Abstract
In the present study, the effects of erythropoietin (EPO) on preventing adult neurons from apoptosis (introduced by brachial plexus avulsion) were examined, and the mechanism was analyzed. Fifty injury rat models were established in this study by using micro-hemostat forceps to pull out brachial plexus root from the intervertebral foramen in supine position. These models were divided into EPO group (avulsion + 1000 U/kg subcutaneously on alternate days) and control group (avulsion + normal saline). C5-T1 spinal cord was harvested at days 1, 2, 4, 7, and 14. Compared with the control group, the apoptosis of spinal motoneurons was significantly decreased on days 4 and 7 in the EPO group, which was also approved by TUNEL examination results. The detection of p-JNK and expression of c-Jun and cleavage of cleaved PARP (c-PARP) were also examined by immunohistochemistry and were increased immediately at day 1, and peaked at day 2, day 2, and day 4 in control group, respectively. However, the amounts were decreased and delayed by EPO treatment significantly at the same time points. In conclusion, the apoptosis of adult spinal motorneurons was associated with JNK phosphorylation, c-Jun expression, and caspase activity, and EPO-mediated neuronal protective effect is proved by downregulating the JNK phosphorylation and c-Jun expression and inhibiting of c-PARP cleavage.
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Behrens A, Sibilia M, Wagner EF (1999) Amino-terminal phosphorylation of c-Jun regulates stress-induced apoptosis and cellular proliferation. Nat Genet 21:326–329
Bergerot A, Shortland PJ, Anand P, Hunt SP, Carlstedt T (2004) Co-treatment with riluzole and GDNF is necessary for functional recovery after ventral root avulsion injury. Exp Neurol 187:359–366
Bernaudin M, Nedelec AS, Divoux D (2002) Normobaric hypoxia induces tolerance to focal permanent cerebral ischemia in association with an increased expression of hypoxia-inducible factor-1 and its target genes, erythropoietin and VEGF, in the adult mouse brain. J Cereb Blood Flow Metab 22:393–403
Bertelli JA, Ghizoni MF (2010) Nerve root grafting and distal nerve transfern for C5-C6 brachial plexus injuries. J Hand Surg [Am] 35:769–775
Celik M, Gokmen N, Erbayraktar S (2002) Erythropoietin prevents motorneuron apoptosis and neurologic disability in experimental spinal cord ischemic injury. Proc Natl Acad Sci U S A 99:2258–2263
Celilk M, Gokmen N, Erbayraktar S, Akhisaroglu M, Konakc S, Ulukus C, Genc S, Genc K, Sagiroglu E, Cerami A, Brines M (2002) Erythropoietin prevents motor neuron apoptosis and neurologic disability in experimental spinal cord ischemic injury. Proc Natl Acad Sci U S A 99:2258–2263
Chen ZY, Chai YF, Cao L, Lu CL, He C (2001) Glial cell line-derived neurotrophic factor enhances axonal regeneration following sciatic nerve transection in adult rats. Brain Res 902:272–276
Chikuma M (2000) Tissue-specific regulation of erythropoietin production in the murine kidney, brain and uterus. Am J Physiol 279:E1242–E1248
Dame C, Bartmann P, Wolber EM (2000) Erythropoietin gene expression in different areas of the developing human central nervous system. Dev Brain Res 125:69–74
Davis RJ (2000) Signal transduction by the JNK group of MAP kinases. Cell 103:239–252
Digicaylioglu M, Lipton SA (2001) Erythropoietin-mediated neuroprotection involves cross-talk between Jak2 and NF-kappa B signaling cascades. Nature 412:641–647
Fan M, Goodwin M, Vi T, Brantley-Finley C, Gaarde WA, Chambers TC (2000) Vinblastine-induced phosphorylation of Bcl-2 and Bcl-XL is mediated by JNK and occurs in parallel with inactivation of the Raf-1/MEK/ERK cascade. J Biol Chem 275:29980–29985
Fox IK, Brenner MJ, Johnson PJ, Hunter DA, Mackinnon SE (2012) Axonal regeneration and motor neuron survival after microsurgical nerve reconstruction. Microsurgery 32:552–562
Gao Y, Signore AP, Yin W, Cao G, Yin XM, Sun F, Luo Y, Graham SH, Chen J (2005) Neuroprotection against focal ischemic brain injury by inhibition of c-Jun N-terminal kinase and attenuation of the mitochondria) apoptosis-signaling pathway. J Cereb Blood Flow Metab 25:694–712
Gorio A, Gokmen N, Erbayraktar S, Yilmaz O, Madaschi Z, Cichetti C, Giulio MD, Vardar E, Cerami A, Brines M (2002) Recombinant human erythropoietin counteracts secondary injury and markedly enhances neurological recovery from experimental spinal cord trauma. Proc Natl Acad Sci U S A 99:9450–9455
Grasso G, Sfacteria A, Erbayraktar S (2006) Amelioration of spinal cord compressive injury by pharmacological preconditioning with erythropoietin and a nonerythropoietic erythropoietin derivative. J Neurosurg Spine 4:310–318
Gu Y, Spasic Z, Wu W (1997) The effects of remaining axons on motoneuron survival and NOS expression following axotomy in the adult rat. Dev Neurosci 19:255–259
Gu HY, Chai H, Zhang ZB, Yao LH, Zhou LH, Wong WM, Bruce IC, Wu WT (2005) Survival, regeneration and functional recovery of motoneurons after delayed reimplantation of avulsed spinal root in adult rat. Exp Neurol 192:89–99
Hess TC, Yang P, Xu DD, Turner J, Nimnual TK, Barsagi A, Hones D, Flavell SN, Davis RA (2000) Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. Science 288:870–874
Ho FM, Liu SH, Liau CS, Huang PJ, Lin-Shiau SY (2000) High glucose-induced apoptosis in human endothelial cells is mediated by sequential activations of c-Jun NH(2)-terminal kinase and caspase-3. Circulation 101:2618–2624
Houle JD, Schramm P, Herdeqen T (1998) Trophic factor modulation of c-Jun expression in supraspinal neurons after chronic spinal cord injury. Exp Neurol 154:602–611
Ichikawa H, Kano M, Shimizu Y, Suzuki T, Sawada E, Ono W, Chu LW, Cote PD (2010) Increase of c-Fos and c-Jun expression in spinal and cranial motoneurons ofthe degenerating muscle mouse (ScnBa(dmu)). Cell Mol Neurobiol 30:737–742
Jerome JT, Rajmohan B (2012) Axillary nerve neurotization with the anterior deltopectoral approach in brachial plexus injuries. Microsurgery 32:445–451
Konishi Y, Chui DH, Hirose H, Kunishita T, Tabira T (1993) Trophic effect of erythropoietin and other hematopoietic factors on central cholinergic neurons in vitro and in vivo. Brain Res 609:29–35
Kretz A, Happold CJ, Marticke JK, Isenmann S (2005) Erythropoietin promotes regeneration of adult CNS neurons via Jak2/Stat3 and PI3K/AKT pathway activation. Mol Cell Neurosci 29:569–579
Le-Niculescu H, Bonfoco E, Kasuya Y, Claret FX, Green DR, Karin M (1999) Withdrawal of survival factors results in activation of the JNK pathway in neuronal cells leading to Fas ligand induction and cell death. Mol Cell Biol 19:751–763
Li L, Wu W, Lin LF, Lei M, Oppenheim RW, Houenou LJ (1995) Rescue of adult mouse motoneurons from injury-induced cell death by filial cell line-derived neurotrophic factor. Proc Natl Acad Sci U S A 92:9771–9775
McKay Hart A, Brannstrom T, Wiberg M, Terenghi G (2002) Primary sensory neurons and satellite cells after peripheral axotomy in the adult rat-timecourse of cell death and elimination. Exp Brain Res 142:308–318
Novikov L, Novikova L, Kellerth JO (1995) Brain-derived neurotrophic factor promotes survival and blocks nitric oxide synthase expression in adult rat spinal motoneurons after ventral root avulsion. Neurosci Lett 200:45–48
Okuno S, Saito A, Hayashi T, Chan PH (2004) The c-Jun N-terminal protein kinase signaling pathway mediates Bax activation and subsequent neuronal apoptosis through interaction with Bim after transient focal cerebral ischemia. J Neurosci 24:7879–7887
Oliveira AL, Riding M, Negro A, Langone F, Cullheim S (2002) Apoptosis of spinal Interneurons induced by sciatic nerve axotomy in the neonatal rat is counteracted by nerve growth factor and ciliary neurotrophic factor. J Comp Neurol 447:381–393
Pardridge WM (1998) CNS drug design based on principles of blood-brain barrier transport. J Neurochem 70:1781–1792
Raivich G, Bohatschek M, Da Costa C, Iwata O, Galiano M, Hristova M, Nateri AS, Makwana M, Riera-Sans L, Wolfer DP, Lipp HP, Aguzzi A, Wagner EF, Behrens A (2004) The AP-1 transcription factor c-Jun is required for efficient axonal regeneration. Neuron 43:57–67
Sabapathy K, Hu Y, Kallunki T, Schreiber M, David JP, Jochum W, Wagner EF, Karin M (1999) JNK2 is required for efficient T-cell activation and apoptosis but not for normal lymphocyte development. Curr Biol 9:116–125
Sinor DA, Greenberg AD (2000) Erythropoietin protects cultured cortical neurons, but not astroglia, from hypoxia and AMPA toxicity. Neurosci Lett 290:213–215
Siren AL, Fratelli M, Brines M (2001) Erythropoietin prevents neuronal apoptosis after cerebral ischemia and metabolic stress. Proc Natl Acad Sci U S A 98:4044–4049
Tang Z, Sun X, Huo G, Xie Y, Shi Q, Chen S, Wang X, Liao Z (2013) Protective effects of erythropoietin on astrocytic swelling after oxygen-glucose deprivation and reoxygenation: mediation through AQP4 expression and MAPK pathway. Neuropharmacology 67:8–15
Thibault K, Rivals I, M’Dahoma S, Dubacq S, Pezet S, Calvino B (2013) Structural and molecular alterations of primary afferent fibres in the spinal dorsal Horn in Vincristine-induced neuropathy in rat. J Mol Neurosci 51:880–892
Tournier C, Hess P, Yang DD, Xu J, Turner TK, Nimnual A, BarSagi D, Jones SN, Flavell RA, Davis RJ (2000) Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway. Science 288:870–874
Wang S, Yiu HW, Li Y, Wang H, Pan Y (2012) Contralateral C7 nerve root transfer to neurotize the upper trunk via a modified prespinal route in repair of brachial plexus avulsion injury. Microsurgery 32:183–188
Wu D, Li Q, Zhu X, Wu G, Cui S (2013) Valproic acid protection againse the brachial plexus root avulsion-induced death of motoneurons in rats. Microsurgery 13:2077–2085
Wu Q, Jiang Y, Yuan X, Zhang X, Li B, Liu M, Wang B, Li H, Liu S, Xiu R (2014) Melatonin treatment protects against acute spinal cord injury-induced disruption of blood spinal cord barrier in mice. J Mol Neurosci 54:714–722
Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME (2008) Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science 270:1326–1331
Yang DD, Kuan CY, Whitmarsh AJ, Rincon M, Zheng TS, Davis RJ, Rakic P, Flavell RA (1997) Absence of excitotoxicity-induced apoptosis in the hippocampus of mice lacking the Jnk3 gene. Nature 389:865–870
Yin KJ, Kim GM, Lee JM, He YY, Xu J, Hsu CY (2005) JNK activation contributes to DP5 induction and apoptosis following traumatic spinal cord injury. Neurobiol Dis 20:881–889
Yuan Q, Hu B, Wu Y, Chu TH, Su H, Zhang W, So KF, Lin Z, Wu W (2010) Induction of c-Jun phosphorylation in spinal motoneurons in neonatal and aduR rats following axonal injury. Brain Res 1320:7–15
Zhu N, Zhang C, Li Z, Meng Y, Feng B, Wang X, Yang M, Wan L, Ning B, Li S (2014) Experimental study on the effect of electrostimulation on neural regeneration after oculomotor nerve injury. J Mol Neurosci 54:639–652
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Li, K., Cao, RJ., Zhu, XJ. et al. Erythropoietin Attenuates the Apoptosis of Adult Neurons After Brachial Plexus Root Avulsion by Downregulating JNK Phosphorylation and c-Jun Expression and Inhibiting c-PARP Cleavage. J Mol Neurosci 56, 917–925 (2015). https://doi.org/10.1007/s12031-015-0543-4
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DOI: https://doi.org/10.1007/s12031-015-0543-4