Journal of Molecular Neuroscience

, Volume 61, Issue 3, pp 449–458 | Cite as

The Anti-Aging Effect of Erythropoietin via the ERK/Nrf2-ARE Pathway in Aging Rats

  • Haiqin Wu
  • Jiaxin Zhao
  • Mengyi Chen
  • Huqing Wang
  • Qingling Yao
  • Jiaxin Fan
  • Meng Zhang


Erythropoietin (EPO) has a neuroprotective effect and can resist aging, which most likely occur through EPO increasing the activity of antioxidant enzymes and scavenging free radicals. In this study, we verified the anti-aging function of EPO and discussed the mechanism occurring through the extracellular signal-regulated kinase (ERK)/NF-E2-related factor 2 (Nrf2)-ARE pathway. A rat model of aging was induced by the continuous subcutaneous injection of 5 % d-galactose for 6 weeks. At the beginning of the sixth week, physiological saline or EPO was administered twice per day through a lateral ventricle system for a total of 7 days. In one group, 2 μl PD98059 was administered 30 min before EPO. Learning and memory ability were analyzed with the Morris water maze system. HE staining was used to observe the morphological changes in the neurons in the hippocampus, and immunohistochemical staining as well as Western blots were carried out to detect the expression of ERK for each group of rats and the expression of phosphorylated-ERK (P-ERK), Nrf2, and superoxide dismutase (SOD). Real-Time PCR was carried out to detect the amount of Nrf2 mRNA and the KEAP1 mRNA expression. EPO can significantly improve learning and memory ability in aging rats and can provide protection against aging by improving the hippocampus morphology. Immunohistochemical staining and Western blots showed P-ERK, Nrf2, and Cu-Zn SOD decreases in aging rats compared to the normal group, while the expression for those proteins increased after EPO intervention. PD98059 inhibited the enhanced expression of P-ERK, Nrf2, and Cu-Zn SOD induced by EPO. Real-Time PCR results suggested that the trend of Nrf2mRNA expression was the same as that for the proteins, which confirmed that the enhancement occurred at the gene level. As such, EPO can significantly resist or delay aging and protect the brain by reducing oxidative stress. The most likely mechanism is that EPO can promote the ERK/Nrf2-ARE pathway in aging rats and that PD98059 can inhibit that process. These findings may facilitate further studies on the mechanism of aging and applications for the neuroprotective properties of EPO for clinical treatments.


EPO ERK P-ERK Nrf2 Cu-Zn SOD Anti-aging effects 



This study was supported by the National Natural Science Foundation of China (81170300).


  1. Buendia I, Michalska P, Navarro E, Gameiro I, Egea J, Leon R (2016) Nrf2-ARE pathway: an emerging target against oxidative stress and neuroinflammation in neurodegenerative diseases. Pharmacol Ther 157:84–104CrossRefPubMedGoogle Scholar
  2. Bundy JD, He J (2016) Hypertension and related cardiovascular disease burden in China. Ann Glob Health 82:227–233CrossRefPubMedGoogle Scholar
  3. Butler RN, Miller RA, Perry D, Carnes BA, Williams TF, Cassel C, Brody J, Bernard MA, Partridge L, Kirkwood T, Martin GM, Olshansky SJ (2008) New model of health promotion and disease prevention for the 21st century. BMJ 337:a399CrossRefPubMedGoogle Scholar
  4. Chetelat G, Ossenkoppele R, Villemagne VL, Perrotin A, Landeau B, Mezenge F, Jagust WJ, Dore V, Miller BL, Egret S, Seeley WW, van der Flier WM, La Joie R, Ames D, van Berckel BN, Scheltens P, Barkhof F, Rowe CC, Masters CL, de La Sayette V, Bouwman F, Rabinovici GD (2016) Atrophy, hypometabolism and clinical trajectories in patients with amyloid-negative Alzheimer’s disease. Brain 139:2528–2539CrossRefPubMedGoogle Scholar
  5. Costa LG, Garrick JM, Roque PJ, Pellacani C (2016) Mechanisms of neuroprotection by quercetin: counteracting oxidative stress and more. Oxidative Med Cell Longev 2016:2986796Google Scholar
  6. Digicaylioglu M, Lipton SA (2001) Erythropoietin-mediated neuroprotection involves cross-talk between Jak2 and NF-kappaB signalling cascades. Nature 412:641–647CrossRefPubMedGoogle Scholar
  7. Engel PA (2016) Is age-related failure of metabolic reprogramming a principal mediator in idiopathic Parkinson’s disease? Implications for treatment and inverse cancer risk. Med Hypotheses 93:154–160CrossRefPubMedGoogle Scholar
  8. Gassen NC, Chrousos GP, Binder EB, Zannas AS (2016) Life stress, glucocorticoid signaling, and the aging epigenome: implications for aging-related diseases. Neurosci Biobehav RevGoogle Scholar
  9. Genc K, Egrilmez MY, Genc S (2010) Erythropoietin induces nuclear translocation of Nrf2 and heme oxygenase-1 expression in SH-SY5Y cells. Cell Biochem Funct 28:197–201CrossRefPubMedGoogle Scholar
  10. Herrera-Arozamena C, Marti-Mari O, Estrada M, de la Fuente Revenga M, Rodriguez-Franco MI (2016) Recent advances in neurogenic small molecules as innovative treatments for neurodegenerative diseases. Molecules 21Google Scholar
  11. Jin W, Kong J, Lu T, Wang H, Ni H, Wu J, Dai Y, Jiang J, Liang W (2011) Erythropoietin prevents secondary brain injury induced by cortical lesion in mice: possible involvement of Nrf2 signaling pathway. Ann Clin Lab Sci 41:25–32PubMedGoogle Scholar
  12. Juul S (2012) Neuroprotective role of erythropoietin in neonates. J Matern Fetal Neonatal Med 25(Suppl 4):105–107PubMedGoogle Scholar
  13. Juul SE, Pet GC (2015) Erythropoietin and neonatal neuroprotection. Clin Perinatol 42:469–481CrossRefPubMedPubMedCentralGoogle Scholar
  14. Katsuoka F, Yamamoto M (2016) Small Maf proteins (MafF, MafG, MafK): history, structure and function. Gene 586:197–205CrossRefPubMedGoogle Scholar
  15. Katsuoka F, Motohashi H, Engel JD, Yamamoto M (2005) Nrf2 transcriptionally activates the mafG gene through an antioxidant response element. J Biol Chem 280:4483–4490CrossRefPubMedGoogle Scholar
  16. Khalil SK, Amer HA, El Behairy AM, Warda M (2016) Oxidative stress during erythropoietin hyporesponsiveness anemia at end stage renal disease: molecular and biochemical studies. J Adv Res 7:348–358CrossRefPubMedPubMedCentralGoogle Scholar
  17. Kilic E, Kilic U, Soliz J, Bassetti CL, Gassmann M, Hermann DM (2005) Brain-derived erythropoietin protects from focal cerebral ischemia by dual activation of ERK-1/-2 and Akt pathways. FASEB J 19:2026–2028PubMedGoogle Scholar
  18. Koltover VK (2016) Free radical timer of aging: from chemistry of free radicals to systems theory of reliability. Curr Aging SciGoogle Scholar
  19. Koskenkorva-Frank TS, Weiss G, Koppenol WH, Burckhardt S (2013) The complex interplay of iron metabolism, reactive oxygen species, and reactive nitrogen species: insights into the potential of various iron therapies to induce oxidative and nitrosative stress. Free Radic Biol Med 65:1174–1194CrossRefPubMedGoogle Scholar
  20. Kume T, Suenaga A, Izumi Y, Akaike A (2016) Protective effect of dimethyl fumarate on an oxidative stress model induced by sodium nitroprusside in mice. Biol Pharm Bull 39:1055–1059CrossRefPubMedGoogle Scholar
  21. Li YP, Yang GJ, Jin L, Yang HM, Chen J, Chai GS, Wang L (2015) Erythropoietin attenuates Alzheimer-like memory impairments and pathological changes induced by amyloid beta42 in mice. Brain Res 1618:159–167CrossRefPubMedGoogle Scholar
  22. Liochev SI (2015) Which is the most significant cause of aging? Antioxidants (Basel) 4:793–810CrossRefGoogle Scholar
  23. Lu MC, Ji JA, Jiang ZY, You QD (2016) The Keap1-Nrf2-ARE pathway as a potential preventive and therapeutic target: an update. Med Res Rev 36:924–963CrossRefPubMedGoogle Scholar
  24. Merelli A, Czornyj L, Lazarowski A (2013) Erythropoietin: a neuroprotective agent in cerebral hypoxia, neurodegeneration, and epilepsy. Curr Pharm Des 19:6791–6801CrossRefPubMedGoogle Scholar
  25. Merelli A, Czornyj L, Lazarowski A (2015) Erythropoietin as a new therapeutic opportunity in brain inflammation and neurodegenerative diseases. Int J Neurosci 125:793–797CrossRefPubMedGoogle Scholar
  26. Morishita E, Masuda S, Nagao M, Yasuda Y, Sasaki R (1997) Erythropoietin receptor is expressed in rat hippocampal and cerebral cortical neurons, and erythropoietin prevents in vitro glutamate-induced neuronal death. Neuroscience 76:105–116CrossRefPubMedGoogle Scholar
  27. Sanz A (2016) Mitochondrial reactive oxygen species: do they extend or shorten animal lifespan? Biochim Biophys Acta 1857:1116–1126CrossRefPubMedGoogle Scholar
  28. Sargin D, Friedrichs H, El-Kordi A, Ehrenreich H (2010) Erythropoietin as neuroprotective and neuroregenerative treatment strategy: comprehensive overview of 12 years of preclinical and clinical research. Best Pract Res Clin Anaesthesiol 24:573–594CrossRefPubMedGoogle Scholar
  29. Wu H, Wang H, Zhang W, Wei X, Zhao J, Yan P, Liu C (2015) rhEPO affects apoptosis in hippocampus of aging rats by upregulating SIRT1. Int J Clin Exp Pathol 8:6870–6880PubMedPubMedCentralGoogle Scholar
  30. Zhang DD (2006) Mechanistic studies of the Nrf2-Keap1 signaling pathway. Drug Metab Rev 38:769–789CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Haiqin Wu
    • 1
  • Jiaxin Zhao
    • 2
  • Mengyi Chen
    • 1
  • Huqing Wang
    • 1
  • Qingling Yao
    • 1
  • Jiaxin Fan
    • 1
  • Meng Zhang
    • 1
  1. 1.Department of Neurology, the Second Affiliated Hospital of Xi’anJiaotong UniversityXi’anPeople’s Republic of China
  2. 2.Department of NeurologyShaanxi Provincial People’s HospitalXi’anPeople’s Republic of China

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