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The Role of Neuroglobin in the Neuroprotection of Limb Ischemic Preconditioning in Rats

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Abstract

Recent evidence suggests that limb ischemic preconditioning (LIP) protects neurons against cerebral ischemia-reperfusion injury. However, the mechanisms of LIP are not well understood. Neuroglobin (Ngb) is a recently discovered globin that affords protection against hypoxic/ischemic brain injury. This study was performed to investigate the role of Ngb in the neuroprotection of LIP against brain ischemia and the involvements of mitochondria in the process. The rat global brain ischemic model was used, and the CA1 hippocampus was selected as the observational target. Ngb expression was investigated by RT-PCR and Western blot. Neuropathological evaluation was performed by thionin staining. Mitochondrial membrane potential (Δψm), Na+-K+-ATPase activity, and ultrastructure were examined by flow cytometry, spectrophotometry, and transmission electron microscopy, respectively. We also used Ngb antisense oligodeoxynucleotides (AS-ODNs) and Ngb inducer hemin to inhibit or mimic the effect of LIP. We found that LIP significantly up-regulated Ngb expression and protected neurons against ischemia. Furthermore, LIP effectively improved deterioration in the Δψm, mitochondrial Na+-K+-ATPase activity, and ultrastructure induced by cerebral ischemia. These effects of LIP were inhibited partly by Ngb AS-ODNs and mimicked by hemin. It could be concluded that up-regulation of Ngb expression played an important role in the neuroprotection induced by LIP, and the Ngb-mediated neuroprotection of LIP was, at least partly, associated with mitochondria.

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References

  1. Abu-Amara M, Yang SY, Quaglia A, Rowley P, Tapuria N, Fuller B, Davidson B, Seifalian A (2012) The hepatic soluble guanylyl cyclase-cyclic guanosine monophosphate pathway mediates the protection of remote ischemic preconditioning on the microcirculation in liver ischemia-reperfusion injury. Transplantation 93:880–886

    Article  PubMed  CAS  Google Scholar 

  2. Koch S, Katsnelson M, Dong C, Perez-Pinzon M (2011) Remote ischemic limb preconditioning after subarachnoid hemorrhage: a phase Ib study of safety and feasibility. Stroke 42:1387–1391

    Article  PubMed  Google Scholar 

  3. Tapuria N, Junnarkar S, Abu-Amara M, Fuller B, Seifalian AM, Davidson BR (2012) Modulation of microcirculatory changes in the late phase of hepatic ischaemia-reperfusion injury by remote ischaemic preconditioning. HPB (Oxford) 14:87–97

    Article  Google Scholar 

  4. Sun XC, Xian XH, Li WB, Li L, Yan CZ, Li QJ, Zhang M (2010) Activation of p38 MAPK participates in brain ischemic tolerance induced by limb ischemic preconditioning by up-regulating HSP 70. Exp Neurol 224:347–355

    Article  PubMed  CAS  Google Scholar 

  5. Burmester T, Weich B, Reinhardt S, Hankeln T (2000) A vertebrate globin expressed in the brain. Nature 407:520–523

    Article  PubMed  CAS  Google Scholar 

  6. Hota KB, Hota SK, Srivastava RB, Singh SB (2012) Neuroglobin regulates hypoxic response of neuronal cells through Hif-1α- and Nrf2-mediated mechanism. J Cereb Blood Flow Metab 32:1046–1060

    Article  PubMed  CAS  Google Scholar 

  7. Raychaudhuri S, Skommer J, Henty K, Birch N, Brittain T (2010) Neuroglobin protects nerve cells from apoptosis by inhibiting the intrinsic pathway of cell death. Apoptosis 15:401–411

    Article  PubMed  CAS  Google Scholar 

  8. Liu J, Yu Z, Guo S, Lee SR, Xing C, Zhang C, Gao Y, Nicholls DG, Lo EH, Wang X (2009) Effects of neuroglobin overexpression on mitochondrial function and oxidative stress following hypoxia/reoxygenation in cultured neurons. J Neurosci Res 87:164–170

    Article  PubMed  CAS  Google Scholar 

  9. Liu N, Yu Z, Xiang S, Zhao S, Tjärnlund-Wolf A, Xing C, Zhang J, Wang X (2012) Transcriptional regulation mechanisms of hypoxia-induced neuroglobin gene expression. Biochem J 443:153–164

    Article  PubMed  CAS  Google Scholar 

  10. Zhao S, Yu Z, Zhao G, Xing C, Hayakawa K, Whalen MJ, Lok JM, Lo EH, Wang X (2012) Neuroglobin-overexpression reduces traumatic brain lesion size in mice. BMC Neurosci 13:67

    Article  PubMed  CAS  Google Scholar 

  11. Sun Y, Jin K, Peel A, Mao XO, Xie L, Greenberg DA (2003) Neuroglobin protects the brain from experimental stroke in vivo. Proc Natl Acad Sci U S A 100:3497–3500

    Article  PubMed  CAS  Google Scholar 

  12. Wang X, Liu J, Zhu H, Tejima E, Tsuji K, Murata Y, Atochin DN, Huang PL, Zhang C, Lo EH (2008) Effects of neuroglobin overexpression on acute brain injury and long-term outcomes after focal cerebral ischemia. Stroke 39:1869–1874

    Article  PubMed  CAS  Google Scholar 

  13. Vosler PS, Graham SH, Wechsler LR, Chen J (2009) Mitochondrial targets for stroke: focusing basic science research toward development of clinically translatable therapeutics. Stroke 40:3149–3155

    Article  PubMed  CAS  Google Scholar 

  14. Correia SC, Cardoso S, Santos RX, Carvalho C, Santos MS, Perry G, Smith MA, Moreira PI (2011) New insights into the mechanisms of mitochondrial preconditioning-triggered neuroprotection. Curr Pharm Des 17:3381–3389

    Article  PubMed  CAS  Google Scholar 

  15. Dirnagl U, Meisel A (2008) Endogenous neuroprotection: mitochondria as gateways to cerebral preconditioning? Neuropharmacology 55:334–344

    Article  PubMed  CAS  Google Scholar 

  16. Stanika RI, Winters CA, Pivovarova NB, Andrews SB (2010) Differential NMDA receptor-dependent calcium loading and mitochondrial dysfunction in CA1 vs. CA3 hippocampal neurons. Neurobiol Dis 37:403–411

    Article  PubMed  CAS  Google Scholar 

  17. Zhang B, Wei X, Cui X, Kobayashi T, Li W (2008) Effects of heme oxygenase 1 on brain edema and neurologic outcome after cardiopulmonary resuscitation in rats. Anesthesiology 109:260–268

    Article  PubMed  CAS  Google Scholar 

  18. Robin E, Simerabet M, Hassoun SM, Adamczyk S, Tavernier B, Vallet B, Bordet R, Lebuffe G (2011) Postconditioning in focal cerebral ischemia: role of the mitochondrial ATP-dependent potassium channel. Brain Res 1375:137–146

    Article  PubMed  CAS  Google Scholar 

  19. Oselkin M, Tian D, Bergold PJ (2010) Low-dose cardiotonic steroids increase sodium-potassium ATPase activity that protects hippocampal slice cultures from experimental ischemia. Neurosci Lett 473:67–71

    Article  PubMed  CAS  Google Scholar 

  20. Liu YX, Zhang M, Liu LZ, Cui X, Hu YY, Li WB (2012) The role of glutamate transporter-1a in the induction of brain ischemic tolerance in rats. Glia 60:112–124

    Article  PubMed  Google Scholar 

  21. Pulsinelli WA, Brierley JB (1979) A new model of bilateral hemispheric ischemia in the unanesthetized rat. Stroke 10:267–272

    Article  PubMed  CAS  Google Scholar 

  22. Hundahl C, Kelsen J, Kjaer K, Rønn LC, Weber RE, Geuens E, Hay-Schmidt A, Nyengaard JR (2006) Does neuroglobin protect neurons from ischemic insult? A quantitative investigation of neuroglobin expression following transient MCAo in spontaneously hypertensive rats. Brain Res 1085:19–27

    Article  PubMed  CAS  Google Scholar 

  23. Zhang M, Li WB, Geng JX, Li QJ, Sun XC, Xian XH, Qi J, Li SQ (2007) The upregulation of glial glutamate transporter-1 participates in the induction of brain ischemic tolerance in rats. J Cereb Blood Flow Metab 27:1352–1368

    Article  PubMed  CAS  Google Scholar 

  24. Zhang WH, Wang H, Wang X, Narayanan MV, Stavrovskaya IG, Kristal BS, Friedlander RM (2008) Nortriptyline protects mitochondria and reduces cerebral ischemia/hypoxia injury. Stroke 39:455–462

    Article  PubMed  Google Scholar 

  25. Rao S, Lin Z, Drobyshevsky A, Chen L, Ji X, Ji H, Yang Y, Yu L, Derrick M, Silverman RB, Tan S (2011) Involvement of neuronal nitric oxide synthase in ongoing fetal brain injury following near-term rabbit hypoxia-ischemia. Dev Neurosci 33:288–298

    Article  PubMed  CAS  Google Scholar 

  26. Bao CL, Dong HS (2008) Effect of scalp-acupuncture on energy metabolism of brain tissue in intracerebral hemorrhage rats. Zhen Ci Yan Jiu 33:93–97

    PubMed  Google Scholar 

  27. Kelsen J, Hundahl CA, Hay-Schmidt A (2008) Neuroglobin: endogenous neuroprotectant or maintenance of homeostasis? Stroke 39:e177–178

    Article  PubMed  CAS  Google Scholar 

  28. Skommer J, Brittain T (2012) Extended survival of SH-SY5Y cells following overexpression of Lys67Glu neuroglobin is associated with stabilization of Δψ(M). Cytometry A 81:602–610

    PubMed  Google Scholar 

  29. Greenberg DA, Jin K, Khan AA (2008) Neuroglobin: an endogenous neuroprotectant. Curr Opin Pharmacol 8:20–24

    Article  PubMed  CAS  Google Scholar 

  30. Li RC, Guo SZ, Lee SK, Gozal D (2010) Neuroglobin protects neurons against oxidative stress in global ischemia. J Cereb Blood Flow Metab 30:1874–1882

    Article  PubMed  CAS  Google Scholar 

  31. Jin K, Mao Y, Mao X, Xie L, Greenberg DA (2010) Neuroglobin expression in ischemic stroke. Stroke 41:557–559

    Article  PubMed  CAS  Google Scholar 

  32. Shang A, Zhou D, Wang L, Gao Y, Fan M, Wang X, Zhou R, Zhang C (2006) Increased neuroglobin levels in the cerebral cortex and serum after ischemia-reperfusion insults. Brain Res 1078:219–226

    Article  PubMed  CAS  Google Scholar 

  33. Avivi A, Gerlach F, Joel A, Reuss S, Burmester T, Nevo E, Hankeln T (2010) Neuroglobin, cytoglobin, and myoglobin contribute to hypoxia adaptation of the subterranean mole rat Spalax. Proc Natl Acad Sci U S A 107:21570–21575

    Article  PubMed  CAS  Google Scholar 

  34. Khan AA, Wang Y, Sun Y, Mao XO, Xie L, Miles E, Graboski J, Chen S, Ellerby LM, Jin K, Greenberg DA (2006) Neuroglobin-overexpressing transgenic mice are resistant to cerebral and myocardial ischemia. Proc Natl Acad Sci U S A 103:17944–17948

    Article  PubMed  CAS  Google Scholar 

  35. Gao Y, Mengana Y, Cruz YR, Muñoz A, Testé IS, García JD, Wu Y, Rodríguez JC, Zhang C (2011) Different expression patterns of Ngb and EPOR in the cerebral cortex and hippocampus revealed distinctive therapeutic effects of intranasal delivery of Neuro-EPO for ischemic insults to the gerbil brain. J Histochem Cytochem 59:214–227

    Article  PubMed  CAS  Google Scholar 

  36. Shao G, Gong KR, Li J, Xu XJ, Gao CY, Zeng XZ, Lu GW, Huo X (2009) Antihypoxic effects of neuroglobin in hypoxia-preconditioned mice and SH-SY5Y cells. Neurosignals 17:196–202

    Article  PubMed  CAS  Google Scholar 

  37. Sun Y, Jin K, Mao XO, Zhu Y, Greenberg DA (2001) Neuroglobin is up-regulated by and protects neurons from hypoxic-ischemic injury. Proc Natl Acad Sci U S A 98:15306–15311

    Article  PubMed  CAS  Google Scholar 

  38. Qi D, Liu H, Niu J, Fan X, Wen X, Du Y, Mou J, Pei D, Liu Z, Zong Z, Wei X, Song Y (2012) Heat shock protein 72 inhibits c-Jun N-terminal kinase 3 signaling pathway via Akt1 during cerebral ischemia. J Neurol Sci 317:123–129

    Article  PubMed  CAS  Google Scholar 

  39. He AW, Yang T, Chen SQ, Li ZP, Li HP, Huang WJ, Cheng JY, Zhang J, Yang P, Wang WT (2011) Effects of hemin on neuroglobin expression after cardiopulmonary resuscitation in rats. World J Emerg Med 2:54–58

    Article  Google Scholar 

  40. Zhu Y, Sun Y, Jin K, Greenberg DA (2002) Hemin induces neuroglobin expression in neural cells. Blood 100:2494–2498

    Article  PubMed  CAS  Google Scholar 

  41. Lim SY, Hausenloy DJ (2012) Remote ischemic conditioning: from bench to bedside. Front Physiol 3:27

    Article  PubMed  Google Scholar 

  42. Veighey K, Macallister RJ (2012) Clinical applications of remote ischemic preconditioning. Cardiol Res Pract 2012:620681

    PubMed  Google Scholar 

  43. Haines B, Demaria M, Mao X, Xie L, Campisi J, Jin K, Greenberg DA (2012) Hypoxia-inducible factor-1 and neuroglobin expression. Neurosci Lett 514:137–140

    Article  PubMed  CAS  Google Scholar 

  44. Yu Z, Xu J, Liu N, Wang Y, Li X, Pallast S, van Leyen K, Wang X (2012) Mitochondrial distribution of neuroglobin and its response to oxygen-glucose deprivation in primary-cultured mouse cortical neurons. Neuroscience 218:235–242

    Article  PubMed  CAS  Google Scholar 

  45. Antao ST, Duong TT, Aran R, Witting PK (2010) Neuroglobin overexpression in cultured human neuronal cells protects against hydrogen peroxide insult via activating phosphoinositide-3 kinase and opening the mitochondrial K(ATP) channel. Antioxid Redox Signal 13:769–781

    Article  PubMed  CAS  Google Scholar 

  46. Xu WL, Wang CL, Liao ZY, Zhang YL, Yu LH, Meng FW, Wang XX, Yin ZY, Qian LJ, Zhang CG (2003) Identification of interaction and interaction domains between neuroglobin and Na+, K+-ATPase beta2 subunit. Acta Biochim Biophys Sin 35:823–828

    PubMed  CAS  Google Scholar 

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Acknowledgments

This study was supported by National Natural Science Foundation of China (No: 30770738, 81000477, 81271454), the Key Base Research Project of Hebei Province (No: 11966121D), and Natural Science Foundation of Hebei Province (No: C2010000460, C2010000478, C2011206039).

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Authors and Affiliations

  1. Department of Pathophysiology, Hebei Medical University, 361 Zhongshan East Road, Shijiazhuang, 050017, China

    Shu-Qin Li, Wen-Bin Li, Min Zhang & Yu-Yan Hu

  2. Department of Cardiology, Second Hospital of Hebei Medical University, Shijiazhuang, 050000, China

    Yu-Zhou Wu

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  1. Shu-Qin Li
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Correspondence to Wen-Bin Li.

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Li, SQ., Li, WB., Zhang, M. et al. The Role of Neuroglobin in the Neuroprotection of Limb Ischemic Preconditioning in Rats. Mol Neurobiol 47, 197–208 (2013). https://doi.org/10.1007/s12035-012-8373-7

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  • DOI: https://doi.org/10.1007/s12035-012-8373-7

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