Roy Sashwati, Khanna Savita, Kuhn Donald E, Rink Cameron, Williams Willis T, Zweier Jay L, Sen Chandan K
Department of Surgery, Davis Heart and Lung Research Institute, The Ohio State University Medical Center, Columbus, Ohio, USA.
Physiol Genomics. 2006 May 16;25(3):364-74. doi: 10.1152/physiolgenomics.00013.2006. Epub 2006 Mar 22.
cDNA microarray analysis was performed to screen 15,000 genes and expressed sequence tags (ESTs) to identify changes in the ischemia-reperfused (I-R) rat myocardial transcriptome in the early (day 2) and late (day 7) inflammatory phases of acute myocardial infarction. Lists of candidate genes that were affected by I-R transiently (2 or 7 days only) or on a more sustained basis (2 and 7 days) were derived. The candidate genes represented three major functional categories: extracellular matrix, apoptosis, and inflammation. To expand on the findings from microarray studies that dealt with the two above-mentioned time points, tissues collected from days 0, 0.25, 2, 3, 5, and 7 after reperfusion were examined. Acute myocardial infarction resulted in upregulation of IL-6 and IL-18. Genes encoding extracellular matrix proteins such as types I and III collagen were upregulated in day 2, and that response progressively grew stronger until day 7 after I-R. Comparable response kinetics was exhibited by the candidate genes of the apoptosis category. Caspases-2, -3, and -8 were induced in response to acute infarction. Compared with the myocardial tissue from the sham-operated rats, tissue collected from the infarct region stained heavily positive for the presence of active caspase-3. Laser microdissection and pressure catapulting technology was applied to harvest infarct and adjacent noninfarct control tissue from a microscopically defined region in the rat myocardium. Taken together, this work presents the first evidence gained from the use of DNA microarrays to understand the molecular mechanisms implicated in the early and late inflammatory phases of the I-R heart.
进行cDNA微阵列分析以筛选15000个基因和表达序列标签（EST），以确定急性心肌梗死早期（第2天）和晚期（第7天）炎症阶段缺血再灌注（I-R）大鼠心肌转录组的变化。得出了受I-R短暂影响（仅2天或7天）或持续影响（2天和7天）的候选基因列表。候选基因代表三个主要功能类别：细胞外基质、细胞凋亡和炎症。为了扩展处理上述两个时间点的微阵列研究结果，对再灌注后第0、0.25、2、3、5和7天收集的组织进行了检查。急性心肌梗死导致IL-6和IL-18上调。编码细胞外基质蛋白（如I型和III型胶原蛋白）的基因在第2天上调，并且这种反应在I-R后直到第7天逐渐增强。细胞凋亡类别的候选基因表现出类似的反应动力学。Caspases-2、-3和-8因急性梗死而被诱导。与假手术大鼠的心肌组织相比，从梗死区域收集的组织中活性caspase-3染色呈强阳性。应用激光显微切割和压力弹射技术从大鼠心肌中显微镜定义的区域收获梗死和相邻的非梗死对照组织。综上所述，这项工作提供了从使用DNA微阵列获得的首个证据，以了解I-R心脏早期和晚期炎症阶段涉及的分子机制。
