Maresin 2对小鼠蛛网膜下腔出血早期的神经保护作用研究  

Research on the Neuroprotective Effects of Maresin 2 in the Early Stage of SubarachnoidHemorrhage in Mice

作　　者：李晨溪 刘一涓 王华飞 王莉[2] 余列[3] 臧卫东 LI Chenxi;LIU Yijuan;WANG Huafei;WANG Li;YU Lie;ZANG Weidong(Department of Human Anatomy,School of Basic Medicine,Zhengzhou University,Zhengzhou 450001,China;Biobank,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450001,China;Department of Neurology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450001,China)

机构地区：[1]郑州大学基础医学院人体解剖学系,郑州450001 [2]郑州大学第一附属医院生物样本库,郑州45000 [3]郑州大学第一附属医院神经内科,郑州450001

出　　处：《神经损伤与功能重建》2024年第6期311-316,共6页Neural Injury and Functional Reconstruction

基　　金：河南省科技攻关项目(Prdx1的新功能—调控脑梗死后SVZ区神经发生效率及其YTHDC1依赖机制,No.202102310076;CES2C介导p53烷基化抑制脑梗死小鼠少突胶质前体细胞衰老促进髓鞘再生的分子机制研究,No.242102311133)。

摘　　要：目的:探讨Maresin 2(Mar2)注射对蛛网膜下腔出血(SAH)小鼠早期的神经保护作用及机制。方法:将134只雄性C57BL/6小鼠按照随机数字表法分为Sham组(12只)、Vehicle组(22只)、Mar2组(57只)、牛磺熊去氧胆酸钠(Tau)组(29只)和Tau+Mar2组(14只),其中Mar2组又分为低(Low;2μg/kg/d;14只)、中(Middle;10μg/kg/d;29只)、高(High;50μg/kg/d;14只)剂量3个亚组。采用改良血管内穿刺法制备SAH模型,在SAH造模后6 h腹腔注射Mar2;Tau组,按照0.5 g/kg/d的剂量腹腔注射Tau,连续注射3 d;Tau+Mar2组,腹腔注射Tau后2 h,再腹腔注射中剂量的Mar2。完成注射后对各组小鼠进行神经功能评分、脑出血评分和脑组织含水量测定,FJB染色评估神经元变性数量,免疫荧光染色观察星形胶质细胞和小胶质细胞的数量变化,Western blotting检测神经元中内质网应激蛋白C/EBP同源蛋白(CHOP)和caspase-3的表达,荧光实时定量PCR(qPCR)检测脑组织白细胞介素-1β(IL-1β)、IL-6炎症因子的变化。体外培养Neuro2a神经元,根据实验采用血红素(Hem)或Tau(CHOP特异性抑制剂)进行干预,并分为对照组、Hem组、Hem+Tau组、Hem+Tau+Mar2组。Western blotting检测各组神经元表达CHOP和caspase-3水平;酶联免疫法检测活性氧(ROS)的水平。结果:Mar2注射显著改善神经功能缺损、减轻脑水肿,抑制神经元凋亡,中剂量(10μg/kg/d)即可达到最佳治疗效果。Western blotting结果显示Mar2可以抑制皮质CHOP和caspase-3的表达;免疫荧光染色结果显示,Mar2可以抑制星形胶质细和小胶质细胞的活化增多;qPCR检测结果显示,Mar2注射可显著减少炎症因子IL-1β、IL-6的表达。体外实验显示,血红素可诱导神经元中CHOP和caspase-3的显著上调和ROS的合成增多;Mar2处理可以显著抑制神经元CHOP和caspase-3的表达。高剂量Tau处理可以显著抑制神经元中CHOP和caspase-3的表达,Tau+Mar2处理不能进一步降低CHOP和caspase-3的表达。结论:Mar2注射能抑制SAObjective:To investigate the neuroprotective effects and mechanisms of Maresin 2(Mar2)in the early stage of subarachnoid hemorrhage(SAH)in mice.Methods:A total of 134 male C57BL/6 mice were randomly divided into Sham group(12 mice),Vehicle group(22 mice),Mar2 group(57 mice),Tau group(29 mice),and Tau+Mar2 group(14 mice),with the Mar2 group further divided into low(2μg/kg/d;14 mice),middle(10μg/kg/d;29 mice),and high(50μg/kg/d;14 mice)dose subgroups.The SAH model was prepared using a modified intravascular perforation method,and Mar2 was injected intraperitoneally 6 hours after SAH modeling;the Tau group received an intraperitoneal injection of Tau at a dose of 0.5 g/kg/d for 3 consecutive days;the Tau+Mar2 group received an intraperitoneal injection of Tau followed by a middle dose of Mar22 hours later.After the injections,neurological function scores,cerebral hemorrhage scores,and brain tissue water content were measured for each group of mice.FJB staining was used to assess the number of degenerating neurons,immunofluorescence staining was used to observe changes in the number of astrocytes and microglia,Western blotting was used to detect the expression of endoplasmic reticulum stress protein C/EBP homologous protein(CHOP)and caspase-3 in neurons,and quantitative PCR(qPCR)was used to detect changes in the inflammatory cytokines interleukin-1β(IL-1β)and IL-6 in brain tissue.In vitro cultured Neuro2a neurons were treated with heme or taurine sodium cholate(Tau,CHOP-specific inhibitor),and divided into control group,Hem group,Hem+Tau group,and Hem+Tau+Mar2 group.Western blotting was used to detect the levels of CHOP and caspase-3 expression in neurons;enzyme-linked immunosorbent assay was used to measure ROS levels.Results:Mar2 injection significantly improved neurological deficits,reduced brain edema,inhibited neuronal apoptosis,and a middle dose(10μg/kg/d)achieved the best therapeutic effect.Western blotting results showed that Mar2 could inhibit the expression of CHOP and caspase-3 in the cortex;immunofl

关 键 词：蛛网膜下腔出血 神经保护 Maresin2 C/EBP同源蛋白 

分 类 号：R741[医药卫生—神经病学与精神病学] R741.02[医药卫生—临床医学] R743.34