Protective Effect of Erythropoietin against Radiocontrast-Induced Renal Tubular Epithelial Cell Injury

Abstract

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.