局部脑缺血再灌注损伤对小鼠TMEM166基因和神经细胞自噬的影响  

Effect of focal cerebral ischemia/reperfusion injury on TMEM166 and neuron autophagy

作　　者：王瑜[1] 王天龙[1] 赵磊[1] 李丽[1] WANG Yu;WANG Tianlong;ZHAO Lei;LI Li(Department of Anesthesiology,Xuanwu Hospital,Capital Medical University,Beijing 100053,China)

机构地区：[1]首都医科大学宣武医院麻醉科,北京100053

出　　处：《中国比较医学杂志》2018年第7期28-32,共5页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金项目(编号:81500996);中国博士后科学基金资助项目(编号:2016T90114);北京市医院管理局"登峰计划"专项经费资助(编号:DFL20150802);215高层次卫生技术人才学术骨干项目(编号:008-0093)

摘　　要：目的观察局部脑缺血再灌注损伤对小鼠TMEM166基因和神经细胞自噬的影响。方法C57/BL6J小鼠50只,线栓法制作大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)局部脑缺血模型,随机分为再灌注6,12,24,48 h组。通过免疫组化染色观察缺血侧大脑皮层及基底核TMEM166、LC3-II的阳性细胞数量变化,Western blot检测不同再灌注时间TMEM166、LC3-II蛋白水平的表达。结果缺血侧TMEM166随再灌注时间延长而增加,24 h达到高峰;再灌注6 h后LC3-II表达出现,同样随再灌注时间而增加,24 h达到高峰。缺血侧自噬细胞数量的变化趋势与TMEM166基本一致,对侧大脑半球上述指标无明显变化。结论小鼠局部脑缺血再灌注损伤可以诱导TMEM166的表达,通过激活LC3-II引起神经细胞自噬。Objective To investigate the effect of focal cerebral ischemia/reperfusion injury on TMEM166 and neuron autophagy.Methods In total,fifty C57/BL6J mice were used to establish the focal cerebral ischemia model through middle cerebral artery occlusion(MCAO)by endovascular suture and were randomly divided into four groups according to different reperfusion time points:6,12,24 and 48 h.Changes in the expression of TMEM166 and the number of LC3-II positive cells in cerebral cortex and basal ganglia of the ischemic hemisphere of the mice were measured by immunochemistry.Protein levels of TMEM166 and LC3-II were detected by Western blotting.Results TMEM166 was induced in mice after cerebral ischemia/reperfusion injury,peaking at 24 h after injury in the ischemic hemisphere.LC3-II began to increase at 6 h following MCAO,increasing over time to reach a highest level at 24 h after ischemic insult.The changing pattern of TMEM166 was consistent with changes in LC3-II positive cell numbers.No changes of TMEM166 and LC3-II were found in the contralateral hemisphere cerebral cortex and basal ganglia.Conclusions TMEM166 is induced after MCAO,and can lead to neuron autophagy by activating LC3-II.

关 键 词：TMEM166基因 脑缺血再灌注 自噬 小鼠 

分 类 号：R-33[医药卫生]