Abstract
The aim of this study was to explore the neuroprotective effect and the underlying mechanism of erythropoietin (EPO) on the cortical neuronal cells insulted with oxygen and glucose deprivation (OGD). Different concentrations of EPO were used to determine the anti-apoptosis effect of EPO. In addition, PI3K inhibitor LY294002 and ERK1/2 inhibitor U0126 were added to explore the underlying mechanism of EPO. Cell apoptosis rate was measured by flow cytometry. The protein expression of Bax, Bcl-2, cleaved caspase-3, AKT, p-AKT, Erk1/2 and p-Erk1/2 wasmeasured by Western blot. Our results showed that EPO alleviates OGD-induced cell apoptosis in a dose-dependent manner; the neuroprotective effect of EPO was further confirmed by the fact that EPO treatment reversed the protein expression of cleaved caspase-3, as well as the Bcl-2/Bax ratio as compared with the OGD treatment. In the mechanism part, our results demonstrated that OGD and EPO nearly had no influence on the protein expression of AKT and Erk1/2 but altered the phosphorylation of them. Specifically, OGD decreased the expression of p-AKT and increased the expression of p-Erk1/2; while, EPO treatment reversed the expression of p-AKT and p-Erk1/2 as compared with OGD treatment. Interestingly, LY294002 decreased the expression of p-AKT and attenuated the neuroprotective effect of EPO; while, U0126 decreased the expression of p-Erk1/2 and enhanced the neuroprotective effect of EPO. Our study demonstrated that EPO protects neurons against apoptosis induced by OGD, which is closely related with activation of PI3K/AKT and inactivation of Erk1/2 signaling pathway.
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References
Biggar P, Kim GH (2017) Treatment of renal anemia: erythropoiesis stimulating agents and beyond. Kidney Res Clin Pract 36:209–223. https://doi.org/10.23876/j.krcp.2017.36.3.209
Castillo C et al (2017) Neuroprotective effect of a new variant of Epo nonhematopoietic against oxidative stress. Redox Biol 14:285–294. https://doi.org/10.1016/j.redox.2017.09.010
Chen T et al (2017) Sirt1–Sirt3 axis regulates human blood–brain barrier permeability in response to ischemia. Redox Biol 14:229–236. https://doi.org/10.1016/j.redox.2017.09.016
Ding J et al (2017) Neuroprotection and CD131/GDNF/AKT pathway of carbamylated erythropoietin in hypoxic neurons. Mol Neurobiol 54:5051–5060. https://doi.org/10.1007/s12035-016-0022-0
Garg BD, Sharma D, Bansal A (2017) Systematic review seeking erythropoietin role for neuroprotection in neonates with hypoxic ischemic encephalopathy: presently where do we stand. J Matern Fetal Neonatal Med. https://doi.org/10.1080/14767058.2017.1366982
Gong G et al (2014) Tetramethylpyrazine suppresses transient oxygen-glucose deprivation-induced connexin32 expression and cell apoptosis via the ERK1/2 and p38 MAPK pathway in cultured hippocampal neurons. PLoS One 9:e105944. https://doi.org/10.1371/journal.pone.0105944
Grasso G, Alafaci C, Buemi M (2015) Erythropoietin in traumatic brain injury: an answer will come soon. World Neurosurg 84:1491–1492. https://doi.org/10.1016/j.wneu.2015.05.056
Guangyun Zhang XH, Ping Y, Wang J (2012) Establishment of in vitro model of neonatal mouse cortical neuronal injury induced by glutamate. J Neurol Anat 4:363–367
Hernández CC, Burgos CF, Gajardo AH, Silva-Grecchi T, Gavilan J, Toledo JR, Fuentealba J (2017) Neuroprotective effects of erythropoietin on neurodegenerative and ischemic brain diseases: the role of erythropoietin receptor. Neural Regen Res 12:1381
Jeong JE, Park JH, Kim CS, Lee SL, Chung HL, Kim WT, Lee EJ (2017) Neuroprotective effects of erythropoietin against hypoxic injury via modulation of the mitogen-activated protein kinase pathway and apoptosis. Korean J Pediatr 60:181–188. https://doi.org/10.3345/kjp.2017.60.6.181
Jia Y et al (2014) EPO-dependent activation of PI3K/Akt/FoxO3a signalling mediates neuroprotection in in vitro and in vivo models of Parkinson’s disease. J Mol Neurosci 53:117–124. https://doi.org/10.1007/s12031-013-0208-0
Li D-J, Li Y-H, Yuan H-B, Qu L-F, Wang P (2017) The novel exercise-induced hormone irisin protects against neuronal injury via activation of the Akt and ERK1/2 signaling pathways and contributes to the neuroprotection of physical exercise in cerebral ischemia. Metabolism 68:31–42
Malla RR, Asimi R, Teli MA, Shaheen F, Bhat MA (2017) Erythropoietin monotherapy in perinatal asphyxia with moderate to severe encephalopathy: a randomized placebo-controlled trial. J Perinatol 37:596–601. https://doi.org/10.1038/jp.2017.17
Mikrogeorgiou A et al (2017) Dedifferentiated fat cells as a novel source for cell therapy to target neonatal hypoxic-ischemic encephalopathy. Dev Neurosci 39:273–286. https://doi.org/10.1159/000455836
Ostrowski D, Heinrich R (2018) Alternative erythropoietin receptors in the nervous system. J Clin Med 7:24
Salakou S et al (2007) Increased Bax/Bcl-2 ratio up-regulates caspase-3 and increases apoptosis in the thymus of patients with myasthenia gravis. In Vivo 21:123–132
Savoia C, Sisalli MJ, Di Renzo G, Annunziato L, Scorziello A (2011) Rosuvastatin-induced neuroprotection in cortical neurons exposed to OGD/reoxygenation is due to nitric oxide inhibition and ERK1/2 pathway activation. Int J Physiol Pathophysiol Pharmacol 3:57–64
Souvenir R, Flores JJ, Ostrowski RP, Manaenko A, Duris K, Tang J (2014) Erythropoietin inhibits HIF-1alpha expression via upregulation of PHD-2 transcription and translation in an in vitro model of hypoxia-ischemia. Transl Stroke Res 5:118–127. https://doi.org/10.1007/s12975-013-0312-z
Suarez-Mendez S, Tovilla-Zárate CA, Juárez-Rojop IE, Bermúdez-Ocaña DY (2018) Erythropoietin: a potential drug in the management of diabetic neuropathy. Biomed Pharmacother 105:956–961
Tian T, Zeng J, Zhao G, Zhao W, Gao S, Liu L (2018) Neuroprotective effects of orientin on oxygen-glucose deprivation/reperfusion-induced cell injury in primary culture of rat cortical neurons. Exp Biol Med (Maywood NJ) 243:78–86. https://doi.org/10.1177/1535370217737983
Wu YW et al (2016) High-dose erythropoietin and hypothermia for hypoxic-ischemic encephalopathy: a phase II trial. Pediatrics. https://doi.org/10.1542/peds.2016-0191
Xu G, Gong Z, Yu W, Gao L, He S, Qian Z (2007) Increased expression ratio of Bcl-2/Bax is associated with crocin-mediated apoptosis in bovine aortic endothelial cells. Basic Clin Pharmacol Toxicol 100:31–35. https://doi.org/10.1111/j.1742-7843.2007.00001.x
Yan F et al (2016) Erythropoietin improves hypoxic-ischemic encephalopathy in neonatal rats after short-term anoxia by enhancing angiogenesis. Brain Res 1651:104–113. https://doi.org/10.1016/j.brainres.2016.09.024
Yan X et al (2017) Vanillin protects dopaminergic neurons against inflammation-mediated cell death by inhibiting ERK1/2, p38 and the NF-κB signaling pathway. Int J Mol Sci 18:389
Yoo S-J et al (2017) The erythropoietin-derived peptide MK-X and erythropoietin have neuroprotective effects against ischemic brain damage. Cell Death Dis 8:e3003
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This study was sponsored by the Guizhou Science and Technology Cooperation Plan (Qian Ke He LH Zi [2014]7022).
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Si, W., Wang, J., Li, M. et al. Erythropoietin protects neurons from apoptosis via activating PI3K/AKT and inhibiting Erk1/2 signaling pathway. 3 Biotech 9, 131 (2019). https://doi.org/10.1007/s13205-019-1667-y
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DOI: https://doi.org/10.1007/s13205-019-1667-y