【作者】 吕艳秋；

【导师】 石刚刚；

【作者基本信息】 汕头大学 ， 药理学， 2006， 硕士

【摘要】 2000年，Yan，S．F．等首先在《Nature Medicine》报道，Egr-1基因的激活在组织I／R损伤的发病机理中起主导作用。敲除Egr-1基因，能够明显减轻小鼠肺I／R损伤，保护肺功能。此后许多研究结果证实抑制Egr-1基因的转录与表达可以明显减少组织I／R时炎性细胞的浸润与活化，改善器官功能。本课题组以往研究结果证实碘化-N-正丁基氟哌啶醇（F₂）有保护心肌I／R损伤的作用，其机制可能与抑制心肌组织Egr-1的表达有关。为确认Egr-1在心肌I／R损伤发生发展过程中所起的作用，本实验采用大鼠心肌I／R模型，通过Egr-1 AS-ODN抑制大鼠心肌组织Egr-1蛋白的表达，从心肌组织炎症反应、脂质过氧化反应及心脏血流动力学、心肌酶释放等角度观察其对心肌I／R损伤的作用。 一．实验方法 1．给大鼠静脉注射5’-FITC标记的Egr-1 AS-ODN（4mg／kg），24h后，断头处死大鼠，取出心脏作常规冷冻切片。荧光显微镜400倍视野下，用488nm激发光照射，观察心肌组织对5’-FITC标记的Egr-1 AS-ODN的摄取，并拍摄照片。 2．大鼠随机分成5组：Sham（假手术）组、I／R（缺血再灌注）组、S（Egr-1正义寡核苷酸）组、Sc（错配寡核苷酸）组、AS（Egr-1反义寡核苷酸）组。缺血前24h静脉注射给药。24h后，制作大鼠心肌I／R模型：手术结扎大鼠冠状动脉左前降支（LAD）造成心肌缺血60min，然后解除结扎再灌注180min。并同步监测血流动力学各项指标：HR、SP、AP、LVSP、左心室内压变化速率最大值（±dp／dtmax）的变化。 实验结束后，取结扎线下缺血心肌组织，用Western-blot方法检测心肌组织Egr-1更多还原

【Abstract】 Yan, Shi. F. reported in 《Nature Medicine》that Egr-1 activation played a central and unifying role in the pathogenesis of I/R tissue injury in 2000. When the mice were knocked out Egr-1 gene, the lung I/R injury of the Egr-1-null mice was alleviated and lung function was improved. The succedent researches indicated that inhibition of Egr-1 gene decreased the activation of inflammatory cell and improved organ function. Results of the former researches in our department showed that N-n-butyl haloperidol iodide (F2) inhibited Egr-1 expression of myocardium of rats and alleviated myocardial I /R injury. In order to confirm the role of Egr-1 in myocardial I/R injury, we applied Egr-1 AS-ODN to inhibit Egr-1 protein of myocardium in rat’s myocardial I/R model. According to the results, we conclude that Egr-1 plays an important role in myocardial I/R injury.I. Methods1. Sprague-Dawley rat was administered 4 mg/kg of 5’-FITC-labelled AS-ODN for Egr-1 intravenously. After 24 h, the rat was executed and heart was taken out. Frozen sections were made and each slice was 5 urn in thickness. Then we observed the absorbtion of Egr-1 AS-ODN by fluorescence microscope and took pictures.2. Sprague-Dawley rats were randomly assigned to five groups: Sham group, I/R group, S group, Sc group and AS group. Each rat was administered intravenously 4 mg/kg of drug as described 24 h before ischemia. After 24 h, mycardial I/R model was produced by temporarily更多还原