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Am J Physiol Cell Physiol (October 20, 2004). doi:10.1152/ajpcell.00374.2004
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Submitted on July 30, 2004
Accepted on October 12, 2004

Expression of Constitutively Stable Hybrid Hypoxic Inducible Factor 1{alpha} Protects Cultured Rat Cardiomyocytes Against Simulated Ischemia/Reperfusion Injury

Taro Date1, Seibu Mochizuki2, Adam J Belanger1, Midori Yamakawa1, Zhengyu Luo1, Karen A Vincent1, Seng H Cheng1, Richard J Gregory1, and Canwen Jiang1*

1 Genzyme, Framingham, MA, USA
2 Department of Internal Medicine, Jikei University School of Medicine, Tokyo, Japan

* To whom correspondence should be addressed. E-mail: canwen.jiang{at}genzyme.com.

Preconditioning in cultured cardiomyocytes elevates the expression of several protective genes including Glut-4 and HSP70. Hypoxia-inducible factor 1 (HIF-1) is known to mediate the transcriptional activation of hypoxia responsive genes. In this study, we examined the effect of adenovirus-mediated expression of constitutively stable hybrid forms of HIF-1{alpha} on cardiomyocyte viability and gene expression. Cultured neonatal rat cardiomyocytes were subjected to simulated ischemia/reperfusion with or without pre-infection with recombinant adenoviral vectors (Ad2/HIF-1{alpha}/VP16 and Ad2/HIF-1{alpha}/NF{kappa}B). Cellular viability and mRNA levels of several cardioprotective genes were measured. We demonstrated that infection with Ad2/HIF-1{alpha}/VP16 and Ad2/HIF-1{alpha}/NF{kappa}B mimicked the up-regulation of the mRNA levels of VEGF, Glut-1, Glut-4, HSP70, and iNOS and the protection of cultured neonatal rat cardiomyocytes by late phase preconditioning against simulated ischemia/reperfusion. The same dose of a control viral vector expressing no transgene had no effect. Preconditioning also elevated HIF-1{alpha} protein levels. These results suggest that adenovirus-mediated expression of HIF-1{alpha}/VP16 or HIF-1{alpha}/NF{kappa}B, a constitutively stable hybrid transcriptional factor, protected cultured cardiomyocytes against simulated ischemia/reperfusion injury by inducing multiple protective genes.




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