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1.
Exp. mol. med ; Exp. mol. med;: 674-683, 2012.
Article de Anglais | WPRIM | ID: wpr-149761

RÉSUMÉ

Relative deficiency in production of glycoprotein hormone erythropoietin (Epo) is a major cause of renal anemia. This study planned to investigate whether the hypoxia-regulated system of Epo expression, constructed by fusing Epo gene to the chimeric phosphoglycerate kinase (PGK) hypoxia response elements (HRE) in combination with cytomegalovirus immediate-early (CMV IE) basal gene promoter and delivered by plasmid intramuscular injection, might provide a long-term physiologically regulated Epo secretion expression to correct the anemia in adenine-induced uremic rats. Plasmid vectors (pHRE-Epo) were synthesized by fusing human Epo cDNA to the HRE/CMV promoter. Hypoxia-inducible activity of this promoter was evaluated first in vitro and then in vivo in healthy and uremic rats (n = 30 per group). The vectors (pCMV-Epo) in which Epo expression was directed by a constitutive CMV gene promoter served as control. ANOVA and Student's t-test were used to analyze between-group differences. A high-level expression of Epo was induced by hypoxia in vitro and in vivo. Though both pHRE-Epo and pCMV-Epo corrected anemia, the hematocrit of the pCMV-Epo-treated rats exceeded the normal (P < 0.05), but that of the pHRE-Epo-treated rats didn't. Hypoxia-regulated system of Epo gene expression constructed by fusing Epo to the HRE/CMV promoter and delivered by plasmid intramuscular injection may provide a long-term and stable Epo expression and secretion in vivo to correct the anemia in adenine-induced uremic rats.


Sujet(s)
Animaux , Humains , Rats , Anémie/sang , Séquence nucléotidique , Azote uréique sanguin , Hypoxie cellulaire , Créatinine/sang , Érythropoïétine/biosynthèse , Régulation de l'expression des gènes , Gènes rapporteurs , Thérapie génétique , Cellules HeLa , Injections musculaires , Rein/anatomopathologie , Luciférases des lucioles/biosynthèse , Données de séquences moléculaires , Plasmides/génétique , Régions promotrices (génétique) , Rat Sprague-Dawley , Protéines recombinantes/biosynthèse , Éléments de réponse , Activation de la transcription , Urémie/sang
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