Browse > Article
http://dx.doi.org/10.3345/kjp.2010.53.10.898

The neuroprotective effect of recombinant human erythropoietin via an antiapoptotic mechanism on hypoxic-ischemic brain injury in neonatal rats  

Kim, Moon-Sun (Department of Pediatrics, School of Medicine, Catholic University of Daegu)
Seo, Yoo-Kyung (Dongguk University College of Medicine)
Park, Hye-Jin (Department of Pediatrics, School of Medicine, Catholic University of Daegu)
Lee, Kye-Hyang (Department of Pediatrics, School of Medicine, Catholic University of Daegu)
Lee, Kyung-Hoon (Department of Pediatrics, School of Medicine, Catholic University of Daegu)
Choi, Eun-Jin (Department of Pediatrics, School of Medicine, Catholic University of Daegu)
Kim, Jin-Kyung (Department of Pediatrics, School of Medicine, Catholic University of Daegu)
Chung, Hai-Lee (Department of Pediatrics, School of Medicine, Catholic University of Daegu)
Kim, Woo-Taek (Department of Pediatrics, School of Medicine, Catholic University of Daegu)
Publication Information
Clinical and Experimental Pediatrics / v.53, no.10, 2010 , pp. 898-908 More about this Journal
Abstract
Purpose: The neuroprotective effects of erythropoietin (EPO) have been recently shown in many animal models of brain injury, including hypoxic-ischemic (HI) encephalopathy, trauma, and excitotoxicity; however, limited data are available for such effects during the neonatal periods. Therefore, we investigated whether recombinant human EPO (rHuEPO) can protect against perinatal HI brain injury via an antiapoptotic mechanism. Methods: The left carotid artery was ligated in 7-day-old Sprague-Dawley (SD) rat pups ($in$ $vivo$ model). The animals were divided into 6 groups: normoxia control (NC), normoxia sham-operated (NS), hypoxia only (H), hypoxia+vehicle (HV), hypoxia+rHuEPO before a hypoxic insult (HE-B), and hypoxia+rHuEPO after a hypoxic insult (HE-A). Embryonic cortical neuronal cell culture of SD rats at 18 days gestation ($in$ $vitro$ model) was performed. The cultured cells were divided into 5 groups: normoxia (N), hypoxia (H), and 1, 10, and 100 IU/mL rHuEPO-treated groups. Results: In the $in$ $vivo$ model, Bcl-2 expressions in the H and HV groups were lower than those in the NC and NS groups, whereas those in the HE-A and HE-B groups were greater than those of the H and HV groups. The expressions of Bax and caspase-3 and the ratio of Bax/Bcl-2 were in contrast to those of Bcl-2. In the $in$ $vitro$ model, the patterns of Bcl-2, Bax, and caspase-3 expression and Bax/Bcl-2 ratio were similar to the results obtained in the in vivo model. Conclusion: rHuEPO exerts neuroprotective effect against perinatal HI brain injury via an antiapoptotic mechanism.
Keywords
Erythropoietin; Apoptosis; Hypoxic-ischemic brain injury; Bcl-2; Bax; Caspase-3;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Markianos M, Kosmidis ML, Sfagos C. Reductions in plasma prolactin during acute erythropoietin administration. Neuro Endocrinol Lett 2006;27:355-8.
2 Kawakami M, Sekiguchi M, Sato K, Kozaki S, Takahashi M. Erythropoietin receptor-mediated inhibition of exocytotic glutamate release confers neuroprotection during chemical ischemia. J Biol Chem 2001;276:39469-75.   DOI   ScienceOn
3 Pulera MR, Adams LM, Liu H, Santos DG, Nishimura RN, Yang F, et al. Apoptosis in a neonatal rat model of cerebral hypoxia-ischemia. Stroke 1998;29:2622-30.   DOI   ScienceOn
4 Carlini RG, Alonzo EJ, Dominguez J, Blanca I, Weisinger JR, Rothstein M, et al. Effect of recombinant human erythropoietin on endothelial cell apoptosis. Kidney Int 1999;55:546-53.   DOI   ScienceOn
5 Juul SE, Yachnis AT, Rojiani AM, Christensen RD. Immunohistochemical localization of erythropoietin and its receptor in the developing human brain. Pediatr Dev Pathol 1999;2:148-58.   DOI   ScienceOn
6 Sinor AD and Greenberg DA. Erythropoietin protects cultured cortical neurons, but not astroglia, from hypoxia and AMPA toxicity. Neurosci Lett 2000;290:213-5.   DOI   ScienceOn
7 Brines M and Cerami A. Emerging biological roles for erythropoietin in the nervous system. Nat Rev Neurosci 2005;6:484-94.   DOI   ScienceOn
8 Grasso G, Sfacteria A, Cerami A, Brines M. Erythropoietin as a tissueprotective cytokine in brain injury: what do we know and where do we go? Neuroscientist 2004;10:93-8.   DOI   ScienceOn
9 Yu X, Shacka JJ, Eels JB, Suarez-Quinan C, Przygodski RM, Beleslin- Cokic B, et al. Erythropoietin receptor signalling is required for normal brain development. Development 2002;129:505-16.
10 Wang L, Zhang Z, Wang Y, Zhang R, Chopp M. Treatment of stroke with erythropoietin enhances neurogenesis and angiogenesis and improves neurological function in rats. Stroke 2004;35:1732-7.   DOI   ScienceOn
11 Ribatti D, Vacca A, Roccaro AM, Crivellato E, Presta M. Erythropoietin as an angiogenic factor. Eur J Clin Invest 2003;33:891-6.   DOI   ScienceOn
12 Sugawa M, Sakurai Y, Ishikawa-Ieda Y, Suzuki H, Asou H. Effects of erythropoietin on glial cell development; oligodendrocyte maturation and astrocyte proliferation. Neurosci Res 2002;44:391-403.   DOI   ScienceOn
13 Villa P, Bigini P, Mennini T, Agnello D, Laragione T, Cagnotto A, et al. Erythropoietin selectively attenuates cytokine production and inflammation in cerebral ischemia by targeting neuronal apoptosis. J Exp Med 2003;198:971-5.   DOI   ScienceOn
14 Anderson P, Doyle LW; Victorian Infant Collaborative Study Group. Neurobehavioral outcomes of school-age children born extremely low birth weight or very preterm in the 1990s. JAMA 2003;289:3264-72.   DOI   ScienceOn
15 Koshimura K, Murakami Y, Sohmiya M, Tanaka J, Kato Y. Effects of erythropoietin on neuronal activity. J Neurochem 1999;72:2565-72.
16 Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.   DOI   ScienceOn
17 Vannucci RC and Perlman JM. Interventions for perinatal hypoxicischemic encephalopathy. Pediatrics 1997;100:1004-14.   DOI   ScienceOn
18 Jelkmann W. Erythropoietin: structure, control of production, and function. Physiol Rev 1992;72:449-89.   DOI
19 Siren AL, Fratelli M, Brines M, Goemans C, Casagrande S, Lewczuk P, et al. Erythropoietin prevents neuronal apoptosis after cerebral ischemia and metabolic stress. Proc Natl Acad Sci USA 2001;98:4044-9.   DOI   ScienceOn
20 Bernaudin M, Marti HH, Roussel S, Divoux D, Nouvelot A, MacKenzie ET, et al. A potential role for erythropoietin in focal permanent cerebral ischemia in mice. J Cereb Blood Flow Metab 1999;19:643-51.   DOI
21 Spandou E, Soubasi V, Papoutsopoulou S, Karkavelas G, Simeonidou C, Kaiki-Astara A, et al. Erythropoietin prevents hypoxia/ischemia-induced DNA fragmentation in an experimental model of perinatal asphyxia. Neurosci Lett 2004;366:24-8.   DOI   ScienceOn
22 Ruscher K, Freyer D, Karsch M, Isaev N, Megow D, Sawitzki B, et al. Erythropoietin is a paracrine mediator of ischemic tolerance in the brain, evidence from an in vitro model. J Neurosci 2002;22:10291-301.
23 Volpe JJ. Perinatal brain injury: from pathogenesis to neuroprotection. Ment Retard Dev Disabil Res Rev 2001;7:56-64   DOI   ScienceOn
24 Dzietko M, Felderhoff-Mueser U, Sifringer M, Krutz B, Bittigau P, Thor F, et al. Erythropoietin protects the developing brain against N-methyl-daspartate receptor antagonist neurotoxicity. Neurobiol Dis 2004;15:177- 87.   DOI   ScienceOn
25 Morishita E, Masuda S, Nagao M, Tasuda Y, Sasaki R. Erythropoietin receptor is expressed in rat hippocampal and cerebral cortical neurons, and erythropoietin prevents in vitro glutamateinduced neuronal death. Neuroscience 1997;76:105-16.
26 Marti HH, Wenger RH, Rivas LA, Straumann U, Digicaylioglu M, Hehn V, et al. Erythropoietin gene expression in human monkey and murine brain. Eur J Neurosci 1996;8:666-76.   DOI   ScienceOn
27 Chong ZZ, Kang JQ, Maiese K. Hematopoietic factor erythropoietin fosters neuroprotection through novel signal transduction cascades. J Cereb Blood Flow Metab 2002;22:503-14.   DOI
28 Vairano M, Russo CD, Pozzoli G, Battaglia A, Scambia G, Tringali G, et al. Erythropoietin exerts antiapoptotic effects on rat microglial cells in vitro . Eur J Neurosci 2002;16:584-92.   DOI   ScienceOn
29 Wen TC, Sadamoto Y, Tanaka J, Zhu PX, Nakata K, Ma YJ, et al. Erythropoietin protects neurons against chemical hypoxia and cerebral ischemic injury by upregulating Bcl-xL expression. J Neurosci Res 2002;67:795-803.   DOI   ScienceOn
30 Yamasaki M, Mishima HK, Yamashita H, Kashiwagi K, Murata K, Minamoto A, et al. Neuroprotective effects of erythropoietin on glutamate and nitric oxide toxicity in primary cultured retinal ganglion cells. Brain Res 2005;1050:15-26.   DOI   ScienceOn
31 Feng Y, Fratkin JD, LeBlanc MH. Treatment with tamoxifen reduces hypoxic-ischemic brain injury in neonatal rats. Eur J Pharmacol 2004;484:65-74.   DOI   ScienceOn
32 Emir M, Ozisik K, Cagli K, Misirlioglu M, Ozisik P, Iscan Z, et al. Effect of erythropoietin on bcl-2 gene expression in rat cardiac myocytes after traumatic brain injury. Transplant Proc 2004;36:2935-8.   DOI   ScienceOn
33 Sakanaka M, Wen TC, Matsuda S, Masuda S, Morishita E, Nagao M, et al. In vivo evidence that erythropoietin protects neurons from ischemic damage. Proc Natl Acad Sci USA 1998;95:4635-40.   DOI   ScienceOn
34 Brewer GJ. Isolation and culture of adult rat hippocampal neurons. J Neurosci Methods 1997;71:143-55.   DOI   ScienceOn
35 Dame C, Juul S, Christensen RD. The biology of erythropoietin in the central nervous system and its neurotrophic and neuroprotective potential. Biol Neonate 2001;79:228-35.   DOI   ScienceOn
36 Digicaylioglu M, Bichet S, Marti HH, Wenger RH, Rivas LA, Bauer C, et al. Localization of specific erythropoietin binding sites in defined areas of the mouse brain. Proc Natl Acad Sci USA 1995;92:3717-20.   DOI   ScienceOn
37 Ghezzi P and Brines M. Erythropoietin as an antiapoptotic, tissueprotective cytokine. Cell Death Differ 2004;11:S37-44.   DOI   ScienceOn
38 Chong ZZ, Lin SH, Kang JQ, Maiese K. Erythropoietin prevents early and late neuronal demise through modulation of Akt1 and induction of caspase 1, 3, and 8. J Neurosci Res 2003;71:659-69.   DOI   ScienceOn
39 Liu R, Suzuki A, Guo Z, Mizuno Y, Urabe T. Intrinsic and extrinsic erythropoietin enhances neuroprotection against ischemia and reperfusion injury in vitro . J Neurochem 2006;96:1101-10.   DOI   ScienceOn
40 Rice JE 3rd, Vannucci RC, Brierley JB. The influence of immaturity on hypoxic-ischemic brain damage in the rat. Ann Neurol 1981;9:131-41.   DOI   ScienceOn