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Journal of Neuroimmune Pharmacology

, Volume 7, Issue 1, pp 187–201 | Cite as

Clozapine Protects Dopaminergic Neurons from Inflammation-Induced Damage by Inhibiting Microglial Overactivation

  • Xiaoming Hu
  • Hui Zhou
  • Dan Zhang
  • Sufen Yang
  • Li Qian
  • Hung-Ming Wu
  • Po-See Chen
  • Belinda Wilson
  • Hui-Ming Gao
  • Ru-band Lu
  • Jau-Shyong HongEmail author
ORIGINAL ARTICLE

Abstract

Increasing evidence suggests a possible involvement of neuroinflammation in some psychiatric disorders, and also pharmacological reports indicate that anti-inflammatory effects are associated with therapeutic actions of psychoactive drugs, such as anti-depressants and antipsychotics. The purpose of this study was to explore whether clozapine, a widely used antipsychotic drugs, displays anti-inflammatory and neuroprotective effects. Using primary cortical and mesencephalic neuron-glia cultures, we found that clozapine was protective against inflammation-related neurodegeneration induced by lipopolysaccharide (LPS). Pretreatment of cortical or mesencephalic neuron–glia cultures with clozapine (0.1 or 1 μM) for 24 h attenuated LPS-induced neurotoxicity. Clozapine also protected neurons against 1-methyl-4-phenylpyridinium + (MPP + )-induced neurotoxicity, but only in cultures containing microglia, indicating an indispensable role of microglia in clozapine-afforded neuroprotection. Further observation revealed attenuated LPS-induced microglial activation in primary neuron-glia cultures and in HAPI microglial cell line with clozapine pretreatment. Clozapine ameliorated the production of microglia-derived superoxide and intracellular reactive oxygen species (ROS), as well as the production of nitric oxide and TNF-α following LPS. In addition, the protective effect of clozapine was not observed in neuron-glia cultures from mice lacking functional NADPH oxidase (PHOX), a key enzyme for superoxide production in immune cells. Further mechanistic studies demonstrated that clozapine pretreatment inhibited LPS-induced translocation of cytosolic subunit p47phox to the membrane in microglia, which was most likely through inhibiting the phosphoinositide 3-kinase (PI3K) pathway. Taken together, this study demonstrates that clozapine exerts neuroprotective effect via the attenuation of microglia activation through inhibition of PHOX-generated ROS production and suggests potential use of antipsychotic drugs for neuroprotection.

Keywords

Clozapine Microglia NADPH oxidase Neurodegeneration Neuroinflammation 

Notes

Acknowledgements

We would like to thank Anthony Lockhart for assistance with animal colony management and maintenance of the timed pregnant mice. This research was supported by the Intramural Research Program of the National Institute of Health, the National Institute of Environmental Health Sciences.

Conflict of interest

The authors declare that they have nocompeting interests or conflicts of interest.

References

  1. Adams RA, Bauer J, Flick MJ, Sikorski SL, Nuriel T, Lassmann H, Degen JL, Akassoglou K (2007) The fibrin-derived gamma377-395 peptide inhibits microglia activation and suppresses relapsing paralysis in central nervous system autoimmune disease. J Exp Med 204:571–582PubMedCrossRefGoogle Scholar
  2. Altshuler LL, Casanova MF, Goldberg TE, Kleinman JE (1990) The hippocampus and parahippocampus in schizophrenia, suicide, and control brains. Arch Gen Psychiatry 47:1029–1034PubMedCrossRefGoogle Scholar
  3. Anderson KE, Boyle KB, Davidson K, Chessa TA, Kulkarni S, Jarvis GE, Sindrilaru A, Scharffetter-Kochanek K, Rausch O, Stephens LR, Hawkins PT (2008) CD18-dependent activation of the neutrophil NADPH oxidase during phagocytosis of Escherichia coli or Staphylococcus aureus is regulated by class III but not class I or II PI3Ks. Blood 112:5202–5211PubMedCrossRefGoogle Scholar
  4. Araki N, Johnson MT, Swanson JA (1996) A role for phosphoinositide 3-kinase in the completion of macropinocytosis and phagocytosis by macrophages. J Cell Biol 135:1249–1260PubMedCrossRefGoogle Scholar
  5. Babior BM (1999) NADPH oxidase: an update. Blood 93:1464–1476PubMedGoogle Scholar
  6. Bai O, Wei Z, Lu W, Bowen R, Keegan D, Li XM (2002) Protective effects of atypical antipsychotic drugs on PC12 cells after serum withdrawal. J Neurosci Res 69:278–283PubMedCrossRefGoogle Scholar
  7. Baune BT, Eyre H (2010) Anti-inflammatory effects of antidepressant and atypical antipsychotic medication for the treatment of major depression and comorbid arthritis: a case report. J Med Case Reports 4:6PubMedCrossRefGoogle Scholar
  8. Block ML, Hong JS (2007) Chronic microglial activation and progressive dopaminergic neurotoxicity. Biochem Soc Trans 35:1127–1132PubMedCrossRefGoogle Scholar
  9. Block ML, Zecca L, Hong JS (2007) Microglia-mediated neurotoxicity: uncovering the molecular mechanisms. Nat Rev Neurosci 8:57–69PubMedCrossRefGoogle Scholar
  10. Brustolim D, Ribeiro-dos-Santos R, Kast RE, Altschuler EL, Soares MB (2006) A new chapter opens in anti-inflammatory treatments: the antidepressant bupropion lowers production of tumor necrosis factor-alpha and interferon-gamma in mice. Int Immunopharmacol 6:903–907PubMedCrossRefGoogle Scholar
  11. Byne W, Buchsbaum MS, Mattiace LA, Hazlett EA, Kemether E, Elhakem SL, Purohit DP, Haroutunian V, Jones L (2002) Postmortem assessment of thalamic nuclear volumes in subjects with schizophrenia. Am J Psychiatr 159:59–65PubMedCrossRefGoogle Scholar
  12. Dobos N, Korf J, Luiten PG, Eisel UL (2010) Neuroinflammation in Alzheimer’s disease and major depression. Biol Psychiatr 67:503–504CrossRefGoogle Scholar
  13. Doorduin J, de Vries EF, Willemsen AT, de Groot JC, Dierckx RA, Klein HC (2009) Neuroinflammation in schizophrenia-related psychosis: a PET study. J Nucl Med 50:1801–1807PubMedCrossRefGoogle Scholar
  14. Ellson CD, Gobert-Gosse S, Anderson KE, Davidson K, Erdjument-Bromage H, Tempst P, Thuring JW, Cooper MA, Lim ZY, Holmes AB, Gaffney PR, Coadwell J, Chilvers ER, Hawkins PT, Stephens LR (2001) PtdIns(3)P regulates the neutrophil oxidase complex by binding to the PX domain of p40(phox). Nat Cell Biol 3:679–682PubMedCrossRefGoogle Scholar
  15. Gao HM, Hong JS (2008) Why neurodegenerative diseases are progressive: uncontrolled inflammation drives disease progression. Trends Immunol 29:357–365PubMedCrossRefGoogle Scholar
  16. Gao HM, Jiang J, Wilson B, Zhang W, Hong JS, Liu B (2002) Microglial activation-mediated delayed and progressive degeneration of rat nigral dopaminergic neurons: relevance to Parkinson’s disease. J Neurochem 81:1285–1297PubMedCrossRefGoogle Scholar
  17. Gao HM, Liu B, Zhang W, Hong JS (2003) Critical role of microglial NADPH oxidase-derived free radicals in the in vitro MPTP model of Parkinson’s disease. FASEB J 17:1954–1956PubMedGoogle Scholar
  18. Gao X, Hu X, Qian L, Yang S, Zhang W, Zhang D, Wu X, Fraser A, Wilson B, Flood PM, Block M, Hong JS (2008) Formyl-methionyl-leucyl-phenylalanine-induced dopaminergic neurotoxicity via microglial activation: a mediator between peripheral infection and neurodegeneration? Environ Health Perspect 116:593–598PubMedCrossRefGoogle Scholar
  19. Gorlach A, Kietzmann T, Hess J (2002) Redox signaling through NADPH oxidases: involvement in vascular proliferation and coagulation. Ann N Y Acad Sci 973:505–507PubMedCrossRefGoogle Scholar
  20. Gross A, Joffe G, Joutsiniemi SL, Nyberg P, Rimon R, Appelberg B (2003) Decreased production of reactive oxygen species by blood monocytes caused by clozapine correlates with EEG slowing in schizophrenic patients. Neuropsychobiology 47:73–77PubMedCrossRefGoogle Scholar
  21. Herken H, Uz E, Ozyurt H, Sogut S, Virit O, Akyol O (2001) Evidence that the activities of erythrocyte free radical scavenging enzymes and the products of lipid peroxidation are increased in different forms of schizophrenia. Mol Psychiatr 6:66–73CrossRefGoogle Scholar
  22. Hulshoff Pol HE, Schnack HG, Bertens MG, van Haren NE, van der Tweel I, Staal WG, Baare WF, Kahn RS (2002) Volume changes in gray matter in patients with schizophrenia. Am J Psychiatr 159:244–250PubMedCrossRefGoogle Scholar
  23. Kang UG, Seo MS, Roh MS, Kim Y, Yoon SC, Kim YS (2004) The effects of clozapine on the GSK-3-mediated signaling pathway. FEBS Lett 560:115–119PubMedCrossRefGoogle Scholar
  24. Kapur S, Zipursky RB, Remington G (1999) Clinical and theoretical implications of 5-HT2 and D2 receptor occupancy of clozapine, risperidone, and olanzapine in schizophrenia. Am J Psychiatr 156:286–293PubMedGoogle Scholar
  25. Kreutzberg GW (1996) Microglia: a sensor for pathological events in the CNS. Trends Neurosci 19:312–318PubMedCrossRefGoogle Scholar
  26. Kropp S, Kern V, Lange K, Degner D, Hajak G, Kornhuber J, Ruther E, Emrich HM, Schneider U, Bleich S (2005) Oxidative stress during treatment with first- and second-generation antipsychotics. J Neuropsychiatry Clin Neurosci 17:227–231PubMedCrossRefGoogle Scholar
  27. Liu B, Hong JS (2003) Role of microglia in inflammation-mediated neurodegenerative diseases: mechanisms and strategies for therapeutic intervention. J Pharmacol Exp Ther 304:1–7PubMedCrossRefGoogle Scholar
  28. Lu XH, Dwyer DS (2005) Second-generation antipsychotic drugs, olanzapine, quetiapine, and clozapine enhance neurite outgrowth in PC12 cells via PI3K/AKT, ERK, and pertussis toxin-sensitive pathways. J Mol Neurosci 27:43–64PubMedCrossRefGoogle Scholar
  29. Mahadik SP, Mukherjee S, Scheffer R, Correnti EE, Mahadik JS (1998) Elevated plasma lipid peroxides at the onset of nonaffective psychosis. Biol Psychiatry 43:674–679PubMedCrossRefGoogle Scholar
  30. McGeer EG, McGeer PL (1998) The importance of inflammatory mechanisms in Alzheimer disease. Exp Gerontol 33:371–378PubMedCrossRefGoogle Scholar
  31. Minghetti L (2005) Role of inflammation in neurodegenerative diseases. Curr Opin Neurol 18:315–321PubMedCrossRefGoogle Scholar
  32. Moller T (2002) Calcium signaling in microglial cells. Glia 40:184–194PubMedCrossRefGoogle Scholar
  33. Munn NA (2000) Microglia dysfunction in schizophrenia: an integrative theory. Med Hypotheses 54:198–202PubMedCrossRefGoogle Scholar
  34. Nair TR, Christensen JD, Kingsbury SJ, Kumar NG, Terry WM, Garver DL (1997) Progression of cerebroventricular enlargement and the subtyping of schizophrenia. Psychiatr Res 74:141–150CrossRefGoogle Scholar
  35. Paterson GJ, Ohashi Y, Reynolds GP, Pratt JA, Morris BJ (2006) Selective increases in the cytokine, TNFalpha, in the prefrontal cortex of PCP-treated rats and human schizophrenic subjects: influence of antipsychotic drugs. J Psychopharmacol 20:636–642PubMedCrossRefGoogle Scholar
  36. Qian L, Flood PM, Hong JS (2010) Neuroinflammation is a key player in Parkinson’s disease and a prime target for therapy. J Neural Transm 117:971–979PubMedCrossRefGoogle Scholar
  37. Qin L, Liu Y, Cooper C, Liu B, Wilson B, Hong JS (2002) Microglia enhance beta-amyloid peptide-induced toxicity in cortical and mesencephalic neurons by producing reactive oxygen species. J Neurochem 83:973–983PubMedCrossRefGoogle Scholar
  38. Qing H, Xu H, Wei Z, Gibson K, Li XM (2003) The ability of atypical antipsychotic drugs vs. haloperidol to protect PC12 cells against MPP + −induced apoptosis. Eur J Neurosci 17:1563–1570PubMedCrossRefGoogle Scholar
  39. Radewicz K, Garey LJ, Gentleman SM, Reynolds R (2000) Increase in HLA-DR immunoreactive microglia in frontal and temporal cortex of chronic schizophrenics. J Neuropathol Exp Neurol 59:137–150PubMedGoogle Scholar
  40. Reddy RD, Yao JK (1996) Free radical pathology in schizophrenia: a review. Prostaglandins Leukot Essent Fatty Acids 55:33–43PubMedCrossRefGoogle Scholar
  41. Seidel A, Arolt V, Hunstiger M, Rink L, Behnisch A, Kirchner H (1996) Major depressive disorder is associated with elevated monocyte counts. Acta Psychiatr Scand 94:198–204PubMedCrossRefGoogle Scholar
  42. Shao Z, Dyck LE, Wang H, Li XM (2006) Antipsychotic drugs cause glial cell line-derived neurotrophic factor secretion from C6 glioma cells. J Psychiatr Neurosci 31:32–37Google Scholar
  43. Shin SY, Choi BH, Ko J, Kim SH, Kim YS, Lee YH (2006) Clozapine, a neuroleptic agent, inhibits Akt by counteracting Ca2+/calmodulin in PTEN-negative U-87MG human glioblastoma cells. Cell Signal 18:1876–1886PubMedCrossRefGoogle Scholar
  44. Song C, Lin A, Kenis G, Bosmans E, Maes M (2000) Immunosuppressive effects of clozapine and haloperidol: enhanced production of the interleukin-1 receptor antagonist. Schizophr Res 42:157–164PubMedCrossRefGoogle Scholar
  45. Tian W, Li XJ, Stull ND, Ming W, Suh CI, Bissonnette SA, Yaffe MB, Grinstein S, Atkinson SJ, Dinauer MC (2008) Fc gamma R-stimulated activation of the NADPH oxidase: phosphoinositide-binding protein p40phox regulates NADPH oxidase activity after enzyme assembly on the phagosome. Blood 112:3867–3877PubMedCrossRefGoogle Scholar
  46. van Berckel BN, Bossong MG, Boellaard R, Kloet R, Schuitemaker A, Caspers E, Luurtsema G, Windhorst AD, Cahn W, Lammertsma AA, Kahn RS (2008) Microglia activation in recent-onset schizophrenia: a quantitative (R)-[11 C]PK11195 positron emission tomography study. Biol Psychiatr 64:820–822CrossRefGoogle Scholar
  47. van Haren NE, Hulshoff Pol HE, Schnack HG, Cahn W, Mandl RC, Collins DL, Evans AC, Kahn RS (2007) Focal gray matter changes in schizophrenia across the course of the illness: a 5-year follow-up study. Neuropsychopharmacology 32:2057–2066PubMedCrossRefGoogle Scholar
  48. Vieira OV, Botelho RJ, Rameh L, Brachmann SM, Matsuo T, Davidson HW, Schreiber A, Backer JM, Cantley LC, Grinstein S (2001) Distinct roles of class I and class III phosphatidylinositol 3-kinases in phagosome formation and maturation. J Cell Biol 155:19–25PubMedCrossRefGoogle Scholar
  49. Wierzba-Bobrowicz T, Lewandowska E, Lechowicz W, Stepien T, Pasennik E (2005) Quantitative analysis of activated microglia, ramified and damage of processes in the frontal and temporal lobes of chronic schizophrenics. Folia Neuropathol 43:81–89PubMedGoogle Scholar
  50. Yasuda Y, Shinagawa R, Yamada M, Mori T, Tateishi N, Fujita S (2007) Long-lasting reactive changes observed in microglia in the striatal and substantia nigral of mice after 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Brain Res 1138:196–202PubMedCrossRefGoogle Scholar
  51. Yirmiya R, Pollak Y, Barak O, Avitsur R, Ovadia H, Bette M, Weihe E, Weidenfeld J (2001) Effects of antidepressant drugs on the behavioral and physiological responses to lipopolysaccharide (LPS) in rodents. Neuropsychopharmacology 24:531–544PubMedCrossRefGoogle Scholar
  52. Zarifkar A, Choopani S, Ghasemi R, Naghdi N, Maghsoudi AH, Maghsoudi N, Rastegar K, Moosavi M (2010) Agmatine prevents LPS-induced spatial memory impairment and hippocampal apoptosis. Eur J Pharmacol 634:84–88PubMedCrossRefGoogle Scholar
  53. Zhang XY, Zhou DF, Cao LY, Zhang PY, Wu GY (2003) Elevated blood superoxide dismutase in neuroleptic-free schizophrenia: association with positive symptoms. Psychiatr Res 117:85–88CrossRefGoogle Scholar
  54. Zhang W, Shin EJ, Wang T, Lee PH, Pang H, Wie MB, Kim WK, Kim SJ, Huang WH, Wang Y, Zhang W, Hong JS, Kim HC (2006) 3-Hydroxymorphinan, a metabolite of dextromethorphan, protects nigrostriatal pathway against MPTP-elicited damage both in vivo and in vitro. FASEB J 20:2496–2511PubMedCrossRefGoogle Scholar
  55. Zhang D, Hu X, Wei SJ, Liu J, Gao H, Qian L, Wilson B, Liu G, Hong JS (2008) Squamosamide derivative FLZ protects dopaminergic neurons against inflammation-mediated neurodegeneration through the inhibition of NADPH oxidase activity. J Neuroinflammation 5:21PubMedCrossRefGoogle Scholar
  56. Zhang D, Hu X, Qian L, Chen SH, Zhou H, Wilson B, Miller DS, Hong JS (2011) Microglial MAC1 receptor and PI3K are essential in mediating beta-amyloid peptide-induced microglial activation and subsequent neurotoxicity. J Neuroinflammation 8:3PubMedCrossRefGoogle Scholar
  57. Zhuang X, Oosting RS, Jones SR, Gainetdinov RR, Miller GW, Caron MG, Hen R (2001) Hyperactivity and impaired response habituation in hyperdopaminergic mice. Proc Natl Acad Sci USA 98:1982–1987PubMedCrossRefGoogle Scholar
  58. Zipursky RB, Lim KO, Sullivan EV, Brown BW, Pfefferbaum A (1992) Widespread cerebral gray matter volume deficits in schizophrenia. Arch Gen Psychiatr 49:195–205PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC (outside the USA)  2011

Authors and Affiliations

  • Xiaoming Hu
    • 1
    • 3
  • Hui Zhou
    • 1
  • Dan Zhang
    • 1
  • Sufen Yang
    • 1
  • Li Qian
    • 1
  • Hung-Ming Wu
    • 1
    • 2
  • Po-See Chen
    • 1
  • Belinda Wilson
    • 1
  • Hui-Ming Gao
    • 1
  • Ru-band Lu
    • 2
  • Jau-Shyong Hong
    • 1
    Email author
  1. 1.Neuropharmacology Section, Laboratory of Toxicology and PharmacologyNational Institute of Environmental Health Sciences, NIHResearch Triangle ParkUSA
  2. 2.Institute of Behavioral Medicine and Department of Psychiatry, College of Medicine & HospitalNational Cheng-Kung UniversityTainanTaiwan
  3. 3.Department of Neurology and Pittsburgh Institute of Neurodegenerative DiseasesUniversity of Pittsburgh School of MedicinePittsburghUSA

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