Heterozygous disruption of Flk-1 receptor leads to myocardial ischaemia reperfusion injury in mice: Application of affymetrix gene chip analysis

M. Thirunavukkarasu, S. Addya, B. Juhasz, R. Pant, L. Zhan, S. Surrey, G. Maulik, V. P. Menon, N. Maulik

Research output: Contribution to journalArticle

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Abstract

This study addresses an important clinical issue by identifying potential candidates of vascular endothelial growth factor (VEGF) signalling through the Flk-1 receptor that trigger cardioprotective signals under ischaemic stress. Isolated working mouse hearts of both wild-type (WT) and Flk-1+/- were subjected to global ischaemia (I) for 30 min. followed by 2 hrs of reperfusion (R). Flk-1+/- myocardium displayed almost 50% reduction in Flk-1 mRNA as examined by quantitative real-time RT-PCR at the baseline level. Flk-1+/- mouse hearts displayed reduction in left ventricular functional recovery throughout reperfusion (dp/dt 605 versus 884), after 2 hrs (P <0.05). Coronary (1.9 versus 2.4 ml) and aortic flow (AF) (0.16 versus 1.2 ml) were reduced in Flk-1+/- after 2 hrs of reperfusion. In addition, increased infarct size (38.4% versus 28.41%, P <0.05) and apoptotic cardiomyocytes (495 versus 213) were observed in Flk-1+/- knockout (KO) mice. We also examined whether ischaemic preconditioning (PC), a novel method to induce cardioprotection against ischaemia reperfusion injury, through stimulating the VEGF signalling pathway might function in Flk-1 +/- mice. We found that knocking down Flk-1 resulted in significant reduction in the cardioprotective effect by PC compared to WT. Affymetrix gene chip analysis demonstrated down-regulation of important genes after IR and preconditioning followed by ischaemia reperfusion in Flk-1+/- mice compared to WT. To get insight into the underlying molecular pathways involved in ischaemic PC, we determined the distinct and overlapping biological processes using Ingenuity pathway analysis tool. Independent evidence at the mRNA level supporting the Affymetrix results were validated using real-time RT-PCR for selected down-regulated genes, which are thought to play important roles in cardioprotection after ischaemic insult. In summary, our data indicated for the first time that ischaemic PC modifies genomic responses in heterozygous VEGFR-2/Flk-1 KO mice and abolishes its cardioprotective effect on ischaemic myocardium.

Original languageEnglish
Pages (from-to)1284-1302
Number of pages19
JournalJournal of Cellular and Molecular Medicine
Volume12
Issue number4
DOIs
Publication statusPublished - Aug 2008

Fingerprint

Myocardial Reperfusion Injury
Oligonucleotide Array Sequence Analysis
Reperfusion Injury
Ischemic Preconditioning
Reperfusion
Myocardial Ischemia
Genes
Vascular Endothelial Growth Factor A
Knockout Mice
Real-Time Polymerase Chain Reaction
Vascular Endothelial Growth Factor Receptor-2
Myocardium
Messenger RNA
Ischemia
Biological Phenomena
Cardiac Myocytes
Recovery
Down-Regulation

Keywords

  • Affymetrix gene chip
  • Flk-1
  • Gene expression
  • Ischaemia
  • Myocardium
  • Reperfusion

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Heterozygous disruption of Flk-1 receptor leads to myocardial ischaemia reperfusion injury in mice : Application of affymetrix gene chip analysis. / Thirunavukkarasu, M.; Addya, S.; Juhasz, B.; Pant, R.; Zhan, L.; Surrey, S.; Maulik, G.; Menon, V. P.; Maulik, N.

In: Journal of Cellular and Molecular Medicine, Vol. 12, No. 4, 08.2008, p. 1284-1302.

Research output: Contribution to journalArticle

Thirunavukkarasu, M. ; Addya, S. ; Juhasz, B. ; Pant, R. ; Zhan, L. ; Surrey, S. ; Maulik, G. ; Menon, V. P. ; Maulik, N. / Heterozygous disruption of Flk-1 receptor leads to myocardial ischaemia reperfusion injury in mice : Application of affymetrix gene chip analysis. In: Journal of Cellular and Molecular Medicine. 2008 ; Vol. 12, No. 4. pp. 1284-1302.
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