Antiapoptotic properties of erythropoiesis-stimulating proteins in models of cisplatin-induced acute kidney injury.
Erythropoietin (Epo) induces erythrocytosis by suppressing erythroid progenitor cell apoptosis through the Janus-activated kinase-signal transducers and activators of transcription (JAK-STAT) pathway. Since apoptosis contributes to cisplatin (CP)-induced nephrotoxicity and Epo receptors (EpoR) are expressed in the kidney, we examined the role of antiapoptosis in recombinant human erythropoietin (rHuEpo)-mediated renal protection. In human renal proximal tubular epithelial (RPTE) cells in culture, rHuEpo, but not inactive rHuEpo (I-rHuEpo), the receptor-binding sites of which are mutated, caused a significant reduction in CP-induced apoptosis at > or = 100 U/ml. rHuEpo, but not I-rHuEpo, increased STAT5 and Akt/PKB phosphorylation, demonstrating functional EpoR expression on RPTE cells. Furthermore, the JAK2 inhibitor tyrphostin AG-490 attenuated rHuEpo protection, suggesting a role of the JAK-STAT pathway in rHuEpo-mediated antiapoptosis. In rats, intravenous administration of 5,000 U/kg rHuEpo, but not an equivalent peptide mass of I-rHuEpo, before a single 5.5 mg/kg iv injection of CP, significantly increased hematocrit (Hct) and reduced the CP-induced increase in serum creatinine. Serum creatinine on day 4 was 3.4 +/- 0.3, 1.9 +/- 0.3, and 3.5 +/- 0.4 mg/dl in the CP, CP + rHuEpo, and CP + I-rHuEpo groups, respectively. Similarly, darbepoietin-alpha (DA), a hyperglycosylated analog of rHuEpo with prolonged in vivo activity when injected at 25 microg/kg iv before CP, significantly increased Hct and reduced serum creatinine. Renal clearance studies based on glomerular filtration rate and renal blood flow confirmed the significant renal protection by DA against CP. Tubular apoptosis and necrosis were significantly reduced in the kidneys of the CP + DA vs. the CP + saline group. Moreover, the equalization of Hct by venesection did not abrogate the DA-mediated renal protection. Administration of DA 48 h after CP injection also conferred significant renal protection. Thus our experiments confirm a role for erythropoiesis-stimulating proteins, including the new analog DA, in limiting CP-induced nephrotoxicity and suggest that antiapoptosis via the Epo-EpoR interaction is an important mechanism for renal protection.
Salahudeen
Abdulla K
AK
Department of Medicine, University of Mississippi Medical Center, Jackson, MI, , USA. Aksalahu@mdanderson.org
Haider
Naeem
N
Jenkins
John
J
Joshi
Manish
M
Patel
Harnish
H
Huang
Hong
H
Yang
Ming
M
Zhe
He
H
D058186:Acute Kidney Injury / Q000139:chemically induced* / Q000188:drug therapy* / Q000473:pathology; D000818:Animals; D000970:Antineoplastic Agents / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D002478:Cells, Cultured; D002945:Cisplatin / Q000633:toxicity*; D000068256:Darbepoetin alfa; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004921:Erythropoietin / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology*; D006397:Hematinics / Q000378:metabolism / Q000494:pharmacology; D006400:Hematocrit; D006801:Humans; D053614:Janus Kinase 2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D007668:Kidney / Q000166:cytology; D008297:Male; D009336:Necrosis / Q000378:metabolism / Q000473:pathology; D010766:Phosphorylation / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D017467:Receptors, Erythropoietin / Q000378:metabolism; D011994:Recombinant Proteins; D050799:STAT5 Transcription Factor / Q000378:metabolism; D020032:Tyrphostins / Q000494:pharmacology
D000970:Antineoplastic Agents; D004791:Enzyme Inhibitors; D006397:Hematinics; D017467:Receptors, Erythropoietin; D011994:Recombinant Proteins; D050799:STAT5 Transcription Factor; D020032:Tyrphostins; C095512:alpha-cyano-(3,4-dihydroxy)-N-benzylcinnamide; D004921:Erythropoietin; D000068256:Darbepoetin alfa; D053614:Janus Kinase 2; C494920:Akt1 protein, rat; D051057:Proto-Oncogene Proteins c-akt; D002945:Cisplatin