Antiapoptotic properties of recombinant human erythropoietin protects against tubular cyclosporine toxicity.
During the early post transplant period, the tubular epithelium is the main target of injuries including ischemia reperfusion and toxicity effects from calcineurin inhibitors. Taking into account the tissue protective effects of erythropoietin mediated through its antiapoptotic properties, we tested whether administration of recombinant human erythropoietin protects against acute cyclosporine nephrotoxicity. Four groups of five rats were intraperitoneally treated over 28 days with 100UI/Kg/48h Epoetin beta (15mg/kg/day CsA diluted in olive oil, 100UI/Kg/48h Epoetin beta+15mg/kg/day CsA, or olive oil. Histological changes due to tubular necrosis were evaluated with Masson'Trichrome staining. Apoptotic nuclei in kidneys were detected using the Terminal deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling (TUNEL) method. Phospho-Akt, Akt, cleaved caspase 3 and non cleaved caspase 3 expression were evaluated using immunblotting. We demonstrate that recombinant human erythropoietin (epoetin beta) improves renal function and protects against acute tubular injury. Our data suggest that this nephroprotective effect is mediated by Akt activation and inhibition of tubular apoptosis. Indeed, western blotting analysis of caspase 3 cleavage and Akt phosphorylation demonstrates that rhEPO activate Akt signaling and inhibits caspase 3 cleavage induced by CsA. TUNEL staining confirms that rhEPO inhibits CsA-induced tubular apoptosis. In conclusion, we describe here a new potential target of recombinant human erythropoietin and our results provide an interesting framework for further nephroprotective therapies based on recombinant human erythropoietin.
Pallet
Nicolas
N
INSERM U775 and Université Paris Descartes, Paris, France. nicolas.pallet@univ-paris5.fr
Bouvier
Nicolas
N
Legendre
Christophe
C
Beaune
Philippe
P
Thervet
Eric
E
Choukroun
Gabriel
G
Martinez
Frank
F
D000208:Acute Disease; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015415:Biomarkers / Q000097:blood; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D003404:Creatinine / Q000097:blood; D016572:Cyclosporine; D019610:Cytoprotection; D004195:Disease Models, Animal; D004921:Erythropoietin / Q000494:pharmacology*; D006801:Humans; D020287:In Situ Nick-End Labeling; D007673:Kidney Cortex Necrosis / Q000139:chemically induced / Q000473:pathology / Q000517:prevention & control*; D007684:Kidney Tubules / Q000187:drug effects* / Q000473:pathology; D008297:Male; D009336:Necrosis; D010766:Phosphorylation; D020011:Protective Agents / Q000494:pharmacology*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins; D015398:Signal Transduction / Q000187:drug effects
D015415:Biomarkers; D020011:Protective Agents; D011994:Recombinant Proteins; C103998:epoetin beta; D004921:Erythropoietin; D016572:Cyclosporine; D003404:Creatinine; D051057:Proto-Oncogene Proteins c-akt; C505410:Casp3 protein, rat; D053148:Caspase 3