Ceruloplasmin (CP) is an essential ferroxidase that is involved in maintaining iron homeostasis by oxidizing toxic ferrous iron (Fe2+) to less-toxic ferric iron (Fe3+). CP has been well studied in many neurodegenerative diseases, but there has not been an in-depth investigation in intracerebral hemorrhage (ICH). This research investigated brain CP expression in rats after ICH and the effect of CP on Fe2+-induced brain injury. This study had two parts: first, rats had injection of autologous blood into the right basal ganglia and the time course of CP expression in the brain examined (protein and mRNA). Second, rats had an injection of either Fe2+ in saline, Fe2+ plus CP in saline, or saline alone into the right basal ganglia. All rats in the second part had T2-weighted magnetic resonance imaging, and behavioral tests before the brains were harvested for immunohistochemistry and Western blotting. We found that CP was expressed on neurons and astrocytes in both cortex and basal ganglia after ICH. The time course showed that ICH induced CP expression increased from 4 h to 7 days, peaking at day 3. Whether the brain itself can produce CP was confirmed by RT-PCR. Exogenous CP reduced Fe2+-induced T2 lesions, blood-brain barrier disruption, brain cell death, and neurological deficits. These results suggest a role of CP in potentially reducing ICH-induced brain injury.
Blood-brain barrier disruption Cerebral hemorrhage Ceruloplasmin Iron Neuronal death
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This work was supported by grants NS-091545, NS-090925, NS-096917, and NS-106746 from the National Institutes of Health (NIH).
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
All institutional and national guidelines for the care and use of laboratory animals were followed.
Xi G, Keep RF, Hoff JT. Mechanisms of brain injury after intracerebral hemorrhage. Lancet Neurol. 2006;5:53–63.CrossRefGoogle Scholar
Wilkinson DA, Pandey AS, Thompson BG, Keep RF, Hua Y, Xi G. Injury mechanisms in acute intracerebral hemorrhage. Neuropharmacology. 2018;134:240–8.CrossRefGoogle Scholar
Garton T, Keep RF, Wilkinson DA, Strahle JM, Hua Y, Garton HJ, et al. Intraventricular hemorrhage: the role of blood components in secondary injury and hydrocephalus. Transl Stroke Res. 2016;7:447–51.CrossRefGoogle Scholar
Xiong XY, Wang J, Qian ZM, Yang QW. Iron and intracerebral hemorrhage: from mechanism to translation. Transl Stroke Res. 2014;5:429–41.CrossRefGoogle Scholar
Bielli P, Calabrese L. Structure to function relationships in ceruloplasmin: a ‘moonlighting’ protein. Cell Mol Life Sci. 2002;59:1413–27.CrossRefGoogle Scholar
Klomp LW, Farhangrazi ZS, Dugan LL, Gitlin JD. Ceruloplasmin gene expression in the murine central nervous system. J Clin Invest. 1996;98:207–15.CrossRefGoogle Scholar
Zheng M, Du H, Ni W, Koch LG, Britton SL, Keep RF, et al. Iron-induced necrotic brain cell death in rats with different aerobic capacity. Transl Stroke Res. 2015;6:215–23.CrossRefGoogle Scholar
Jin H, Xi G, Keep RF, Wu J, Hua Y. Darpp-32 to quantify intracerebral hemorrhage-induced neuronal death in basal ganglia. Transl Stroke Res. 2013;4:130–4.CrossRefGoogle Scholar
Dang G, Yang Y, Wu G, Hua Y, Keep RF, Xi G. Early erythrolysis in the hematoma after experimental intracerebral hemorrhage. Transl Stroke Res. 2017;8:174–82.CrossRefGoogle Scholar
Hua Y, Xi G, Keep RF, Wu J, Jiang Y, Hoff JT. Plasminogen activator inhibitor-1 induction after experimental intracerebral hemorrhage. J Cereb Blood Flow Metab. 2002;22:55–61.CrossRefGoogle Scholar
Ni W, Zheng M, Xi G, Keep RF, Hua Y. Role of lipocalin-2 in brain injury after intracerebral hemorrhage. J Cereb Blood Flow Metab. 2015;35:1454–61.CrossRefGoogle Scholar
Wan S, Cheng Y, Jin H, Guo D, Hua Y, Keep RF, et al. Microglia activation and polarization after intracerebral hemorrhage in mice: the role of protease-activated receptor-1. Transl Stroke Res. 2016;7:478–87.CrossRefGoogle Scholar
Hua Y, Schallert T, Keep RF, Wu J, Hoff JT, Xi G. Behavioral tests after intracerebral hemorrhage in the rat. Stroke. 2002;33:2478–84.CrossRefGoogle Scholar
Altamura C, Squitti R, Pasqualetti P, Gaudino C, Palazzo P, Tibuzzi F, et al. Ceruloplasmin/transferrin system is related to clinical status in acute stroke. Stroke. 2009;40:1282–8.CrossRefGoogle Scholar
Kristinsson J, Snaedal J, Torsdottir G, Johannesson T. Ceruloplasmin and iron in Alzheimer’s disease and Parkinson’s disease: a synopsis of recent studies. Neuropsychiatr Dis Treat. 2012;8:515–21.PubMedPubMedCentralGoogle Scholar
Vassiliev V, Harris ZL, Zatta P. Ceruloplasmin in neurodegenerative diseases. Brain Res Rev. 2005;49:633–40.CrossRefGoogle Scholar
Kaneko K, Hineno A, Yoshida K, Ikeda S. Increased vulnerability to rotenone-induced neurotoxicity in ceruloplasmin-deficient mice. Neurosci Lett. 2008;446:56–8.CrossRefGoogle Scholar
Chang YZ, Qian ZM, Du JR, Zhu L, Xu Y, Li LZ, et al. Ceruloplasmin expression and its role in iron transport in c6 cells. Neurochem Int. 2007;50:726–33.CrossRefGoogle Scholar
Patel BN, Dunn RJ, Jeong SY, Zhu Q, Julien JP, David S. Ceruloplasmin regulates iron levels in the cns and prevents free radical injury. J Neurosci. 2002;22:6578–86.CrossRefGoogle Scholar
Zanardi A, Conti A, Cremonesi M, D'Adamo P, Gilberti E, Apostoli P, et al. Ceruloplasmin replacement therapy ameliorates neurological symptoms in a preclinical model of aceruloplasminemia. EMBO Mol Med. 2018;10:91–106.CrossRefGoogle Scholar
Patel BN, Dunn RJ, David S. Alternative rna splicing generates a glycosylphosphatidylinositol-anchored form of ceruloplasmin in mammalian brain. J Biol Chem. 2000;275:4305–10.CrossRefGoogle Scholar
Yang S, Hua Y, Nakamura T, Keep RF, Xi G. Upregulation of brain ceruloplasmin in thrombin preconditioning. Acta Neurochir Suppl. 2006;96:203–6.CrossRefGoogle Scholar
Zhao F, Xi G, Liu W, Keep RF, Hua Y. Minocycline attenuates iron-induced brain injury. Acta Neurochir Suppl. 2016;121:361–5.CrossRefGoogle Scholar
Gaasch JA, Lockman PR, Geldenhuys WJ, Allen DD, Van der Schyf CJ. Brain iron toxicity: differential responses of astrocytes, neurons, and endothelial cells. Neurochem Res. 2007;32:1196–208.CrossRefGoogle Scholar
Welch KD, Davis TZ, Van Eden ME, Aust SD. Deleterious iron-mediated oxidation of biomolecules. Free Radic Biol Med. 2002;32:577–83.CrossRefGoogle Scholar
Calabrese V, Lodi R, Tonon C, D'Agata V, Sapienza M, Scapagnini G, et al. Oxidative stress, mitochondrial dysfunction and cellular stress response in friedreich’s ataxia. J Neurol Sci. 2005;233:145–62.CrossRefGoogle Scholar
Shamoto-Nagai M, Maruyama W, Yi H, Akao Y, Tribl F, Gerlach M, et al. Neuromelanin induces oxidative stress in mitochondria through release of iron: mechanism behind the inhibition of 26s proteasome. J Neural Transm. 2006;113:633–44.CrossRefGoogle Scholar
Garton T, Keep RF, Hua Y, Xi G. Brain iron overload following intracranial haemorrhage. Stroke Vasc Neurol. 2016;1:172–84.CrossRefGoogle Scholar
Karwacki Z, Kowianski P, Dziewatkowski J, Domaradzka-Pytel B, Ludkiewcz B, Wojcik S, et al. Apoptosis in the course of experimental intracerebral haemorrhage in the rat. Folia Morphol (Warsz). 2005;64:248–52.Google Scholar
David S, Patel BN. Ceruloplasmin: structure and function of an essential ferroxidase. Adv Struct Biol. 2000;6:211–37.CrossRefGoogle Scholar
Zhao L, Hadziahmetovic M, Wang C, Xu X, Song Y, Jinnah HA, et al. Cp/heph mutant mice have iron-induced neurodegeneration diminished by deferiprone. J Neurochem. 2015;135:958–74.CrossRefGoogle Scholar
Texel SJ, Zhang J, Camandola S, Unger EL, Taub DD, Koehler RC, et al. Ceruloplasmin deficiency reduces levels of iron and bdnf in the cortex and striatum of young mice and increases their vulnerability to stroke. PLoS One. 2011;6:e25077.CrossRefGoogle Scholar
Shin EJ, Jeong JH, Chung CK, Kim DJ, Wie MB, Park ES, et al. Ceruloplasmin is an endogenous protectant against kainate neurotoxicity. Free Radic Biol Med. 2015;84:355–72.CrossRefGoogle Scholar