中文摘要：

目的：构建急性心肌梗死（acute myocardial infarction，AMl）大鼠缺血心肌基因表达谱，对筛选出的目标基因进行功能研究。方法：结扎Wistar大鼠冠状动脉左前降支建立AMI模型，饲养7d处死，提取正常和缺血心肌总RNA，应用长标签基因表达系列分析（long serial analysis of geneexpression，LongSAGE）方法，构建AMI后大鼠缺血心肌基因差异表达谱，运用荧光定量PCR技术验证差异基因表达谱的可靠性，同时应用该技术定量两个功能基因的表达。生化法检测功能基因所表达的蛋白酶活性。结果：在建立的AMI后缺血心肌和正常心肌组织LongSAGE标签库中，共获得标签15966个，对照NCBI网站BLAST结果，发现新核苷酸序列标签7665个。与正常组比较，142个基因在AMI大鼠心肌组织中的表达差异有统计学意义（P〈0．05），这些差异基因主要与氧化磷酸化、三磷酸腺苷（ATP）合成、糖异生等能量代谢通路相关。荧光定量PCR的鉴定结果与LongSAGE差异表达结果基本一致。筛选能量代谢相关基因COXSa、ATP5e作为研究目标，荧光定量PCR结果提示2个基因在RNA水平表达下调；与之平行对应的功能蛋白酶的活性也相应降低。结论：AMI可引起多基因表达异常，能量代谢作用通路相关基因的异常表达可能是造成AMI心肌损伤的重要分子机制之一，干预COX5a和ATP5e等功能基因的表达可能是AMI能量代谢治疗的新靶点。

英文摘要：

Objective：To construct the differential genes expressed profile in the ischemic myocardium tissue reduced from acute myocardial infarction （ AMI）, and determine the biological functions of target genes. Methods： AMI model was generated by ligation of the left anterior descending coronary artery in Wistar rats. Total RNA was extracted from the normal and the ischemic heart tissues under the ligation point 7 days after the operation. Differential gene expression profiles of the two samples were constructed using Long Serial Analysis of Gene Expression （LongSAGE）. Real time fluorescence quantitative PCR was used to verify gene expression profile and to identify the expression of 2 functional genes. The activities of enzymes from functional genes were determined by histochemistry. Results. A total of 15 966 tags were screened from the normal and the ischemic LongSAGE maps. The similarities of the sequences were compared using the BLAST algebra in NCBI and 7 665 novel tags were found. In the ischemic tissue 142 genes were significantly changed compared with those in the normal tissue （P 〈 0. 05 ）. These differentially expressed genes represented the proteins which might play important roles in the pathways of oxidation and phosphorylation, ATP synthesis and glycolysis. The partial genes identified by LongSAGE were confirmed using real time fluorescence quantitative PCR. Two genes related to energy metabolism, COXSa and ATP5e, were screened and quantified. Expression of two functional genes down-regulated at their mRNA levels and the activities of correlative functional enzymes decreased compared with those in the normal tissue. Conclusion： AM! causes a series of changes in gene expression, in which the abnor- mal expression of genes related to energy metabolism could be one of the molecular mechanisms of AMI. The intervention of the expressions of COXSa and ATPSe may be a new target for AMI therapy.