Effects of haloperidol,clozapine and olanzapine on the survival of human neuronal and immune cells in vitro

Abstract

Cytotoxic effects on neuronal as well as on immune cells have been reported for both typical and atypical antipsychotic drugs. We evaluated the effects of different concentrations of a typical (haloperidol) and two atypical (clozapine, olanzapine) antipsychotics on the survival of human neuronal (SH-SY5Y cells) and immune cells (U937 cells) by determining the metabolic activity after 24 h of incubation by the modified tetrazolium method. The dopaminergic neuroblastoma SH-SY5Y and the lymphoma U-937 cell line are well established models for in vitro investigations. To further elucidate possible mechanisms of action we also determined the ATP content in the cultured cells.

After experimental treatment, significant effects were detected by Kruskal Wallis test for all treatment conditions. Post-hoc tests (Dunn's method) showed that haloperidol and clozapine at the two highest concentrations (25 and 50 µg/ml) caused a significant decrease of metabolic activity in both cell systems, which was also detectable after treatment with clozapine at a concentration of 12.5 µg/ml in U937 cells. In contrast, olanzapine induced a significant increase in metabolic activity of SH-SY5Y cells at all concentrations except for the concentration of 3.1 µg/ml, whereas the metabolic activity in U937 cells was increased at concentrations of 1.6 and 6.25 µg/ml. For the determination of ATP content, the LD₅₀ values of the metabolic activity were used, except for olanzapine for which no distinct LD₅₀ value was available. Significant changes were detected for all treatments and post-hoc tests revealed that haloperidol caused a significant decrease compared to the control condition in both cell systems. These findings suggest that antipsychotic substances of different classes exert differential metabolic effects in both neuronal and immune cell systems.

Keywords

antipsychotic toxicity metabolic activity ATP cell

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References

Ardizzone T.D. , Bradley R.J. , Freeman A.M. 3rd , Dwyer, DS (2001) Inhibition of glucose transport in PC12 cells by the atypical antipsychotic drugs risperidone and clozapine, and structural analogs of clozapine. Brain Res 923: 82—90

Behl C. ( 1999) Vitamin E and other antioxidants in neuroprotection. Int J Vitam Nutr Res 69: 213—219

Behl C. , Rupprecht R. , Skutella T. , Holsboer F. (1995) Haloperidol-induced cell death—mechanism and protection with vitamin E in vitro. Neuroreport 7: 360—364

Carpenter W.T. Jr , Buchanan R.W. (1994) Schizophrenia. N Engl J Med 330: 681—690

Chiarugi A. (2005) “Simple but not simpler”: toward a unified picture of energy requirements in cell death. FASEB J 19: 1783—1788

Deicken R.F. , Calabrese G. , Merrin E.L. , Fein G. , Weiner M.W. (1995) Basal ganglia phosphorous metabolism in chronic schizophrenia. Am J Psychiatry 152: 126—129

Dwyer D.S. , Pinkofsky H.B. , Liu Y. , Bradley R.J. (1999) Antipsychotic drugs affect glucose uptake and the expression of glucose transporters in PC12 cells. Prog Neuropsychopharmacol Biol Psychiatry 23: 69—80

Dwyer D.S. , Lu XH , Bradley R.J. (2003) Cytotoxicity of conventional and atypical antipsychotic drugs in relation to glucose metabolism. Brain Res 971: 31—39

Fujimoto T. , Nakano T. , Takano T. , Hokazono Y. , Asakura T. , Tsuji T. (1992) Study of chronic schizophrenics using 31P magnetic resonance chemical shift imaging. Acta Psychiatr Scand 86: 455—462.

10.

Fukuzako H. (2001) Neurochemical investigation of the schizophrenic brain by in vivo phosphorus magnetic resonance spectroscopy. World J Biol Psychiatry 2: 70—82

11.

Galili-Mosberg R. , Gil-Ad I. , Weizman A. , Melamed E. , Offen D. (2000) Haloperidol-induced neurotoxicity—possible implications for tardive dyskinesia. J Neural Transm 107: 479—490

12.

Gardner I. , Zahid N. , MacCrimmon D. , Uetrecht JP (1998) A comparison of the oxidation of clozapine and olanzapine to reactive metabolites and the toxicity of these metabolites to human leukocytes. Mol Pharmacol 53: 991—998

13.

Harris P. , Ralph P. (1985) Human leukemic models of myelomonocytic development: a review of the HL-60 and U937 cell lines. J Leukoc Biol 37: 407—422

14.

Heiser P. , Singh S. , Krieg J.C. , Vedder H. (2006) Effects of different antipsychotics and the antidepressant mirtazapine on glucose transporter mRNA levels in human blood cells. J Psychiatr Res 40: 374—379

15.

Iqbal M.M. , Rahman A. , Husain Z. , Mahmud S.Z. , Ryan W.G. , Feldman J.M. (2003) Clozapine: a clinical review of adverse effects and management. Ann Clin Psychiatry 15: 33—48

16.

Klöcking R. , Schacke M. , Wutzler P. (1995) Primärscreening antiherpetischer Verbindungen mit EZ4U. Chemotherapie J 4: 141—147

17.

Koizumi S. , Ikeda M. , Nakazawa K. , Inoue K. , Ito K. , Inoue K. (1995) Inhibition by haloperidol of adenosine 5'-triphosphate-evoked responses in rat pheochromocytoma cells. Biochem Biophys Res Commun 210: 624—630

18.

Kornhuber J. , Schultz A. , Wiltfang J. , Meineke I. , Gleiter C.H. , Zochling R. , Boissl K.W. , Leblhuber F. , Riederer P. (1999) Persistence of haloperidol in human brain tissue . Am J Psychiatry 156: 885—890

19.

Kowalski J. , Blada P. , Kucia K. , Madej A. , Herman ZS (2001) Neuroleptics normalize increased release of interleukin- 1 beta and tumor necrosis factor-alpha from monocytes in schizophrenia. Schizophr Res 50: 169—175

20.

Leykin I. , Mayer R. , Shinitzky M. (1997) Short and long-term immunosuppressive effects of clozapine and haloperidol. Immunopharmacology 37: 75—86

21.

Llorca P.M. , Chereau I. , Bayle F.J. , Lancon C. (2002) Tardive dyskinesias and antipsychotics: a review . Eur Psychiatry 17: 129—138

22.

Maurer I. , Möller H.J. (1997) Inhibition of complex I by neuroleptics in normal human brain cortex parallels the extrapyramidal toxicity of neuroleptics. Mol Cell Biochem 174: 255—259

23.

Miller D.D. (2000) Review and management of clozapine side effects . J Clin Psychiatry 61(Suppl 8): 14—17

24.

Pahlman S. , Mamaeva S. , Meyerson G. , Mattsson ME , Bjelfman C. , Ortoft E. , Hammerling U. (1990) Human neuroblastoma cells in culture: a model for neuronal cell differentiation and function. Acta Physiol Scand 592(Suppl): 25—37

25.

Pettegrew J.W. , Keshavan MS , Panchalingam K. , Strychor S. , Kaplan D.B. , Tretta M.G. , Allen M. (1991) Alterations in brain high-energy phosphate and membrane phospholipid metabolism in first-episode, drug-naive schizophrenics. A pilot study of the dorsal prefrontal cortex by in vivo phosphorus 31 nuclear magnetic resonance spectroscopy. Arch Gen Psychiatry 48: 563—568

26.

Pochet S. , Garcia-Marcos M. , Fernandez M. , Marino A. , Dehaye J.P. (2003) Regulation by clozapine of calcium handling by rat submandibular acinar cells. Cell Calcium 34: 465—475

27.

Silver M.A. , Yang Z.W. , Ganguli R. , Nimgaonkar VL (1994) An inhibitory effect of psychoactive drugs on a human neuroblastoma cell line. Biol Psychiatry 35: 824—826

28.

Skinner TE , O'Donnell J.M. , Robitaille P.M. , Nasrallah H.A. (1995) 31P MRS study of clozapine vs. haloperidol effects on brain metabolism (abstract). Biol Psychiatry 37: 648

29.

Skulachev V.P. (2006) Bioenergetic aspects of apoptosis, necrosis and mitoptosis. Apoptosis 11: 473—485

30.

Suda T. , Suzuki Y. , Matsui T. , Inoue T. , Niide O. , Yoshimaru T. , Suzuki H. , Ra C. , Ochiai T. (2005) Dapsone suppresses human neutrophil superoxide production and elastase release in a calcium-dependent manner. Br J Dermatol 152: 887—895

31.

Sunderland T. , Cohen BM (1987) Blood to brain distribution of neuroleptics. Psychiatry Res 20: 299—305

32.

Tschen A.C. , Rieder M.J. , Oyewumi L.K. , Freeman D.J. (1999) The cytotoxicity of clozapine metabolites: implications for predicting clozapine-induced agranulocytosis. Clin Pharmacol Ther 65: 526—532

33.

Tsuneizumi T. , Babb S.M. , Cohen B.M. (1992) Drug distribution between blood and brain as a determinant of antipsychotic drug effects. Biol Psychiatry 32: 817—824

34.

Vairetti M. , Feletti F. , Battaglia A. , Pamparana F. , Canonico PL , Richelmi P. , Berte F. (1999) Haloperidol-induced changes in glutathione and energy metabolism: effect of nicergoline. Eur J Pharmacol 367: 67—72

35.

Vilner B.J. , Bowen W.D. (1993) Sigma receptor-active neuroleptics are cytotoxic to C6 glioma cells in culture. Eur J Pharmacol 244: 199—201

36.

Volz H.P. , Rzanny R. , Rossger G. , Hubner G. , Kreitschmann-Andermahr I. , Kaiser W.A. , Sauer H. (1997) Decreased energy demanding processes in the frontal lobes of schizophrenics due to neuroleptics? A 31P-magneto-resonance spectroscopic study. Psychiatry Res 76: 123—129

37.

Volz H.R. , Riehemann S. , Maurer I. , Smesny S. , Sommer M. , Rzanny R. , Holstein W. , Czekalla J. , Sauer H. (2000) Reduced phosphodiesters and high-energy phosphates in the frontal lobe of schizophrenic patients: a (31)P chemical shift spectroscopic-imaging study. Biol Psychiatry 47: 954—961

38.

Yang S.B. , Proks P. , Ashcroft F.M. , Rupnik M. (2004) Inhibition of ATPsensitive potassium channels by haloperidol. Br J Pharmacol 143: 960—967