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Physiol. Genomics (March 22, 2006). doi:10.1152/physiolgenomics.00013.2006
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Submitted on January 27, 2006
Accepted on March 3, 2006

Transcriptome analysis of the ischemia-reperfused remodeling myocardium: temporal changes in inflammation and extracellular matrix

Sashwati Roy1, Savita Khanna1, Donald E Kuhn1, Cameron Rink1, Willis T Williams1, Jay L Zweier1, and Chandan K. Sen1*

1 Davis Heart and Lung Research Institute, The Ohio State University Medical Center, Columbus, Ohio, United States

* To whom correspondence should be addressed. E-mail: chandan.sen{at}osumc.edu.

cDNA microarray analysis was performed to screen 15,000 genes and 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 d or 7 d only) or on a more sustained basis (2d and 7d) were derived. The candidate genes represented three major functional categories- extracellular matrix, apoptosis and inflammation. To expand on the findings from microarray studies which dealt with the two above-mentioned timepoints, tissues collected from days 0, 0.25, 2, 3, 5 and 7 after reperfusion were examined. Acute myocardial infarction resulted in up-regulation of IL-6 and IL-18. Genes encoding extracellular matrix proteins such as types I and III collagen were up-regulated 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 to 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 (LMPC) technology was applied to harvest infarct and adjacent non-infarct control tissue from microscopically defined region in the rat myocardium. Taken together, this work presents first evidence gained from the use of DNA microarrays to understand the molecular mechanisms implicated in the early and late inflammatory phases of the ischemia-reperfused heart.




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