NMDA受体介导脑缺血/再灌注诱导GluR6巯基亚硝基化的研究  

NMDA Receptor Involved in Cerebral Ischemia-Reperfusion Induced GluR6S-nitrosylation

作　　者：王梅 戚大石 胡晓彤 刘亚萍 张芳 

机构地区：[1]徐州医科大学形态学实验教学中心江苏省脑病重点实验室,江苏徐州221002

出　　处：《现代生物医学进展》2016年第32期6235-6238,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金青年基金项目(81500977);江苏省高校自然科学研究面上项目(14KJB180022);江苏省脑病生物信息重点实验室开放研究课题(1505);江苏省普通高校自然科学研究项目(13KJD310003);徐州市科技计划项目(KC14SH076)

摘　　要：目的:研究脑缺血再灌注以及联合给予脑缺血和NMDA(N-甲基-D-天冬氨酸)受体抑制剂MK801对大鼠海马CA1区Glu R6巯基亚硝基化以及海马CA1区锥体细胞凋亡的影响。方法:采用四动脉结扎法构建大鼠全脑缺血再灌注模型,给予SD大鼠腹腔注射NMDA受体特异性抑制剂MK801(3 mg/kg)。主要运用'生物素转化法'(Biotin-Switch method)、SDS-PAGE、免疫印迹、焦油紫染色等方法对Glu R6的巯基亚硝基化(S-亚硝基化)、蛋白表达水平以及海马CA1区锥体细胞的凋亡水平进行研究。结果:脑缺血/再灌注显著促进Glu R6的巯基亚硝基化以及海马CA1区锥体细胞的凋亡,给予NMDA受体特异性抑制剂MK801能够显著抑制脑缺血/复灌诱导增加的Glu R6的S-亚硝基化以及海马CA1区锥体细胞的凋亡。结论:脑缺血/再灌注早期NMDA受体介导了Glu R6的巯基亚硝基化以及海马CA1区锥体细胞的凋亡,从而为临床治疗缺血再灌注脑损伤提供了理论依据。Objective： The objective of this study is primarily to determine whether NMDA recepter affects Glu R6 S-nitrosylation and the survival of CA1 subfield during the early stages of ischemia-reperfusion. Methods： Transient cerebral ischemia was induced by a four-vessel occluded（4-VO） method. Sprague-Dawley rats were treated intraperitoneally with MK801（3 mg/kg, an selective inhibitor of NMDA recepter）. We perform this study mainly by biotin switch assay, SDS-PAGE and western bloting to exam the S-nitrosylation and expression of Glu R6. Cresyl violet staining was performed to examine the survival and apoptosis neurons in the pyramidal cell layer of the hippocamal CA1 subfield. Results： Here, we showed that S-nitrosylation of Glu R6 and the apoptosis of CA1 pyramidal neuron was dramatically induced by cerebral ischemia-reperfusion,and that administration of MK801 observably diminished the increased S-nitrosylation of Glu R6 and the apoptosis of CA1 pyramidal neuron. Conclusions： These data suggest that NMDA recepter facilitates Glu R6 S-nitrosylation and the apoptosis of CA1 pyramidal neuron in the early stages of cerebral ischemia-reperfusion. In contrast,MK801 antagonizes the above action of cerebral ischemia-reperfusion.Thus, our results provide that NMDA recepter regulate Glu R6 by S-nitrosylation and affect the CA1 pyramidal neuron during the early stages of ischemia-reperfusion, which can be a new approach for stroke therapy.

关 键 词：脑缺血再灌注 NMDA受体 S-亚硝基化 谷氨酸受体6 

分 类 号：Q95-3[生物学—动物学] R743[医药卫生—神经病学与精神病学]