Long-term and stable correction of uremic anemia by intramuscular injection of plasmids containing hypoxia-regulated system of erythropoietin expression.
- Author:
Jifeng SUN
1
;
Yarong WANG
;
Jie YANG
;
Dewei DU
;
Zhanting LI
;
Junxia WEI
;
Angang YANG
Author Information
1. Department of Nephrology, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China. jifeng-sun@medmail.com.cn, yangjie72029@yahoo.com.cn
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
anemia;
erythropoietin;
gene therapy;
hypoxia response element;
uremia
- MeSH:
Anemia/blood/*therapy;
Animals;
Base Sequence;
Blood Urea Nitrogen;
Cell Hypoxia;
Creatinine/blood;
Erythropoietin/biosynthesis/*genetics/secretion;
Gene Expression Regulation;
Genes, Reporter;
Genetic Therapy;
HeLa Cells;
Humans;
Injections, Intramuscular;
Kidney/pathology;
Luciferases, Firefly/biosynthesis/genetics;
Molecular Sequence Data;
Plasmids/*genetics;
Promoter Regions, Genetic;
Rats;
Rats, Sprague-Dawley;
Recombinant Proteins/biosynthesis/genetics/secretion;
Response Elements;
Transcriptional Activation;
Uremia/blood/*therapy
- From:Experimental & Molecular Medicine
2012;44(11):674-683
- CountryRepublic of Korea
- Language:English
-
Abstract:
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.
