调控蛋白检测。结果与DUSP1^(normal)组比较,DUSP1^(over)组DUSP1/β-actin及DUSP1 mRNA表达水平均升高,DUSP1^(low)组DUSP1/β-actin及DUSP1 mRNA表达水平均降低,差异均有统计学意义(P<0.05)。与DUSP1^(normal)+EP组比较,DUSP1^(over)+EP组小鼠EP发作达Ⅳ级潜伏期明显延长,DUSP1^(low)+EP组小鼠EP发作达Ⅳ级潜伏期明显缩短,差异均有统计学意义(P<0.05)。与DUSP1^(normal)+Con组比较,DUSP1^(normal)+EP组小鼠颞叶组织中M1型小胶质细胞极化标志物CD16、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β及诱导型一氧化氮合酶(iNOS)mRNA水平均升高,炎症细胞因子TNF-α、IL-6水平均升高,IL-10水平降低,DUSP1诱导小胶质细胞极化相关蛋白p-胞外调节蛋白激酶(ERK)1/2、p-c-Jun氨基末端激酶(JNK)和p-p38蛋白表达水平均升高,差异均有统计学意义(P<0.05)。与DUSP1^(normal)+EP组比较,DUSP1^(over)+EP组小鼠颞叶组织中M1型小胶质细胞极化标志物CD16、TNF-α、IL-1β和iNOS mRNA水平均降低,M2型小胶质细胞极化标志物CD206、转化生长因子-β、精氨酸酶1与几丁质酶样蛋白mRNA水平均升高,炎症细胞因�Objective To investigate the effect of dual-specificity phosphatase-1(DUSP1)on microglia polarization and seizures in temporal lobe epilepsy(EP).Methods Lentivirus transfection technology was used to construct DUSP1^(normal) expression group(DUSP1^(normal) group),DUSP1 high expression group(DUSP1^(over) group),and DUSP1^(low) expression group(DUSP1^(low) group),with 12 mice in each group.These mice were divided into DUSP1^(normal)+control(Con)group,DUSP1^(normal)+EP group,DUSP1^(over)+Con group,DUSP1^(over)+EP group,DUSP1^(low)+Con group and DUSP1^(low)+EP group,with 6 mice in each group.The mice in the DUSP1^(normal)+EP,DUSP1^(over)+EP,and DUSP1^(low)+EP groups were intraperitoneally injected with lithium chloride solution(125 mg/kg),atropine sulfate solution(1 mg/mL),and pilocarpine hydrochloride solution(50 mg/kg)respectively,to establish a model of EP in the temporal lobe.The EP seizure grade of EP mice within 2 h after injection was evaluated,and the time to reach gradeⅣwas recorded as the latency period.The EP mice were sacrificed,and the intact brain tissues were collected for immunofluorescence detection,and the temporal lobe tissues were collected for the detection of inflammatory cytokines,DUSP1,and microglia regulatory proteins.Results Compared with the DUSP1^(normal) group,the expression levels of DUSP/β-actin and DUSP1 mRNA in the DUSP1^(over) group were significantly increased,while the expression levels of DUSP/β-actin and DUSP1 mRNA in the DUSP1^(low) group were significantly decreased(P<0.05).Compared with the DUSP1^(normal)+EP group,the DUSP1^(over)+EP group had a significantly longer latency to gradeⅣEP attack,and the DUSP1^(low)+EP group had a significantly shorter latency to gradeⅣEP attack(P<0.05).Compared with the DUSP1^(normal)+Con group,the mRNA levels of M1 microglial polarization markers CD16,tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand inducible nitric oxide synthase(iNOS)in the temporal lobe of the DUSP1^(normal)+EP group were increased,the levels of inflammatory cy

关 键 词：双特异性磷酸酶-1 小胶质细胞 丝裂原激活蛋白激酶 慢病毒载体 炎症反应 癫痫 

分 类 号：R742.1[医药卫生—神经病学与精神病学] R446.1[医药卫生—临床医学]