MEK-ERK介导NMDA受体激活后的脑出血损伤的机制  

Mechanism of MEK-ERK mediating the intracerebral hemorrhage after NMDA receptor activation

作　　者：王光绿[1] 邓峰[1] 莫贻敏 简俊红[1] WANG Guanglü;DENG Feng;MO Yimin;JIAN Junhong(Department of Neurosuregy, Beihai People’ s Hospital, Beihai 536000, China)

机构地区：[1]广西北海市人民医院神经外科,北海536000

出　　处：《山西医科大学学报》2019年第5期590-594,共5页Journal of Shanxi Medical University

基　　金：北海市科技局科研项目(北科合201203050)

摘　　要：目的探讨MEK-ERK介导NMDA受体激活后的脑出血损伤的机制。方法 4周龄SPF级雄性BALB/c近交系小鼠30只,随机分为实验组(n=15)和对照组(n=15),实验组将血液缓慢均匀地注入大脑,构建小鼠脑出血模型;对照组注射生理盐水。Menzies评分法观察两组小鼠神经功能缺损情况,TUNEL染色法计算凋亡指数来评价脑出血损伤后海马神经元细胞的凋亡情况,免疫印迹法检测海马组织MEK、ERK、NFATc4、Rho A和PKCα蛋白的表达水平,OHG-300型氢清除法组织血流仪测定局部脑血流量。结果 Menzies评分法观察小鼠神经功能缺损情况,结果显示实验组小鼠的神经功能缺损评分显著高于对照组,在出血6 h神经损害最显著,差异有统计学意义(P <0. 05)。在脑出血的小鼠中MEK-ERK通路相关蛋白MEK、ERK以及钙调神经磷酸酶NFATc4表达量显著上调(P <0. 05),NMDA受体调控蛋白PKCα、Rho A的表达较正常对照显著上升(P <0. 05),TUNEL法检测实验组脑细胞凋亡数显著多于对照组(P <0. 05),实验组小鼠大脑局部血流量显著下降(P<0. 05)。结论在脑出血的小鼠中MEK-ERK通路相关蛋白MEK、ERK以及钙调神经磷酸酶NFATc4表达水平显著上调,诱发神经功能缺损。Objective To explore the mechanism of MEK-ERK mediating brain hemorrhage injury after activation of NMDA receptor.Methods Thirty SPF male BALB/c inbred mice aged 4 weeks were randomly divided into experimental group( n = 15) and control group( n = 15). The blood was slowly and evenly injected into the brain to establish a model of cerebral hemorrhage in mice in experimental group,while the mice were injected with normal saline in control group. Menzies scoring method was used to observe the neurological deficits in the two groups. TUNEL staining was used to calculate the apoptotic index for evaluating the apoptotic status of hippocampal neurons after cerebral hemorrhage injury. Immunoblotting was used to detect the expression levels of MEK,ERK,NFATc4,Rho A and PKC alpha proteins in hippocampus. OHG-300 tissue hemodynamic instrument was used to measure the regional cerebral blood flow. Results Menzies scoring results showed that the neurological deficit score in experimental group was significantly higher than that in control group( P < 0. 05),especially at 6 h after bleeding. The expression of MEK,ERK and calcineurin NFATc4 related proteins in MEK-ERK pathway were significantly up-regulated in mice with cerebral hemorrhage( P < 0. 05). The expression of NMDA receptor regulatory proteins PKC alpha and Rho A was significantly higher in experimental group than that in control group( P < 0. 05).The number of apoptotic brain cells in experimental group was significantly higher than that in control group by TUNEL( P < 0. 05).The regional cerebral blood flow of mice in experimental group was significantly lower than that in control group( P < 0. 05). Conclusion The expression levels of MEK,ERK and calcineurin NFATc4 related proteins in MEK-ERK pathway are significantly up-regulated in mice with intracerebral hemorrhage,which may induce neurological deficits.

关 键 词：脑出血 MEK-ERK通路 NFAT 细胞凋亡 NMDA 

分 类 号：R743.34[医药卫生—神经病学与精神病学]