脑缺血再灌注后神经细胞凋亡与碱性成纤维细胞生长因子及其受体表达的关系(英文)  

Neuronal apoptosis associated with basic fibroblast growth factor and its receptor following cerebral ischemia reperfusion

作　　者：丁晓洁[1] 孙峰[1] 王超[1] 郭云良[2] 

机构地区：[1]青岛大学医学院第二附属医院,山东省青岛市266042 [2]青岛大学医学院附属医院脑血管病研究所,山东省青岛市266003

出　　处：《中国组织工程研究与临床康复》2007年第14期2776-2779,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：山东省自然科学基金资助项目(Y2004C04)~~

摘　　要：背景:脑受到损害可诱导脑内碱性成纤维细胞生长因子表达增加,而成纤维细胞生长因子受体在细胞的增殖、分化、血管生成、骨骼形成等进程中起着重要作用。目的:观察大鼠脑缺血再灌注后碱性成纤维细胞生长因子及成纤维细胞生长因子受体1表达对神经细胞凋亡的影响。设计:随机分组设计、动物实验。单位:青岛大学医学院附属医院脑血管病研究所。材料:选用28只成年健康雄性Wistar大鼠,体质量220～260g,清洁级,由山东大学实验动物中心提供。兔抗大鼠碱性成纤维细胞生长因子和成纤维细胞生长因子受体1单克隆抗体由武汉博士德生物工程有限公司提供。方法:实验于2005-05/12在山东省脑病防治重点实验室完成。①摸球法将大鼠随机分为实验组(n=24)和假手术组(n=4)。应用线栓法经左侧颈外-内动脉插线建立大脑中动脉闭塞再灌注模型,假手术组除不插线外,其余步骤同实验组。实验组大鼠于脑缺血1h再灌注6h、12h、1d、3d、7d、14d各时间点取4只大鼠进行标本取材,假手术组于术后24h取材。大鼠麻醉后断头完整取脑,切取视交叉后方约5mm的脑组织,连续冠状切片备用。②脑组织经苏木精-伊红染色,显微镜下观察大鼠神经细胞形态。③TUNEL法:细胞核出现棕黄色颗粒者为阳性着色,即凋亡细胞。高倍镜下在皮质区和纹状体区随机各取4个视野计数阳性细胞。④标本切片经兔抗大鼠bFGF和FGFR-1单克隆抗体染色,高倍镜下在皮质区和纹状体区随机各取4个视野计数阳性细胞。主要观察指标:各组大鼠神经细胞凋亡情况及碱性成纤维细胞生长因子和成纤维细胞生长因子受体1表达情况。结果:纳入28只大鼠全部进入结果分析。①实验组大鼠脑缺血损伤区神经细胞数量明显减少,部分细胞出现核固缩、不规则,染色加深呈紫蓝色,核仁消失,并有许多散在的细胞碎片。②假手术�BACKGROUND： Brain injury can induce the increased expression of basic fibroblast growth factor （bFGF） in brain, whereas FGFR is a very important player in the cell proliferation and differentiation, angiogenesis, skeletogeny, etc. OBJECTIVE： To observe the effect of bFGF and its receptor on neuronal apoptosis following cerebra ischemia/reperfusion injury in rats. DESIGN： A randomized grouping design and animal experiment. SETTING： Institute of Cerebrovascular Disease, Affiliated Hospital of Qingdao University Medical College. MATERIALS： Twenty-eight healthy adult Wistar rats of clean degree, weighing 220-260 g, were provided by the experimental animal center of Shandong University. Rabbit-anti-rat bFGF and fibroblast growth factor receptor-1 （FGFR-1） monoclonal antibodies were provided by Wuhan Boster Biological Technology, Co.,Ltd. METHODS： The experiment was carried out in Shandong Key Laboratory for Prevention and Treatment of Brain diseases. ① The rats were randomly divided into experimental group （n =24） and sham-operated group （n =4）. Models of middle cerebral artery occlusion/reperfusion （MCAO/R） were established by thread occlusion via left external-internal carotid arteries, and 4 rats in the experimental group were sampled at 1-hour ischemia/6, 12-hour, 1, 3, 7 and 14-day reperfusion respectively. The rats in the sham-operated group were given the same treatment without inserting thread. After anesthesia, the brain was removed completely by cutting head, then the brain tissue at about 5 mm posterior to optic chiasma was cut down, then serial coronal sections （5 μm） were prepared. ② The brain tissues were stained with ematoxylin-eosin （HE）, and the forms of neurons were observed under microscope. ③ TdT-mediated dUTP-biotin nick end labeling （TUNEL） method： there were buffy granules in nucleus which was positively stained （apoptosis）. Four fields were randomly selected from cortex and striatum to count positive cells under high-power micros

关 键 词：脑缺血 再灌注 成纤维细胞生长因子 碱性 受体 生长因子 

分 类 号：R743[医药卫生—神经病学与精神病学]