Background/Aims: Recombinant human erythropoietin (rhEpo) has been shown to reduce tissue injury following ischemia-reperfusion. We examined whether rhEpo protects in vitro renal tubular epithelial cells against radiocontrast media-induced injury. Methods: LLC-PK1 renal tubular epithelial cells were exposed to non-ionic radiocontrast agent iohexol (low-osmolar) or iodixanol (iso-osmolar), with or without rhEpo (200 U/ml). Following a 6-hour exposure, cells were incubated for 24 h in radiocontrast-free culture medium. Cell viability was then assessed by the MTT assay. We also assessed cell apoptosis by the TUNEL assay, and activities of caspase-3, caspase-8, and caspase-9 were determined by a luminescence assay. Results: rhEpo improved viability of iohexol-treated LLC-PK1 cells by 27 ± 6% (88.1 ± 1.5 vs. 70.8 ± 3.3%, p = 0.008). Similarly, rhEpo improved the viability of iodixanol-treated LLC-PK1 cells by 26 ± 4% (82.5 ± 2.1vs. 65.7 ± 1.7%, p = 0.028). rhEpo also decreased apoptosis rates of iohexol-treated LLC-PK1 cells (6.4 ± 0.9/1,000 cells vs. 14.8 ± 2.4/1,000 cells, p = 0.028), and iodixanol-treated LLC-PK1 cells (8.0 ± 1.2/1,000 cells vs. 13.5 ± 1.9/1,000 cells, p = 0.028). In iohexol-treated LLC-PK1 cells, rhEpo attenuated activation of caspase-3 (p = 0.003), caspase-8 (p = 0.033) and caspase-9 (p = 0.055). Conclusion: rhEpo attenuates in vitro renal tubular epithelial cell injury induced by low- and iso-osmolar radiocontrast media, possibly by reduction of caspases activation and apoptosis rates.

Levy EM, Viscoli CM, Horwitz RI: The effect of acute renal failure on mortality: a cohort analysis. JAMA 1996;275:1489–1494.
McCullough PA, Wolyn R, Rocher LL, Levin RN, O’Neill WW: Acute renal failure after coronary intervention: incidence, risk factors, and relationship to mortality. Am J Med 1997;103:368–375.
Rihal CS, Textor SC, Grill DE, et al: Incidence and prognostic importance of acute renal failure after percutaneous coronary intervention. Circulation 2002;105:2259–2264.
Mehran R, Aymong ED, Nikolsky E, et al: A simple risk score for prediction of contrast-induced nephropathy after percutaneous coronary intervention: development and initial validation. J Am Coll Cardiol 2004;44:1393–1399.
Mehran R, Nikolsky E: Contrast-induced nephropathy: definition, epidemiology, and patients at risk. Kidney Int Suppl 2006;100:S11–S15.
Zager RA, Johnson AC, Hanson SY: Radiographic contrast media-induced tubular injury: evaluation of oxidant stress and plasma membrane integrity. Kidney Int 2003;64:128–139.
Itoh Y, Yano T, Sendo T, Oishi R: Clinical and experimental evidence for prevention of acute renal failure induced by radiographic contrast media. J Pharmacol Sci 2005;97:473–488.
Barrett BJ, Parfrey PS: Clinical practice. Preventing nephropathy induced by contrast medium. N Engl J Med 2006;354:379–386.
Solomon R, Werner C, Mann D, D’Elia J, Silva P: Effects of saline, mannitol, and furosemide to prevent acute decreases in renal function induced by radiocontrast agents. N Engl J Med 1994;331:1416–1420.
Merten GJ, Burgess WP, Gray LV, et al: Prevention of contrast-induced nephropathy with sodium bicarbonate: a randomized controlled trial. JAMA 2004;291:2328–2334.
Alonso A, Lau J, Jaber BL, Weintraub A, Sarnak MJ: Prevention of radiocontrast nephropathy with N-acetylcysteine in patients with chronic kidney disease: a meta-analysis of randomized, controlled trials. Am J Kidney Dis 2004;43:1–9.
Aspelin P, Aubry P, Fransson SG, Strasser R, Willenbrock R, Berg KJ: Nephrotoxic effects in high-risk patients undergoing angiography. N Engl J Med 2003;348:491–499.
Porter AG, Janicke RU: Emerging roles of caspase-3 in apoptosis. Cell Death Differ 1999;6:99–104.
Yang B, El Nahas AM, Thomas GL, et al: Caspase-3 and apoptosis in experimental chronic renal scarring. Kidney Int 2001;60:1765–1776.
Perianayagam MC, Balakrishnan VS, Pereira BJ, Jaber BL: C5a delays apoptosis of human neutrophils via an extracellular signal-regulated kinase and Bad-mediated signalling pathway. Eur J Clin Invest 2004;34:50–56.
Yano T, Itoh Y, Sendo T, Kubota T, Oishi R: Cyclic AMP reverses radiocontrast media-induced apoptosis in LLC-PK1 cells by activating A kinase/PI3 kinase. Kidney Int 2003;64:2052–2063.
Yano T, Itoh Y, Kubota T, Sendo T, Oishi R: A prostacyclin analog beraprost sodium attenuates radiocontrast media-induced LLC-PK1 cells injury. Kidney Int 2004;65:1654–1663.
Yano T, Itoh Y, Kubota T, et al: A prostacyclin analog prevents radiocontrast nephropathy via phosphorylation of cyclic AMP response element binding protein. Am J Pathol 2005;166:1333–1342.
Zager RA, Burkhart KM: Differential effects of glutathione and cysteine on Fe2+, Fe3+, H2O2 and myoglobin-induced proximal tubular cell attack. Kidney Int 1998;53:1661–1672.
Lorusso V, Taroni P, Alvino S, Spinazzi A: Pharmacokinetics and safety of iomeprol in healthy volunteers and in patients with renal impairment or end-stage renal disease requiring hemodialysis. Invest Radiol 2001;36:309–316.
Donnelly PK, Burwell N, McBurney A, Ward JW, Walls J, Watkin EM: Hemodialysis and iopamidol clearance after subclavian venography. Invest Radiol 1993;28:629–632.
Eschbach JW, Egrie JC, Downing MR, Browne JK, Adamson JW: Correction of the anemia of end-stage renal disease with recombinant human erythropoietin. Results of a combined phase I and II clinical trial. N Engl J Med 1987;316:73–78.
Eschbach JW, Kelly MR, Haley NR, Abels RI, Adamson JW: Treatment of the anemia of progressive renal failure with recombinant human erythropoietin. N Engl J Med 1989;321:158–163.
Adamson JW, Eschbach JW: Erythropoietin for end-stage renal disease. N Engl J Med 1998;339:625–627.
Peterson TE, Katusic ZS: EPO tecting the endothelium. Br J Pharmacol 2007;150:823–825.
Boogaerts M: Pleiotropic effects of erythropoietin in neuronal and vascular systems. Curr Med Res Opin 2006;22(suppl 4):15–22.
Koury MJ, Bondurant MC: Erythropoietin retards DNA breakdown and prevents programmed death in erythroid progenitor cells. Science 1990;248:378–381.
Bittorf T, Seiler J, Ludtke B, Buchse T, Jaster R, Brock J: Activation of STAT5 during EPO-directed suppression of apoptosis. Cell Signal 2000;12:23–30.
Bittorf T, Jaster R, Brock J: Rapid activation of the MAP kinase pathway in hematopoietic cells by erythropoietin, granulocyte-macrophage colony-stimulating factor and interleukin-3. Cell Signal 1994;6:305–311.
Haseyama Y, Sawada K, Oda A, et al: Phosphatidylinositol 3-kinase is involved in the protection of primary cultured human erythroid precursor cells from apoptosis. Blood 1999;94:1568–1577.
Renzi MJ, Farrell FX, Bittner A, et al: Erythropoietin induces changes in gene expression in PC-12 cells. Brain Res Mol Brain Res 2002;104:86–95.
Vairano M, Dello Russo C, Pozzoli G, et al: Erythropoietin exerts anti-apoptotic effects on rat microglial cells in vitro. Eur J Neurosci 2002;16:584–592.
Wen TC, Sadamoto Y, Tanaka J, et al: Erythropoietin protects neurons against chemical hypoxia and cerebral ischemic injury by up-regulating Bcl-xL expression. J Neurosci Res 2002;67:795–803.
Sharples EJ, Patel N, Brown P, et al: Erythropoietin protects the kidney against the injury and dysfunction caused by ischemia- reperfusion. J Am Soc Nephrol 2004;15:2115–2124.
Buemi M, Vaccaro M, Sturiale A, et al: Recombinant human erythropoietin influences revascularization and healing in a rat model of random ischaemic flaps. Acta Derm Venereol 2002;82:411–417.
Bagnis C, Beaufils H, Jacquiaud C, et al: Erythropoietin enhances recovery after cisplatin-induced acute renal failure in the rat. Nephrol Dial Transplant 2001;16:932–938.
Vesey DA, Cheung C, Pat B, Endre Z, Gobe G, Johnson DW: Erythropoietin protects against ischaemic acute renal injury. Nephrol Dial Transplant 2004;19:348–355.
Johnson DW, Pat B, Vesey DA, Guan Z, Endre Z, Gobe GC: Delayed administration of darbepoetin or erythropoietin protects against ischemic acute renal injury and failure. Kidney Int 2006;69:1806–1813.
Brines M, Cerami A: Discovering erythropoietin’s extrahematopoietic functions: biology and clinical promise. Kidney Int 2006;70:246–250.
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.