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背景:脑缺血再灌注后细胞周期蛋白的表达以其在凋亡机制中的作用为主,部分细胞凋亡是由于细胞增殖周期的异常调控所致。目的:研究脑缺血再灌注后细胞周期蛋白D1(cyclinD1)和周期蛋白依赖性蛋白激酶4(cyclindependentkinase,CDK4)基因表达及其与神经细胞凋亡的关系。设计:随机对照的实验研究。地点和材料:本实验在青岛大学医学院脑血管病研究所和山东省脑血管病防治重点实验室完成。成年健康雌性SD大鼠36只,体质量230～270g,清洁级,由中国科学院上海实验动物中心提供。将动物随机分为假手术组4只和实验组32只,实验组再进一步分为缺血1.5h再灌注2,6,12h,1,2,3,7和14d亚组,每组4只。方法:全部实验均由本文作者完成。具体方法:应用线栓法经左侧颈外-内动脉插线建立SD大鼠大脑中动脉阻塞再灌注模型,原位末端标记法检测脑缺血再灌注后神经细胞凋亡的变化,原位杂交技术检测cyclinD1mRNA和CDK4mRNA的表达。结果:脑缺血再灌注2h脑组织即开始出现凋亡神经细胞,并于1d和2d分别在皮质区和纹状体区达高峰(分别为72.80±4.66和96.75±4.37)。神经细胞cyclinD1mRNA和CDK4mRNA的表达分别于再灌注2h和6h开始逐渐增强,并于12h和1d分别在皮质区和纹状体区达高峰(皮质区分别为94.50±2.75和85.75±3.73,纹状体区分别为88.25?
BACKGROUND: The expression of cyclin in cerebral ischemia-reperfusion is mainly due to its role in the mechanism of apoptosis. Part of apoptosis is due to abnormal regulation of cell proliferation cycle. Objective: To investigate the relationship between cyclinD1 and CDK4 gene expression and neuronal apoptosis after cerebral ischemia-reperfusion. Design: Randomized controlled experimental study. Location and Materials: This experiment was performed at the Institute of Cerebrovascular Diseases, Medical College of Qingdao University and Key Laboratory of Prevention and Treatment of Cerebrovascular Diseases in Shandong Province. Thirty-six adult healthy female Sprague-Dawley rats weighing 230-270g were provided with the Shanghai Experimental Animal Center, Chinese Academy of Sciences. The animals were randomly divided into 4 sham-operation group and 32 experimental group. The experimental group was further divided into ischemia-reperfusion 1.5h reperfusion 2, 6, 12h, 1, 2, 3, 7 and 14d subgroups . Methods: All experiments were completed by the author. Specific methods: The middle cerebral artery occlusion and reperfusion model of SD rats was established by the method of thread plug on the left external carotid artery-internal artery, the change of neuronal apoptosis after cerebral ischemia-reperfusion was detected by in situ end labeling, Hybridization technique was used to detect the expression of cyclinD1mRNA and CDK4mRNA. Results: Apoptotic neurons began to appear in the brain 2h after cerebral ischemia-reperfusion, and peaked in cortex and striatum at 1d and 2d respectively (72.80 ± 4.66 and 96.75 ± 4.37, respectively). The expression of cyclinD1mRNA and CDK4mRNA in neurons gradually increased at 2h and 6h after reperfusion, respectively, and peaked in cortex and striatum at 12h and 1d respectively (cortical areas 94.50 ± 2.75 and 85.75 ± 3.73, District were 88.25?