实验性血管性痴呆小鼠中枢神经系统细胞凋亡与迟发性神经元坏死  

Cell apoptosis and delayed neuronal death in central nervous system of experimental mice with vascular dementia

作　　者：李巍[1] 姜立刚[2] 徐忠信[3] 饶明俐[4] 

机构地区：[1]北华大学第一临床医院神经内科,吉林省吉林市132001 [2]北华大学附属医院神经内科,硕士研究生吉林省吉林市132001 [3]吉林大学中日联谊医院神经内科,吉林省长春市130021 [4]吉林大学第一医院神经内科,吉林省长春市130021

出　　处：《中国临床康复》2005年第28期133-135,F0003,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:探讨实验性血管性痴呆小鼠中枢神经系统细胞凋亡与迟发性神经元坏死的关系。方法:实验于1998-01/1999-07在白求恩医科大学实验动物中心完成。选用6～7周龄雄性昆明小鼠40只,随机分为假手术组和缺血再灌流组;假手术组5只作为对照,缺血再灌流组根据再灌注提取标本的时间不同并分7组,即术后1h,1,3,7,14,28,42d组,每小组5只。制备动物模型,选用跳台,避暗反应等行为学检测确定。评价采用卒中指数评分和神经病学症状评分标准。制作病理切片,应用光镜,电镜,流式细胞仪,原位末端标记等方法检测神经细胞凋亡在不同时间缺血再灌注时与迟发性神经元坏死发生、发展关系。结果:在整个实验过程中无动物死亡,全部进入结果分析。①动物模型卒中指数及神经病学指数评价:缺血再灌注1d时卒中指数及神经病学指数与对照组比较差异显著(3.5±0.53,0,P<0.05),(4.6±0.54,0,P<0.05)。②各组病理形态学改变:重复缺血再灌注24h神经细胞间质水肿,核浓染固缩,顶树突延长。再灌注3d,皮质少量小胶质细胞增生,海马可见颗粒细胞变性呈空泡样。再灌注7d,皮质胶质细胞增生聚集成堆,海马CA1区锥体细胞变性,部分坏死。再灌注28d,皮质神经细胞大量缺失代之胶质细胞,海马CA1、CA4区锥体细胞大量缺失、变性、坏死。电镜观察可见细胞核基质肿胀,破坏。排列紊乱,并有空泡形成。粗面内质网扩张,小胶质细胞核染色质凝聚,可见变形。③额叶、海马区细胞凋亡变化:重复缺血再灌注3d流式细胞仪检测额叶皮质、海马神经细胞凋亡百分率最高。随再灌注时间的延长,海马神经细胞凋亡数逐渐下降至42d最低点。各不同时间点再灌注细胞凋亡数百分比与对照组比较差异显著性(P<0.05)。原位末端标记法正常阳性对照组可见少量散在阳性细胞,缺血再灌注1h后原位末端标记法染色阳性细胞增�AIM： To investigate the relativity of central nerve cell apoptosis and lateonset necrosis of neurons in experimental mice with vascular dementia. METHODS： The experiment was conducted in the Center for Experimental Animals, Bethune Medical University from January 1998 to July 1999. Forty Kunming male mice at the age of 6 to 7 weeks were randomly divided into sham operation group（n=5） and ischemia-reperfusion （IR） group （n=35）, and the latter group were equally divided into seven groups according to the sampling time hy reperfusion： 1 post-IR hour, 1, 3, 7,14, 28 and 42 post-IR groups. The mice were modeled, and then were evaluated according to the criteria of stroke index and neurological symptom scoring. The pathological sections were made to detect the relationship between the nerve cell apoptosis at different IR time and the episode and development of late-onset necrosis of neurons by means of light microscopy, electron microscopy, flow cytometry, in situ end-labeling, etc. RESULTS： All the mice were analyzed in the result without loss. ① Evaluation of stroke index and neurological index in the animal models： Both stroke index and neurological index were significantly different between the 1 post-lR day group and the sham operation group（3.5±0.53, 0 vs 4.6±0.54, 0, both P 〈 0.05）.②Pathological and morphological changes in all the groups： At 24 hours after repeated IR, interstitial edema of nerve cells was found, nuclei were stained deeply and pyknotic, and the dendrite of the top was prolonged. At 3 days after reperfusion, a small quantity of microglia cells in cortex were proliferative, and granular ceils in hippocampus were degenerative in vacuole-like shape. At 7 days after reperfusion, microglia cells in cortex proliferated and gathered into piles, and pyramidal cells in hippocampal CA1 area were degenerative, some of which were necrotic. At 28 days after reperfusion, lots of microglia cells in cortex disappeared, and substituted by glial cells; The pyramidal cells in hi

关 键 词：细胞凋亡 痴呆 血管性 小胶质神经细胞 

分 类 号：R749[医药卫生—神经病学与精神病学]