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Acta Physiologica 2007; Volume 190, Supplement 656
The Scandinavian Physiological Society's Annual Meeting
8/10/2007-8/12/2007
Oslo, Norway
TRANSFECTION OF POTENTIAL CARDIOPROTECTIVE FACTORS INTO A CARDIOMYOCYTE MODEL TO INVESTIGATE SURVIVAL
Abstract number: P07
Sagave1 J, Czibik1 G, Martinov1 V, Valen1 G
1Institute of Basic Medical Science, Department of Physiology, University of Oslo, OSLO, Norway
Gene delivery of the transcription factor hypoxia- inducible factor 1-alpha (HIF-1) to the skeletal muscle was found to improve cardiac tolerance to induced ischemia. It also upregulated potential cardioprotective genes, such as adrenomedullin, heme oxygenase-1, insulin- like growth factor 2 and platelet derived growth factor-BB (PDGF-BB) in the treated quadriceps muscle. The aim of this study was to establish a model of induced cell death in immortalized murine cardiomyocyte HL-1 cells and to transfect these genes to test their cell-protective properties. Transfection was performed by pcDNA3.1(+) (empty vector) or the cloned target vectors by LIPOFECTAMINE 2000 (Invitrogen, Norway), which transfected > 65 % of the cells (enhanced green fluorescent protein (EGFP) reporter). Transfection efficacy was evaluated also at RNA level by real-time PCR or at protein level by immunhistochemistry, and demonstrated largely increased expression of HIF-1/ PDGF-BB. 44 h later cells were stimulated with hydrogen peroxide (H2O2) 300 mM for 4 h which caused death of approximately 3040 % of the cells (trypan blue). Transfection with HIF-1 or PDGF-BB resulted in a ~30 % reduction of dead HL-1 cells after stimulation with H2O2 (trypan blue), compared to the cells transfected with only empty vector. In conclusion, transfection of HIF-1 and PDGF-BB improved survival of HL-1 cardiomyocytes after H2O2-treatment. The other HIF-1 regulated genes are currently investigated.
To cite this abstract, please use the following information:
Acta Physiologica 2007; Volume 190, Supplement 656 :P07