Carbamylated Erythropoietin Ameliorate the Transforming Growth Factor Beta-1-Induced Epithelial-to-Mesenchymal Transition
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Recombinant human erythropoietin (EPO), a glycohormone, is one of the leading biopharmaceutical products, while carbamylated erythropoietin (CEPO), an EPO derivative, is attracting widespread interest due to its neuroprotective effects without erythropoiesis in several cells and animal models. However, the protective effects of CEPO on renal fibrosis still remain unknown. In this study, we evaluated the inhibitory effects of CEPO against transforming growth factor (TGF)-β1-induced epithelial-to-mesenchymal transition (EMT) in Madin-Darby Canine Kidney (MDCK) cells. MDCK cells were treated with TGF-β1 (5 ng/mL) for 48 h to induce EMT, and the cells were then co-treated with TGF-β1 and CEPO or recombinant human EPO for another 48 h. Increased expressions of α-smooth muscle actin (α-SMA) and decreased expressions of E-cadherin were observed after TGF-β1 treatment, and these changes were markedly attenuated by CEPO co-treatment. TGF-β1 increased phosphorylated Smad-2 expression in MDCK cells, which was decreased by CEPO cotreatment. The results indicate that CEPO ameliorates the TGF-β1-induced EMT mediated by p-Smad2 pathway. It suggests that CEPO has therapeutic potential in chronic kidney disease.
Abstract
Recombinant human erythropoietin (EPO), a glycohormone, is one of the leading biopharmaceutical products, while carbamylated erythropoietin (CEPO), an EPO derivative, is attracting widespread interest due to its neuroprotective effects without erythropoiesis in several cells and animal models. However, the protective effects of CEPO on renal fibrosis still remain unknown. In this study, we evaluated the inhibitory effects of CEPO against transforming growth factor (TGF)-β1-induced epithelial-to-mesenchymal transition (EMT) in Madin-Darby Canine Kidney (MDCK) cells. MDCK cells were treated with TGF-β1 (5 ng/mL) for 48 h to induce EMT, and the cells were then co-treated with TGF-β1 and CEPO or recombinant human EPO for another 48 h. Increased expressions of α-smooth muscle actin (α-SMA) and decreased expressions of E-cadherin were observed after TGF-β1 treatment, and these changes were markedly attenuated by CEPO co-treatment. TGF-β1 increased phosphorylated Smad-2 expression in MDCK cells, which was decreased by CEPO cotreatment. The results indicate that CEPO ameliorates the TGF-β1-induced EMT mediated by p-Smad2 pathway. It suggests that CEPO has therapeutic potential in chronic kidney disease.
참고문헌 (29)
- Bunn HF, "Oxygen sensing and molecular adaptation to hypoxia", Physiol Rev, 76, pp.839~885 (1996)
- Lacombe C, "Biology of erythropoietin", Haematologica, 83, pp.724~732 (1998)
- Diskin CJ, "Beyond anemia : the clinical impact of the physiologic effects of erythropoietin", Semin Dial, 21, pp.447~454 (2008)
- Brines M, "Erythropoietin mediates tissue protection through an erythropoietin and common beta-subunit heteroreceptor", Proc Natl Acad Sci USA, 101, pp.14907~14912 (2004)
- Moon C, "Erythropoietin, modified to not stimulate red blood cell production, retains its cardioprotective properties", J Pharmacol Exp Ther, 316, pp.999~1005 (2006)
- Coleman TR, "Cytoprotective doses of erythropoietin or carbamylated erythropoietin have markedly different procoagulant and vasoactive activities", Proc Natl Acad Sci USA, 103, pp.5965~5970 (2006)
- Leist M, "Derivatives of erythropoietin that are tissue protective but not erythropoietic", Science, 305, pp.239~242 (2004)
- Adembri C, "Carbamylated erythropoietin is neuroprotective in an experimental model of traumatic brain injury", Crit Care Med, 36, pp.975~978 (2008)
- Fiordaliso F, "A nonerythropoietic derivative of erythropoietin protects the myocardium from ischemia-reperfusion injury", Proc Natl Acad Sci USA, 102, pp.2046~2051 (2005)
- Imamura R, "Carbamylated erythropoietin improves angiogenesis and protects the kidneys from ischemia-reperfusion injury", Cell Transplant, 17(1-2), pp.135~141 (2008)
- Brines M, "Discovering erythropoietin’s extra-hematopoietic functions : biology and clinical promise", Kidney Int, 70, pp.246~250 (2006)
- Iwano M, "Evidence that fibroblasts derive from epithelium during tissue fibrosis", J Clin Inves, 110, pp.341~350 (2002)
- Strutz F, "Identification and characterization of a fibroblast marker : FSP1", J Cell Biol, 130, pp.393~405 (1995)
- Yang J, "Dissection of key events in tubular epithelial to myofibroblast transition and its implications in renal interstitial fibrosis", Am J Pathol, 159, pp.1465~1475 (2001)
- Kalluri R, "Epithelial-mesenchymal transition and its implications for fibrosis", J Clin Invest, 112, pp.1776~1784 (2003)
- Fan JM, "Transforming growth factor-β regulates tubular epithelial-myofibroblast transdifferentiation in vitro", Kidney Int, 56, pp.1455~1467 (1999)
- Gouva C, "Anemia early in renal failure patients slows the decline of renal function : a randomized controlled trial", Kidney Int, 66, pp.753~760 (2004)
- Kirkeby A, "High-dose erythropoietin alters platelet reactivity and bleeding time in rodents in contrast to the neuroprotective variant carbamyl-erythropoietin(CEPO)", Thromb Haemost, 99, pp.720~728 (2008)
- Imamura R, "Carbamylated erythropoietin protects the kidneys from ischemiareperfusion injury without stimulating erythropoiesis", Biochem Biophys Res Commun, 353, pp.786~792 (2007)
- Kitamura H, "Nonerythropoietic derivative of erythropoietin protects against tubulointerstitial injury in a unilateral ureteral obstruction model", Nephrol Dial Transplant, 23, pp.1521~1528 (2008)
- Kalluri R, "The basics of epithelial-mesenchymal transition", J Clin Invest, 119, pp.1420~1428 (2009)
- Ueno T, "Mesenchymal stem cells ameliorate experimental peritoneal fibrosis by suppressing inflammation and inhibiting TGF-β1 signaling", Kidney Int, 84, pp.297~307 (2013)
- Vesey DA, "Erythropoietin protects against ischaemic acute renal injury", Nephrol Dial Transplant, 19, pp.348~355 (2004)
- Park SH, "Erythropoietin decreases renal fibrosis in mice with ureteral obstruction : role of inhibiting TGF-beta-induced epithelial-tomesenchymal transition", J Am Soc Nephrol, 18, pp.1497~1507 (2007)
- Ding J, "Carbamylated erythropoietin ameliorates hypoxia-induced cognitive and behavioral defects with the generation of choline acetyltransferase-positive neurons", J Neurosci Res, 91, pp.73~82 (2013)
- Kirkeby A, "High-dose erythropoietin alters platelet reactivity and bleeding time in rodents in contrast to the neuroprotective variant carbamyl-erythropoietin(CEPO)", Thromb Haemost, 99, pp.720~728 (2008)
- Vaziri ND, "Erythropoietin enhances recovery from cisplatin-induced acute renal failure", Am J Physiol, 266, pp.F360~F366 (1994)
- Chatterjee PK, "Pleiotropic renal actions of erythropoietin", Lancet, 365, pp.1890~1892 (2005)
- Bahlmann FH, "Low-dose therapy with the long-acting erythropoietin analogue darbepoetin alpha persistently activates endothelial Akt and attenuates progressive organ failure", Circulation, 110, pp.1006~1012 (2004)