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Cited by 33Neuroinflammation and Neurodegeneration MechanismsInflammasome and immune disordersNeurological Disease Mechanisms and Treatments

Ceruloplasmin potentiates nitric oxide synthase activity and cytokine secretion in activated microglia

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Lazzaro, Massimo, Bettegazzi, Barbara, Barbariga, Marco, Codazzi, Franca, Zacchetti, Daniele, Alessio, Massimo·2014·Journal of Neuroinflammation
DOI 10.1186/s12974-014-0164-9PMC PMC4174266
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Abstract

BACKGROUND: Ceruloplasmin is a ferroxidase expressed in the central nervous system both as soluble form in the cerebrospinal fluid (CSF) and as membrane-bound GPI-anchored isoform on astrocytes, where it plays a role in iron homeostasis and antioxidant defense. It has been proposed that ceruloplasmin is also able to activate microglial cells with ensuing nitric oxide (NO) production, thereby contributing to neuroinflammatory conditions. In light of the possible role of ceruloplasmin in neurodegenerative diseases, we were prompted to investigate how this protein could contribute to microglial activation in either its native form, as well as in its oxidized form, recently found generated in the CSF of patients with Parkinson's and Alzheimer's diseases. METHODS: Primary rat microglial-enriched cultures were treated with either ceruloplasmin or oxidized-ceruloplasmin, alone or in combination with lipopolysaccharide (LPS). Production of NO and expression of inducible nitric oxide synthase (iNOS) were evaluated by Griess assay and Western blot analysis, respectively. The productions of the pro-inflammatory cytokine IL-6 and the chemokine MIP-1α were assessed by quantitative RT-PCR and ELISA. RESULTS: Regardless of its oxidative status, ceruloplasmin by itself was not able to activate primary rat microglia. However, ceruloplasmin reinforced the LPS-induced microglial activation, promoting an increase of NO production, as well as the induction of IL-6 and MIP-1α. Interestingly, the ceruloplasmin-mediated effects were observed in the absence of an additional induction of iNOS expression. The evaluation of iNOS activity in primary glial cultures and in vitro suggested that the increased NO production induced by the combined LPS and ceruloplasmin treatment is mediated by a potentiation of the enzymatic activity. CONCLUSIONS: Ceruloplasmin potentiates iNOS activity in microglial cells activated by a pro-inflammatory stimulus, without affecting iNOS expression levels. This action might be mediated by the activation of a yet unknown Cp receptor that triggers intracellular signaling that cross-talks with the response elicited by LPS or other pro-inflammatory stimuli. Therefore, ceruloplasmin might contribute to pathological conditions in the central nervous system by exacerbating neuroinflammation.

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  • GPI-anchored proteins can partition to the cerebrospinal fluid (CSF).

    “GPI-anchored proteins can partition to the cerebrospinal fluid (CSF) (52–54).”

    From GDE2-RECK controls ADAM10 α-secretase–mediated cleavage of amyloid precursor protein (2021) · cites this paper as [54]

    The Background (page unavailable) says ceruloplasmin is released into CSF and that a GPI-anchored form is present on astrocytes and leptomeningeal cells. It does not say that the GPI-anchored form, or GPI-anchored proteins generally, partition into CSF. (LLM, conf=0.82)

    Experimental — accuracy not yet measured

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  1. [1]Hellman NE Gitlin JD Ceruloplasmin metabolism and function Annu Rev Nutr 2002 22 439 458 10.1146/annurev.nutr.22.012502.114457 12055353 · DOI
  2. [2]Patel BN David S A novel glycosylphosphatidylinositol-anchored form of ceruloplasmin is expressed by mammalian astrocytes J Biol Chem 1997 272 20185 20190 10.1074/jbc.272.32.20185 9242695 · DOI · Paper page
  3. [3]Mittal B Doroudchi MM Jeong SY Patel BN David S Expression of a membrane-bound form of the ferroxidase ceruloplasmin by leptomeningeal cells Glia 2003 41 337 346 10.1002/glia.10158 12555201 · DOI
  4. [4]Duce JA Tsatsanis A Cater MA James SA Robb E Wikhe K Leong SL Perez K Johanssen T Greenough MA Cho HH Galatis D Moir RD Masters CL McLean C Tanzi RE Cappai R Barnham KJ Ciccotosto GD Rogers JT Bush AI Iron-export ferroxidase activity of beta-amyloid precursor protein is inhibited by zinc in Alzheimer’s disease Cell 2010 142 857 867 10.1016/j.cell.2010.08.014 20817278 PMC2943017 · DOI
  5. [5]Jeong SY David S Glycosylphosphatidylinositol-anchored ceruloplasmin is required for iron efflux from cells in the central nervous system J Biol Chem 2003 278 27144 27148 10.1074/jbc.M301988200 12743117 · DOI · Paper page
  6. [6]Olivieri S Conti A Iannaccone S Cannistraci CV Campanella A Barbariga M Codazzi F Pelizzoni I Magnani G Pesca M Franciotta D Cappa SF Alessio M Ceruloplasmin oxidation, a feature of Parkinson’s disease CSF, inhibits ferroxidase activity and promotes cellular iron retention J Neurosci 2011 31 18568 18577 10.1523/JNEUROSCI.3768-11.2011 22171055 PMC6623910 · DOI · Paper page
  7. [7]Grimm S Hoehn A Davies KJ Grune T Protein oxidative modifications in the ageing brain: consequence for the onset of neurodegenerative disease Free Radic Res 2011 45 73 88 10.3109/10715762.2010.512040 20815785 PMC3675897 · DOI
  8. [8]Zecca L Youdim MB Riederer P Connor JR Crichton RR Iron, brain ageing and neurodegenerative disorders Nat Rev Neurosci 2004 5 863 873 10.1038/nrn1537 15496864 · DOI · Paper page
  9. [9]Corti A Curnis F Isoaspartate-dependent molecular switches for integrin-ligand recognition J Cell Sci 2011 124 515 522 10.1242/jcs.077172 21282473 · DOI
  10. [10]Barbariga M Curnis F Spitaleri A Andolfo A Zucchelli C Lazzaro M Magnani G Musco G Corti A Alessio M Oxidation-induced structural changes of ceruloplasmin foster NGR-motifs deamidation that promote integrin binding and signalling J Biol Chem 2014 289 3736 3748 10.1074/jbc.M113.520981 24366863 PMC3916571 · DOI
  11. [12]Graeber MB Streit WJ Microglia: biology and pathology Acta Neuropathol 2010 119 89 105 10.1007/s00401-009-0622-0 20012873 · DOI
  12. [14]Akiyama H Barger S Barnum S Bradt B Bauer J Cole GM Cooper NR Eikelenboom P Emmerling M Fiebich BL Finch CE Frautschy S Griffin WS Hampel H Hull M Landreth G Lue L Mrak R Mackenzie IR McGeer PL O’Banion MK Pachter J Pasinetti G Plata-Salaman C Rogers J Rydel R Shen Y Streit W Strohmeyer R Tooyoma I Inflammation and Alzheimer’s disease Neurobiol Aging 2000 21 383 421 10.1016/S0197-4580(00)00124-X 10858586 PMC3887148 · DOI · Paper page
  13. [15]Cameron B Landreth GE Inflammation, microglia, and Alzheimer’s disease Neurobiol Dis 2010 37 503 509 10.1016/j.nbd.2009.10.006 19833208 PMC2823849 · DOI
  14. [16]Teismann P Tieu K Cohen O Choi DK Wu DC Marks D Vila M Jackson-Lewis V Przedborski S Pathogenic role of glial cells in Parkinson’s disease Mov Disord 2003 18 121 129 10.1002/mds.10332 12539204 · DOI
  15. [17]Tufekci KU Meuwissen R Genc S Genc K Inflammation in Parkinson’s disease Adv Protein Chem Struct Biol 2012 88 69 132 10.1016/B978-0-12-398314-5.00004-0 22814707 · DOI
  16. [18]Perry VH Innate inflammation in Parkinson’s disease Cold Spring Harb Perspect Med 2012 2 a009373 10.1101/cshperspect.a009373 22951445 PMC3426823 · DOI
  17. [19]Peterson LJ Flood PM Oxidative stress and microglial cells in Parkinson’s disease Mediators Inflamm 2012 2012 401264 10.1155/2012/401264 22544998 PMC3321615 · DOI
  18. [20]Ricci S Fuso A Ippoliti F Businaro R Stress-induced cytokines and neuronal dysfunction in Alzheimer’s disease J Alzheimers Dis 2012 28 11 24 22124029
  19. [21]Swardfager W Lanctot K Rothenburg L Wong A Cappell J Herrmann N A meta-analysis of cytokines in Alzheimer’s disease Biol Psychiatry 2010 68 930 941 10.1016/j.biopsych.2010.06.012 20692646 · DOI · Paper page
  20. [22]Glass CK Saijo K Winner B Marchetto MC Gage FH Mechanisms underlying inflammation in neurodegeneration Cell 2010 140 918 934 10.1016/j.cell.2010.02.016 20303880 PMC2873093 · DOI · Paper page
  21. [23]Cunningham C Microglia and neurodegeneration: the role of systemic inflammation Glia 2013 61 71 90 10.1002/glia.22350 22674585 · DOI · Paper page
  22. [24]Lehnardt S Massillon L Follett P Jensen FE Ratan R Rosenberg PA Volpe JJ Vartanian T Activation of innate immunity in the CNS triggers neurodegeneration through a Toll-like receptor 4-dependent pathway Proc Natl Acad Sci U S A 2003 100 8514 8519 10.1073/pnas.1432609100 12824464 PMC166260 · DOI · Paper page
  23. [25]Beraud D Hathaway HA Trecki J Chasovskikh S Johnson DA Johnson JA Federoff HJ Shimoji M Mhyre TR Maguire-Zeiss KA Microglial activation and antioxidant responses induced by the Parkinson’s disease protein alpha-synuclein J Neuroimmune Pharmacol 2013 8 94 117 10.1007/s11481-012-9401-0 23054368 PMC3582877 · DOI
  24. [26]Sondag CM Dhawan G Combs CK Beta amyloid oligomers and fibrils stimulate differential activation of primary microglia J Neuroinflammation 2009 6 1 10.1186/1742-2094-6-1 19123954 PMC2632990 · DOI · Paper page
  25. [27]Block ML Zecca L Hong JS Microglia-mediated neurotoxicity: uncovering the molecular mechanisms Nat Rev Neurosci 2007 8 57 69 10.1038/nrn2038 17180163 · DOI · Paper page
  26. [28]Gabellec MM Griffais R Fillion G Haour F Expression of interleukin 1 alpha, interleukin 1 beta and interleukin 1 receptor antagonist mRNA in mouse brain: regulation by bacterial lipopolysaccharide (LPS) treatment Brain Res Mol Brain Res 1995 31 122 130 10.1016/0169-328X(95)00042-Q 7476020 · DOI
  27. [29]Turrin NP Gayle D Ilyin SE Flynn MC Langhans W Schwartz GJ Plata-Salaman CR Pro-inflammatory and anti-inflammatory cytokine mRNA induction in the periphery and brain following intraperitoneal administration of bacterial lipopolysaccharide Brain Res Bull 2001 54 443 453 10.1016/S0361-9230(01)00445-2 11306198 · DOI
  28. [30]Quan N He L Lai W Endothelial activation is an intermediate step for peripheral lipopolysaccharide induced activation of paraventricular nucleus Brain Res Bull 2003 59 447 452 10.1016/S0361-9230(02)00951-6 12576141 · DOI
  29. [31]Singh AK Jiang Y How does peripheral lipopolysaccharide induce gene expression in the brain of rats? Toxicology 2004 201 197 207 10.1016/j.tox.2004.04.015 15297033 · DOI
  30. [32]Cunningham C Wilcockson DC Campion S Lunnon K Perry VH Central and systemic endotoxin challenges exacerbate the local inflammatory response and increase neuronal death during chronic neurodegeneration J Neurosci 2005 25 9275 9284 10.1523/JNEUROSCI.2614-05.2005 16207887 PMC6725757 · DOI · Paper page
  31. [33]Saper CB The dance of the perivascular and endothelial cells: mechanisms of brain response to immune signaling Neuron 2010 65 4 6 10.1016/j.neuron.2009.12.029 20152108 · DOI
  32. [34]Raivich G Bohatschek M Werner A Jones LL Galiano M Kloss CU Zhu XZ Pfeffer K Liu ZQ Lymphocyte infiltration in the injured brain: role of proinflammatory cytokines J Neurosci Res 2003 72 726 733 10.1002/jnr.10621 12774313 · DOI
  33. [37]Gao HM Jiang J Wilson B Zhang W Hong JS Liu B Microglial activation-mediated delayed and progressive degeneration of rat nigral dopaminergic neurons: relevance to Parkinson’s disease J Neurochem 2002 81 1285 1297 10.1046/j.1471-4159.2002.00928.x 12068076 · DOI
  34. [40]Lee KH Yun SJ Nam KN Gho YS Lee EH Activation of microglial cells by ceruloplasmin Brain Res 2007 1171 1 8 10.1016/j.brainres.2007.07.053 17727827 · DOI
  35. [41]Consonni A Morara S Codazzi F Grohovaz F Zacchetti D Inhibition of lipopolysaccharide-induced microglia activation by calcitonin gene related peptide and adrenomedullin Mol Cell Neurosci 2011 48 151 160 10.1016/j.mcn.2011.07.006 21803157 PMC3174421 · DOI
  36. [42]McCarthy KD de Vellis J Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue J Cell Biol 1980 85 890 902 10.1083/jcb.85.3.890 6248568 PMC2111442 · DOI
  37. [43]Kettenmann H Hanisch UK Noda M Verkhratsky A Physiology of microglia Physiol Rev 2011 91 461 553 10.1152/physrev.00011.2010 21527731 · DOI · Paper page
  38. [44]Macco R Pelizzoni I Consonni A Vitali I Giacalone G Martinelli Boneschi F Codazzi F Grohovaz F Zacchetti D Astrocytes acquire resistance to iron-dependent oxidative stress upon proinflammatory activation J Neuroinflammation 2013 10 130 10.1186/1742-2094-10-130 24160637 PMC3874684 · DOI
  39. [45]Green LC Wagner DA Glogowski J Skipper PL Wishnok JS Tannenbaum SR Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids Anal Biochem 1982 126 131 138 10.1016/0003-2697(82)90118-X 7181105 · DOI
  40. [46]Sheng W Zong Y Mohammad A Ajit D Cui J Han D Hamilton JL Simonyi A Sun AY Gu Z Hong JS Weisman GA Sun GY Pro-inflammatory cytokines and lipopolysaccharide induce changes in cell morphology, and upregulation of ERK1/2, iNOS and sPLA(2)-IIA expression in astrocytes and microglia J Neuroinflammation 2011 8 121 10.1186/1742-2094-8-121 21943492 PMC3206447 · DOI
  41. [47]Deshmukh VK Raman PH Dhuley JN Naik SR Role of ceruloplasmin in inflammation: increased serum ceruloplasmin levels during inflammatory conditions and its possible relationship with anti-inflammatory agents Pharmacol Res Commun 1985 17 633 642 10.1016/0031-6989(85)90070-0 4048245 · DOI
  42. [48]Engler R Acute-phase proteins in inflammation C R Seances Soc Biol Fil 1995 189 563 578 8564570
  43. [49]Kang JH Kim KS Choi SY Kwon HY Won MH Oxidative modification of human ceruloplasmin by peroxyl radicals Biochim Biophys Acta 2001 1568 30 36 10.1016/S0304-4165(01)00198-2 11731082 · DOI
  44. [50]Filipov NM Seegal RF Lawrence DA Manganese potentiates in vitro production of proinflammatory cytokines and nitric oxide by microglia through a nuclear factor kappa B-dependent mechanism Toxicol Sci 2005 84 139 148 10.1093/toxsci/kfi055 15601679 · DOI
  45. [51]Zhang P Lokuta KM Turner DE Liu B Synergistic dopaminergic neurotoxicity of manganese and lipopolysaccharide: differential involvement of microglia and astroglia J Neurochem 2010 112 434 443 10.1111/j.1471-4159.2009.06477.x 19895668 PMC2809141 · DOI
  46. [52]Kauppinen TM Higashi Y Suh SW Escartin C Nagasawa K Swanson RA Zinc triggers microglial activation J Neurosci 2008 28 5827 5835 10.1523/JNEUROSCI.1236-08.2008 18509044 PMC2680357 · DOI · Paper page
  47. [53]Mou YH Yang JY Cui N Wang JM Hou Y Song S Wu CF Effects of cobalt chloride on nitric oxide and cytokines/chemokines production in microglia Int Immunopharmacol 2012 13 120 125 10.1016/j.intimp.2012.03.017 22472292 · DOI
  48. [54]Zhang Y Brovkovych V Brovkovych S Tan F Lee BS Sharma T Skidgel RA Dynamic receptor-dependent activation of inducible nitric-oxide synthase by ERK-mediated phosphorylation of Ser745 J Biol Chem 2007 282 32453 32461 10.1074/jbc.M706242200 17804409 · DOI
  49. [55]Huang SH Jong AY Cellular mechanisms of microbial proteins contributing to invasion of the blood-brain barrier Cell Microbiol 2001 3 277 287 10.1046/j.1462-5822.2001.00116.x 11298651 · DOI
  50. [56]Kim KS Mechanisms of microbial traversal of the blood-brain barrier Nat Rev Microbiol 2008 6 625 634 10.1038/nrmicro1952 18604221 PMC5206914 · DOI
  51. [57]Pacher P Beckman JS Liaudet L Nitric oxide and peroxynitrite in health and disease Physiol Rev 2007 87 315 424 10.1152/physrev.00029.2006 17237348 PMC2248324 · DOI · Paper page
  52. [58]Szabo C Multiple pathways of peroxynitrite cytotoxicity Toxicol Lett 2003 140–141 105 112 10.1016/S0378-4274(02)00507-6 12676456 · DOI
  53. [59]Loeffler DA DeMaggio AJ Juneau PL Brickman CM Mashour GA Finkelman JH Pomara N LeWitt PA Ceruloplasmin is increased in cerebrospinal fluid in Alzheimer’s disease but not Parkinson’s disease Alzheimer Dis Assoc Disord 1994 8 190 197 10.1097/00002093-199408030-00005 7986488 · DOI
  54. [60]Kuhlow CJ Krady JK Basu A Levison SW Astrocytic ceruloplasmin expression, which is induced by IL-1beta and by traumatic brain injury, increases in the absence of the IL-1 type 1 receptor Glia 2003 44 76 84 10.1002/glia.10273 12951659 · DOI
  55. [61]Loeffler DA LeWitt PA Juneau PL Sima AA Nguyen HU DeMaggio AJ Brickman CM Brewer GJ Dick RD Troyer MD Kanaley L Increased regional brain concentrations of ceruloplasmin in neurodegenerative disorders Brain Res 1996 738 265 274 10.1016/S0006-8993(96)00782-2 8955522 · DOI
  56. [62]Hwang IK Yoon DK Yoo KY Eum WS Bahn JH Kim DW Kang JH Kwon HY Kang TC Choi SY Won MH Ischemia-related change of ceruloplasmin immunoreactivity in neurons and astrocytes in the gerbil hippocampus and dentate gyrus Neurochem Int 2004 44 601 607 10.1016/j.neuint.2003.10.002 15016475 · DOI
  57. [63]Marques L Auriac A Willemetz A Banha J Silva B Canonne-Hergaux F Costa L Immune cells and hepatocytes express glycosylphosphatidylinositol-anchored ceruloplasmin at their cell surface Blood Cells Mol Dis 2012 48 110 120 10.1016/j.bcmd.2011.11.005 22178061 · DOI
  58. [64]Banha J Marques L Oliveira R Martins Mde F Paixao E Pereira D Malho R Penque D Costa L Ceruloplasmin expression by human peripheral blood lymphocytes: a new link between immunity and iron metabolism Free Radic Biol Med 2008 44 483 492 10.1016/j.freeradbiomed.2007.10.032 17991445 · DOI
  59. [65]Mazumder B Mukhopadhyay CK Prok A Cathcart MK Fox PL Induction of ceruloplasmin synthesis by IFN-gamma in human monocytic cells J Immunol 1997 159 1938 1944 9257859
  60. [66]Skjorringe T Moller LB Moos T Impairment of interrelated iron- and copper homeostatic mechanisms in brain contributes to the pathogenesis of neurodegenerative disorders Front Pharmacol 2012 3 169 10.3389/fphar.2012.00169 23055972 PMC3456798 · DOI

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Authors
Lazzaro, M., Bettegazzi, B., Barbariga, M., Codazzi, F., Zacchetti, D., & Alessio, M.
Affiliations
IRCCS Ospedale San Raffaele (IT)
Corresponding author
Alessio, Massimo
ORCID
Venue
Journal of Neuroinflammation
Publisher
Springer Science and Business Media LLC
Published
2014-09-16
Volume
11
Issue
1
Pages
164-164
ISSN
1742-2094 (ISSN-L)
DOI
10.1186/s12974-014-0164-9
PMC
PMC4174266
OpenAlex
W2086219357
Cited by
33
References
7
Citations by year
2026: 1 · 2025: 5 · 2024: 1 · 2023: 4 · 2022: 6 · 2021: 3 · 2020: 3 · 2019: 1 · 2018: 2 · 2017: 4 · 2016: 1 · 2015: 2
FWCI
1.22
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Neuroinflammation (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Archivio Istituzionale della Ricerca - Università Vita-Salute San Raffaele (submittedVersion)
Funders
Michael J. Fox Foundation for Parkinson's Research (130-012/2310)
Topics
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Inflammasome and immune disorders
  • Life Sciences › Neuroscience › Neurology › Neurological Disease Mechanisms and Treatments
Keywords
ceruloplasmin, microglial activation, nitric oxide synthase, inducible nitric oxide synthase, nitric oxide production, lipopolysaccharide, interleukin-6, MIP-1α, neuroinflammation, Parkinson's disease, Alzheimer's disease, ferroxidase, neurodegenerative diseases
MeSH terms
Animals, Ceruloplasmin, Enzyme-Linked Immunosorbent Assay, Humans, Inflammation, Blotting, Western, Cytokines, Rats, Sprague-Dawley, Microglia, Reverse Transcriptase Polymerase Chain Reaction, Rats, Nitric Oxide Synthase Type II, Real-Time Polymerase Chain Reaction

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