短暂脑缺血再灌注后突触蛋白I表达与细胞凋亡  

Relationship between expression of synapsin I and apoptosis after transient ischemia followed by reperfusion

作　　者：梁燕玲[1] 张苏明[1] 许康[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院神经内科,湖北省武汉市430030

出　　处：《中国临床康复》2005年第37期54-56,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金面上项目(30070825)~~

摘　　要：目的:观察短暂脑缺血再灌注对与神经元的早期发育和再生相关的突触蛋白I及神经细胞凋亡的影响,进一步了解神经系统的可塑性。方法:实验于2003年于同济医院神经内科实验室进行。①分组:取75只Wister大鼠随机分为模型组(n=51)、假手术组(n=18)和正常对照组(n=6)3组。假手术组18只和模型组18只分为再灌注1,3,6,12,24和72h6个亚组,进行突触蛋白I检测;正常对照组6只和模型组剩余的33只大鼠(分为再灌注1,3,6,12,24,48和72h7个亚组)分别进行TUNEL阳性细胞检测、形态学观察和流式细胞仪检测,需获取数据时至少检测2只大鼠。②造模:模型组采用线栓法建立大鼠大脑中动脉闭塞模型,缺血10min后再灌注;假手术组不插入线栓,其余步骤同模型组;正常对照组不干预。③观察指标:于相应时间点麻醉状态下处死取脑,应用免疫组织化学的方法观察缺血侧额顶叶皮质突触蛋白I表达的动态变化,同时采用荧光显微镜、流式细胞仪和脱氧核苷酸末端转移酶介导的缺口末端标记法检测细胞凋亡情况,分析两者之间的关系。结果:经补充后72只大鼠进入结果分析。①缺血侧额顶叶皮质突触蛋白I的表达:模型组再灌注12h内无明显变化,再灌注24h低于假手术组(0.199±0.006,0.238±0.008,P<0.01),至72h恢复至假手术组水平。②细胞凋亡率:模型组再灌注24,48和72h均高于正常对照组[(12.57±9.83)%,(13.56±2.28)%,(16.68±0.66)%,(4.65±0.03)%,P<0.05]。③TUNEL阳性细胞率:正常对照组和模型组再灌注0～24h均未见阳性细胞,再灌注48和72h分别为(47.50±3.85)%和(62.66±13.06)%。结论:短暂脑缺血再灌注后存在着突触蛋白I表达的短暂降低,且与凋亡细胞的出现在时间上非常吻合,提示短暂脑缺血再灌注后存在失神经及其后的神经再获现象,这可能与DNA的损伤和修复有关。AIM： To observe the effects of transient ischemia followed by repufusion on the synapsin Ⅰ, which is associated with the early development and regeneration of neuron, and apoptosis of nerve ceils, and further investigate the plasticity of nervous system. METHODS： The experiment was carried out in the laboratory of Department of Neurology, Tongji Hospital in 2003. ① Grouping： Seventy-five Wistar rats were randomly divided into model group （n=51）, sham-operated group （n=18） and normal control group （n=6）. The 18 rats in the sham-operated group and 18 rats in the model group were subdivided in to 1, 3, 6, 12, 24 and 72-hour reperfusion groups respectively for the detection of synapsin Ⅰ. The 6 rats in the normal control group and the rest 33 rats in the model group were subdivided in to 1, 3, 6, 12, 24 and 72-hour reperfusion groups respectively, and the positive ceils were detected with terminal deoxynucleotidyl transferase mediated d-UTP nick-end labeling （TUNEL）, morphological observation, flow eytometer detection were also performed, at least 2 rats were detected for obtaining the data. ② Model establishment： Rats in the model group were made into models of middle cerebral artery occlusion （MCAO） by thread embolism method, and then treated with ischemia for 10 minutes followed by reperfusion. Rats in the sham-operated group were not be inserted the thread, others were the same as those in the model group. No intervention was given to the rats in the nonrkal control group. ③ Observed indexes： The rats were killed to remove brain at each corresponding time point under anesthesia, and the dynamic changes of the synapsin I expression of in the frontal-parietal cortex of the ischemic side were observed by means of immunohistochemistry. At the same time, apoptosis was measured by fluoroscope, flow cytometer and TUNEL. The relationship between them was analyzed at last. RESULTS： Seventy-two rats were bought into the analysis of results after completion. ① The synapsin Ⅰ exp

关 键 词：脑缺血 再灌注 突触蛋白类 细胞凋亡 

分 类 号：R743[医药卫生—神经病学与精神病学]