Hostname: page-component-745bb68f8f-grxwn Total loading time: 0 Render date: 2025-02-06T08:11:56.315Z Has data issue: false hasContentIssue false
  • English
  • Français

In vitro effect of antipsychotics on brain energy metabolism parameters in the brain of rats

Published online by Cambridge University Press:  22 February 2013

Giselli Scaini ,
Natália Rochi ,
Meline O. S. Morais ,
Débora D. Maggi ,
Bruna T. De-Nês ,
João Quevedo  and
Emilio L. Streck
Giselli Scaini
Affiliation:
Laboratório de Bioenergética, Programa de Pós-graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma, SC, Brazil Instituto Nacional de Ciência e Tecnologia Translacional em Medicina (INCT-TM), Porto Alegre, RS, Brazil
Natália Rochi
Affiliation:
Laboratório de Bioenergética, Programa de Pós-graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma, SC, Brazil Instituto Nacional de Ciência e Tecnologia Translacional em Medicina (INCT-TM), Porto Alegre, RS, Brazil
Meline O. S. Morais
Affiliation:
Laboratório de Bioenergética, Programa de Pós-graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma, SC, Brazil Instituto Nacional de Ciência e Tecnologia Translacional em Medicina (INCT-TM), Porto Alegre, RS, Brazil
Débora D. Maggi
Affiliation:
Laboratório de Bioenergética, Programa de Pós-graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma, SC, Brazil Instituto Nacional de Ciência e Tecnologia Translacional em Medicina (INCT-TM), Porto Alegre, RS, Brazil
Bruna T. De-Nês
Affiliation:
Laboratório de Bioenergética, Programa de Pós-graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma, SC, Brazil Instituto Nacional de Ciência e Tecnologia Translacional em Medicina (INCT-TM), Porto Alegre, RS, Brazil
João Quevedo
Affiliation:
Instituto Nacional de Ciência e Tecnologia Translacional em Medicina (INCT-TM), Porto Alegre, RS, Brazil Laboratório de Neurociências, Programa de Pós-graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma, SC, Brazil
Emilio L. Streck*
Affiliation:
Laboratório de Bioenergética, Programa de Pós-graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma, SC, Brazil Instituto Nacional de Ciência e Tecnologia Translacional em Medicina (INCT-TM), Porto Alegre, RS, Brazil
*
Emilio L. Streck, Laboratório de Bioenergética, Universidade do Extremo Sul Catarinense, 88806-000, Criciúma, SC, Brazil. Tel: +554834312539; E-mail: emiliostreck@gmail.com
Rights & Permissions [Opens in a new window]

Abstract

Objective

Typical and atypical antipsychotic drugs have been shown to have different clinical, biochemical and behavioural profiles. It is well described that impairment of metabolism, especially in the mitochondria, leads to oxidative stress and neuronal death and has been implicated in the pathogenesis of a number of diseases in the brain. In this context, we investigated the in vitro effect of antipsychotic drugs on energy metabolism parameters in the brain of rats.

Methods

Clozapine (0.1, 0.5 and 1.0 mg/ml), olanzapine (0.1, 0.5 and 1.0 mg/ml) and aripiprazole (0.05, 0.15 and 0.3 mg/ml) were suspended in buffer and added to the reaction medium containing rat tissue homogenates and the respiratory chain complexes, succinate dehydrogenase and creatine kinase (CK) activities were evaluated.

Results

Our results showed that olanzapine and aripriprazole increased the activities of respiratory chain complexes. On the other hand, complex IV activity was inhibited by clozapine, olanzapine and aripriprazole. CK activity was increased by clozapine at 0.5 and 1.0 mg/ml in prefrontal cortex, cerebellum, striatum, hippocampus and posterior cortex of rats. Moreover, olanzapine and aripiprazole did not affect CK activity.

Conclusion

In this context, if the hypothesis that metabolism impairment is involved in the pathophysiology of neuropsychiatric disorders is correct and these results also occur in vivo, we suggest that olanzapine may reverse a possible diminution of metabolism.

Keywords

aripiprazoleclozapinecreatine kinasemitochondrial respiratory chainolanzapine
Type
Original Articles
Information
Acta Neuropsychiatrica , Volume 25 , Issue 1 , February 2013 , pp. 18 - 26
Copyright
Scandinavian College of Neuropsychopharmacology 2013

Significant outcomes

  • • Olanzapine and aripriprazole increased the activities of respiratory chain complexes I, II and II–III.

  • • Complex IV activity was inhibited by clozapine, olanzapine and aripriprazole.

  • • Creatine kinase (CK) activity was increased by clozapine in prefrontal cortex, cerebellum, striatum, hippocampus and posterior cortex of rats.

  • • Olanzapine and aripiprazole did not affect CK activity.

Limitations

  • • It is necessary to investigate other important steps of energy metabolism.

Introduction

Neuropsychiatric disorders such as schizophrenia, depression and bipolar disorder have been related to dysfunction in the brain metabolism Reference Albert, Hemmings and Adamo1, Reference Konradi, Eaton, MacDonald, Walsh, Benes and Heckers2, Reference Prince, Yassin and Oreland3, and on neurotrophic factor expression Reference Angelucci, Mathe and Aloe4, Reference Angelucci, Oliviero and Pilato5, Reference Duman6, Reference Duman7, alterations in the neuronal function and survival Reference Schlattner and Wallimann8, Reference Velligan, Newcomer and Pultz9, as well as abnormal synaptogenesis and neurotransmission Reference Albert, Hemmings and Adamo1, Reference Duman6, Reference Duman7. The metabolism dysfunction includes mitochondrial impairment Reference Konradi, Eaton, MacDonald, Walsh, Benes and Heckers2, Reference Kato and Kato10, increase in reactive oxygen species (ROS) production and expression of biochemical markers of cellular degeneration Reference Schlattner and Wallimann8, Reference Fatemi, Laurence and Araghi-Niknam11, Reference Machado-Vieira, Lara and Portela12, Reference Rothermundt, Missler and Arolt13, Reference Schroeter, Abdul-Khaliq and Fruhauf14.

Atypical antipsychotics such as clozapine and olanzapine exhibit serotonin/dopamine antagonistic properties associated with fewer extrapyramidal symptoms than conventional antipsychotics Reference Velligan, Newcomer and Pultz9, Reference Carlson, Cavazzoni, Berg, We, Beasley and Kane15. Aripiprazole is an atypical neuroleptic that exhibits a high binding affinity for D2 and D3 receptors, a moderate affinity for D4 receptors and a low affinity for D1 receptors. Preclinical studies have also indicated that aripiprazole has a relatively high affinity for serotonin 5-HT2A and 5-HT1A receptors. It displays partial agonist activity at the 5-HT1A receptor and antagonistic activity at the 5-HT2A receptor Reference Burris, Molski and Xu16. In humans, recent evidence suggests that schizophrenic patients treated with atypical antipsychotics may present a better performance in cognitive tasks when compared to patients treated with typical antipsychotics Reference Velligan, Newcomer and Pultz9, Reference Jordan, Koprivica, Chen, Tottori, Kikuchi and Altar17.

Mitochondria are intracellular organelles which play a crucial role in adenosine triphosphate (ATP) production Reference Beuzen, Taylor, Wesnes and Wood18. Mitochondrial oxidative phosphorylation is the major ATP-producing pathway Reference Calabrese, Scapagnini, Giuffrida-Stella, Bates and Clark19. Energy, in the form of ATP, is obtained in the mitochondria through a series of reactions in which electrons liberated from reducing substrates NADH and FADH2 are delivered to O2 via the electron transport chain, which consists of four multimeric complexes (I, II, III and IV) plus two small electron carriers, coenzyme Q (or ubiquinone) and cytochrome c. The energy obtained by the reactions of the electron transport chain is used to pump protons from the mitochondrial matrix into the intermembrane space located between the inner and outer mitochondrial membranes. This process creates an electrochemical proton gradient, which is utilised by complex V (or ATP synthase) to catalyse the formation of ATP by the adenosine diphosphate (ADP) Reference Madrigal, Olivenza and Moro20, Reference Fattal, Budur, Vaughan and Franco21. Another way to produce ATP is through CK (EC 2.7.3.2) which is a crucial enzyme for high energy-consuming tissues like brain, skeletal muscle and heart. This enzyme works as a buffering system of cellular ATP levels, playing a central role in energy metabolism. CK catalyses the reversible transfer of the phosphoryl group from phosphocreatine to ADP, regenerating ATP Reference Wallimann, Wyss, Brdiczka, Nicolay and Eppenberger22, Reference Bessman and Carpenter23. In addition, CK may be considered as a marker of creatine/phosphocreatine system functioning in brain Reference Tsuji, Nakamura, Ogata, Shibata and Kataoka24. In this context, it has been widely shown diminution of CK activity may potentially impair energy homeostasis, contributing to cell death Reference Tomimoto, Yamamoto, Homburger and Yanagihara25, Reference Aksenov, Aksenova, Butterfield and Markesbery26, Reference David, Shoemaker and Haley27, Reference Gross, Bak and Ingwall28, Reference Streck, Amboni and Scaini29.

The brain contains a large number of mitochondria, being more susceptible to fluctuations on aerobic metabolism. Neurons have a high metabolic rate, limited stores of glucose or glycogen, low anaerobe capacity and a near exclusive dependence on glucose as an energy substrate Reference Lee, Grabb, Zipfel and Choi30. It is well described that impairment of energy production caused by mitochondrial dysfunction has been implicated in the pathogenesis of a number of diseases, including neurological conditions such as dementia, cerebral ischaemia, Alzheimer's disease and Parkinson's disease Reference Machado-Vieira, Lara and Portela12, Reference Streck, Amboni and Scaini29, Reference Lee, Grabb, Zipfel and Choi30, Reference Blass31, Reference Brennan, Bird and Aprille32, Reference Gruno, Peet and Tein33, Reference Schurr34. Neuropsychiatry disorders such as schizophrenia, depression and bipolar disorder have also been related to dysfunction on neurotrophic factor expression Reference Angelucci, Mathe and Aloe4, Reference Angelucci, Oliviero and Pilato5, Reference Duman6, Reference Duman7, dysfunction in the brain metabolism Reference Albert, Hemmings and Adamo1, Reference Prabakaran, Swatton and Ryan35 alterations in the neuronal function and survival Reference Rothermundt, Ponath and Arolt36, Reference Weis and Llenos37 as well as abnormal synaptogenesis and neurotransmission Reference Albert, Hemmings and Adamo1, Reference Duman6, Reference Duman7. In addition, Assis and colleagues Reference Assis, Scaini and Di-Pietro38 have recently shown that brain CK activity is altered by antipsychotics. Moreover, succinate dehydrogenase activity was inhibited in cerebellum and striatum after olanzapine and clozapine administration, respectively, whereas aripiprazole increased the enzyme in prefrontal cortex Reference Streck, Rezin, Barbosa, Assis, Grandi and Quevedo39. Some studies also reported that chronic exposure to clozapine resulted in significant changes in the activities of antioxidant enzymes and oxidative damage in rat brain, but no change was observed with olanzapine administration Reference Polydoro, Schröder and Lima40.

Considering that reduction of brain energy metabolism is related to neurological dysfunction and that some neuroleptics alter oxidative stress parameters and cognitive function, in this study we evaluated the effects in vitro of some antipsychotics, named clozapine, olanzapine and aripiprazole on mitochondrial respiratory chain and CK activities in brain (hippocampus, striatum, cerebellum, posterior cortex and prefrontal cortex) of rats.

Materials and methods

Animals

Adult male Wistar rats (250–300 g) were obtained from the Central Animal House of University of Extremo Sul Catarinense, Criciúma, SC, Brazil. They were caged in groups of five with free access to food and water and were maintained on a 12-h light:dark cycle (lights on 07:00 h), at a temperature of 22 ± 1°C. All experimental procedures were carried out in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and the Brazilian Society for Neuroscience and Behavior recommendations for animal care, with the approval of UNESC Ethics Committee.

Drugs

Clozapine was purchased from Novartis Biociências AS, São Paulo, Brazil (Leponex®). Olanzapine was provided from Eli Lilly do Brasil Ltda, São Paulo, Brazil (Zyprexa®). Aripiprazole was purchased from Bristol-Myers Squibb, São Paulo, Brazil (Abilify®).

Tissue and homogenate preparation

On the day of the experiments the animals were killed by decapitation (n = 6), the brain was removed and cerebellum, hippocampus, striatum, prefrontal cortex and posterior cortex were homogenised (1:10, w/v) in SETH buffer, pH 7.4 (250 mM sucrose, 2 mM ethylenediaminetetraacetic acid, 10 mM Trizma base, 50 IU/ml heparin). The homogenates were centrifuged at 800 × g for 10 min and the supernatants kept at −70 °C until used for enzymes activity determination. The maximal period between homogenate preparation and enzyme analysis was always less than 5 days. Protein content was determined by the method described by Lowry and colleagues Reference Lowry, Rosebrough, Farr and Randall41 using bovine serum albumin as standard. Clozapine (0.1, 0.5 and 1.0 mg/ml), olanzapine (0.1, 0.5 and 1.0 mg/ml) and aripiprazole (0.05, 0.15 and 0.3 mg/ml) were dissolved on the day of the experiments in the incubation medium (buffer) used for each technique.

Activities of mitochondrial respiratory chain enzymes

In the day of the assays, the samples were frozen and thawed in hypotonic assay buffer three times to fully expose the enzymes to substrates and achieve maximal activities. NADH dehydrogenase (complex I) was evaluated according to the method described by Cassina and Radi Reference Cassina and Radi42 by the rate of NADH-dependent ferricyanide reduction at 420 nm. The activities of succinate: 2,6-dichlorophenolindophenol (DCIP) oxidoreductase (complex II) and succinate: cytochrome c oxidoreductase (complexes II–III) were determined according to the method of Fischer and colleagues Reference Fischer, Ruitenbeek and Berden43. Complex II activity was measured by following the decrease in absorbance due to the reduction of 2,6-DCIP at 600 nm. The activity of complexes II–III was measured by cytochrome c reduction from succinate. The activity of cytochrome c oxidase (complex IV) was assayed according to the method described by Rustin and colleagues Reference Rustin, Chretien and Bourgeron44, measured by following the decrease in absorbance due to the oxidation of previously reduced cytochrome c at 550 nm. The activities of the mitochondrial respiratory chain complexes were expressed as nmol/min/mg protein.

CK activity

CK activity was measured in brain homogenates pre-treated with 0.625 mM lauryl maltoside. Briefly, the reaction mixture consisted of 60 mM Tris–HCl, pH 7.5, containing 7.0 mM phosphocreatine, 9.0 mM MgSO4 and approximately 0.4–1.2 mg protein in a final volume of 100 ml. After 15 min of preincubation at 37°C, the reaction was then started by the addition of 4.0 mM of ADP and stopped after 10 min by the addition of 20 ml of 50 mM phydroxy-mercuribenzoic acid. The creatine formed was estimated according to the colorimetric method of Hughes Reference Hughes45. The colour was developed by the addition of 100 ml 2% α-naphtol and 100 ml 0.05% diacetyl in a final volume of 1.0 ml and read spectrophotometrically after 20 min at λ = 540 nm. Results were expressed as units/min × mg protein.

Statistical analysis

Data were analysed by one-way analysis of variance followed by the Tukey test when F was significant and are expressed as mean ± SD. All analyses were performed using the Statistical Package for the Social Science (SPSS) software (IBM Corporation, NY, USA).

Results

In this study, we evaluated the effects of clozapine, olanzapine and aripiprazole on activities of mitochondrial respiratory chain enzymes and CK activity in some brain areas (hippocampus, striatum, cerebellum, prefrontal cortex and posterior cortex) of rats. It can be seen in Fig. 1 that complex I activity was increased in prefrontal cortex and striatum by olanzapine at 0.5 mg/ml and higher concentrations (prefrontal cortex p = 0.005 and p = 0.001; striatum p = 0.004 and p = 0.000). As shown in Fig. 1, olanzapine at 0.5 and 1.0 mg/ml increased complex II activity in cerebellum, striatum and posterior cortex (cerebellum p = 0.003 and p = 0.000; striatum p = 0.002 and p = 0.000; posterior cortex p = 0.002 and p = 0.000), whereas prefrontal cortex and hippocampus were not affected. Furthermore, olanzapine at 0.5 mg/ml and higher concentrations increased the activity of complexes II–III in prefrontal cortex, striatum and posterior cortex (prefrontal cortex p = 0.012 and p = 0.002; striatum p = 0.000 and p = 0.000; posterior cortex p = 0.000 and p = 0.000). In cerebellum and hippocampus, olanzapine increased the activity of complexes II–III only in the higher dose (cerebellum p = 0.000; hippocampus p = 0.000) (Fig.1). On the other hand, olanzapine inhibited complex IV activity in cerebellum only in the higher dose (cerebellum p = 0.001), without affecting prefrontal cortex, striatum, hippocampus and posterior cortex (Fig.1).

Figure 1 In vitro effect of olanzapine on the activities of complexes I (a), II (b), II–III (c) and IV (d) in the brain of rats. Data were analysed by Tukey test and are expressed as mean ± SD (n = 6). Different from control, *p < 0.05.

The next set of experiments was performed in order to evaluate the in vitro effect of clozapine on the respiratory chain complex activities. Fig. 2 shows that clozapine did not affect the activities of complexes I, II and II–III in the cerebral regions evaluated in this study (Fig. 2, respectively). In contrast, clozapine at 0.1 mg/ml and higher concentrations inhibited complex IV activity in prefrontal cortex and hippocampus (prefrontal cortex p = 0.002, p = 0.000 and p = 0.004; hippocampus p = 0.000, p = 0.001 and p = 0.000), on the other hand, as clozapine at 0.5 and 1.0 mg/ml decreased complex IV activity in cerebellum (p = 0.001 and p = 0.006), but striatum and posterior cortex were not affected (Fig. 2).

Figure 2 In vitro effect of clozapine on the activities of complexes I (a), II (b), II–III (c) and IV (d) in the brain of rats. Data were analysed by Tukey test and are expressed as mean ± SD (n = 6). Different from control, *p < 0.05.

Fig. 3 shows that aripiprazole 0.15 and 0.3 mg/ml increased complex I activity in striatum (p = 0.006 and p = 0.003), but not affect prefrontal cortex, cerebellum, hippocampus and posterior cortex (Fig. 3). On the other hand, aripriprazole at 0.15 and 0.3 mg/ml increased complex II activity in posterior cortex (p = 0.002 and p = 0.007). In cerebellum aripiprazole increased complex II activity only in the higher dose (p = 0.020). Moreover, prefrontal cortex, striatum and hippocampus were not affected (Fig. 3). In contrast, the activity of complexes II–III was increased by aripiprazole (0.3 mg/ml) in striatum and hippocampus (striatum p = 0.038; hippocampus p = 0.004) (Fig. 3). On the other hand, aripiprazole inhibited complex IV activity in prefrontal cortex only in the higher dose (prefrontal cortex p = 0.014), without affecting cerebellum, striatum, hippocampus and posterior cortex (Fig. 3).

Figure 3 In vitro effect of aripriprazole on the activities of complexes I (a), II (b), II–III (c) and IV (d) in the brain of rats. Data were analysed by Tukey test and are expressed as mean ± SD (n = 6). Different from control, *p < 0.05.

Finally, we tested the influence of olanzapine, clozapine and aripiprazole on the CK activity. It can be seen in Fig. 4 that CK activity was increased by clozapine at 0.5 and 1.0 mg/ml in prefrontal cortex, cerebellum, striatum, hippocampus and posterior cortex of rats (prefrontal cortex p = 0.000 and p = 0.000; cerebellum p = 0.015 and p = 0.000; striatum p = 0.018 and p = 0.000; hippocampus p = 0.002 and p = 0.009; posterior cortex p = 0.001 and p = 0.000). Olanzapine and aripiprazole did not affect CK activity in either tested structure (Fig. 4, respectively).

Figure 4 In vitro effect of olanzapine (a), clozapine (b) and aripiprazole (c) on CK activity in the brain of rats. Data were analysed by Tukey test and are expressed as mean ± SD (n = 6). Different from control, *p < 0.05.

Discussion

Dysfunction in brain metabolism is related to neuropsychiatric disorders, such as schizophrenia, depression and bipolar disorder Reference Albert, Hemmings and Adamo1, Reference Konradi, Eaton, MacDonald, Walsh, Benes and Heckers2, Reference Prince, Yassin and Oreland3, Reference Kato and Kato10. Some evidence also point to the possibility that drugs used in the treatment of such disorders modulate energy metabolism, especially by increasing it. In this context, we reported recently that mood stabilisers prevent and reverse inhibitory effect on cytrate synthase activity caused by amphetamine in an animal model of mania Reference Corrêa, Amboni and Assis46. Regarding antipsychotics, reports from literature show that the atypical antipsychotic olanzapine only inhibited succinate dehydrogenase (SDH) in cerebellum and aripiprazole increased the same enzyme in prefrontal cortex, whereas clozapine inhibited the enzyme only in striatum.

Volz and colleagues Reference Volz, Riehemann and Maurer47 observed decreased levels of ATP in the frontal lobes of antipsychotic-free patients. Furthermore, reports from literature show that ATP levels may inversely correlate with the degree of negative symptoms in schizophrenic patients Reference Volz, Rzanny and Rossger48. Additionally, two groups reported a lower ATP concentration in the basal ganglia of medicated schizophrenic patients, suggesting an imbalance between ATP production and ATP utilisation by oxidative phosphorylation in this disorder Reference Deicken, Calabrese, Merrin, Fein and Weiner49. Andreassen and colleagues Reference Andreassen, Ferrante, Beal and Jorgensen50 verified that mild mitochondrial impairment in combination with neuroleptics results in striatal excitotoxic neurodegeneration, which may underlie the development of persistent vacuous chewing movements in rats.

It is well described that some side-effects of antipsychotics limited their long-term use and are probably associated to oxidative stress Reference Cadet and Lohr51, Reference Lohr, Cadet and Lohr52 and metabolism impairment Reference Arnaiz, Coronel and Boveris53. The biochemical and physiological characteristics of the brain, with high-unsaturated phospholipids content and energy requirement, make this organ particularly susceptible to free radical-mediated damage Reference Madrigal, Olivenza and Moro20, Reference Gross, Bak and Ingwall28. It is well known that mitochondrial oxidative phosphorylation system generates ROS and the electron transport chain, mainly complexes I and III, is vulnerable to damage by them Reference Adam-Vizi54, Reference Torres, Torres and Gamaro55. In this context, several studies show that antipsychotics cause oxidative stress Reference Jordan, Koprivica, Chen, Tottori, Kikuchi and Altar17, Reference Polydoro, Schröder and Lima40. Augmented ROS production causes defects in the mitochondrial genome, leading to impaired oxidative phosphorylation, which not only limits ATP generation but also further promotes ROS production Reference Gruno, Peet and Tein33. Moreover, oxidative damage can be cause or consequence of mitochondrial dysfunction Reference Madrigal, Olivenza and Moro20, Reference Torres, Torres and Gamaro55.

In this study, we observed that olanzapine and aripriprazole increased the activities of respiratory chain complexes. On the other hand, complex IV activity was inhibited by clozapine, olanzapine and aripriprazole. Prince and colleagues Reference Prince, Yassin and Oreland3 evaluated regional alterations in neuronal functional activity in rat brain using complex IV histochemistry following chronic treatment with haloperidol, fluphenazine and clozapine for 28 days. The authors verified that increases in complex IV activity were evident in the frontal cortex of all treated animals. Moreover, clozapine and fluphenazine, but not haloperidol, caused significant increases in complex IV activity in other areas. The authors also suggest that neuroleptics achieve their therapeutic effects primarily via an enhancement of brain function in the frontal cortex, but also point to other brain regions which may be involved in the actions of these drugs Reference Prince, Yassin and Oreland3. In contrast, ours result suggests that chronic treatment using clozapine increases complex IV activity by an indirect mechanism, probably mediated by signal transduction and/or gene expression pathways.

The creatine/phosphocreatine/CK system is important for normal energy homeostasis by exerting several integrated functions, such as temporary energy buffering, metabolic capacity, energy transfer and metabolic control Reference Schlattner and Wallimann8, Reference Khuchua, Qin and Boero56. The brain of adult rats, like other tissues with high and variable rates of ATP metabolism, presents high phosphocreatine concentration and CK activity. It has been widely shown that a decrease in CK activity is associated with a neurodegenerative pathway that results in neuronal loss following brain ischaemia Reference Tomimoto, Yamamoto, Homburger and Yanagihara25, neurodegenerative diseases Reference Aksenov, Aksenova, Butterfield and Markesbery26, Reference David, Shoemaker and Haley27 and other pathological states Reference Streck, Amboni and Scaini29, Reference Lee, Grabb, Zipfel and Choi30.

Considering that CK is sensitive to free radicals Reference Wolosker, Panizzutti and Englender57 and that we have recently shown that antipsychotics lead to oxidative stress, we speculate that the enzyme inhibition in vivo may occur by the oxidation of thiol groups of its structure. In this context, Assis and colleagues Reference Assis, Scaini and Di-Pietro38 observed that chronic administration of clozapine inhibited CK activity in cerebellum and prefrontal cortex. Moreover, the increase in vitro CK activity by clozapine differs from in vivo studies; thus we suggest that the increased activity occur by direct mechanisms, whereas inhibition in vivo may occur by the oxidation of thiol groups of its structure.

Acknowledgements

This research was supported by grants from Programa de Pós-graduação em Ciências da Saúde – Universidade do Extremo Sul Catarinense (UNESC), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and Instituto Nacional de Ciência e Tecnologia Translacional em Medicina (INCT).

References

1Albert, KA, Hemmings, HC, Adamo, AIet al.Evidence for decreased DARPP-32 in the prefrontal cortex of patients with schizophrenia. Arch Gen Psychiatry 2002;59:705–712.Google Scholar
2Konradi, C, Eaton, M, MacDonald, ML, Walsh, J, Benes, FM, Heckers, S.Molecular evidence for mitochondrial dysfunction in bipolar disorder. Arch Gen Psychiatry 2004;1:300–308.CrossRefGoogle Scholar
3Prince, JA, Yassin, MS, Oreland, L.A histochemical demonstration of altered cytochrome oxidase activity in the rat brain by neuroleptics. Eur Neuropsychopharmacol 1998;8:1–6.Google Scholar
4Angelucci, F, Mathe, AA, Aloe, L.Neurotrophic factors and CNS disorders: findings in rodent models of depression and schizophrenia. Prog Brain Res 2004;146:151–165.Google Scholar
5Angelucci, F, Oliviero, A, Pilato, Fet al.Transcranial magnetic stimulation and BDNF plasma levels in amyotrophic lateral sclerosis. Neuroreport 2004;15:717–720.Google Scholar
6Duman, RS.Pathophysiology of depression: the concept of synaptic plasticity. Eur Psychiatry 2002;17:306–310.Google Scholar
7Duman, RS.Synaptic plasticity and mood disorders. Mol Psychiatry 2002;7:29–34.Google Scholar
8Schlattner, U, Wallimann, T.Octamers of mitochondrial creatine kinase isoenzymes differ in stability and membrane binding. J Biol Chem 2000;275:17314–17320.Google Scholar
9Velligan, DI, Newcomer, J, Pultz, Jet al.Does cognitive function improve with quetiapine in comparison to haloperidol? Schizophr Res 2002;53:239–248.Google Scholar
10Kato, T, Kato, N.Mitochondrial dysfunction in bipolar disorder. Bipolar Disord 2000;2:180–190.CrossRefGoogle ScholarPubMed
11Fatemi, SH, Laurence, JA, Araghi-Niknam, Met al.Glial fibrillary acidic protein is reduced in cerebellum of subjects with major depression, but not schizophrenia. Schizophr Res 2004;69:317–323.Google Scholar
12Machado-Vieira, R, Lara, DR, Portela, LVet al.Elevated serum S100B protein in drug-free bipolar patients during first manic episode: a pilot study. Eur Neuropsychopharmacol 2002;12:269–272.Google Scholar
13Rothermundt, M, Missler, U, Arolt, Vet al.Increased S100B blood levels in unmedicated and treated schizophrenic patients are correlated with negative symptomatology. Mol Psychiatry 2001;6:445–449.Google Scholar
14Schroeter, ML, Abdul-Khaliq, H, Fruhauf, Set al.Serum S100B is increased during early treatment with antipsychotics and in deficit schizophrenia. Schizophr Res 2003;62:231–236.Google Scholar
15Carlson, CD, Cavazzoni, PA, Berg, PH, We, HI, Beasley, CM, Kane, JM.An integrated analysis of acute treatment-emergent extrapyramidal syndrome in patients with schizophrenia during olanzapine clinical trials: comparisons with placebo, haloperidol, risperidone, or clozapine. J Clin Psychiatry 2003;64:898–906.Google Scholar
16Burris, KD, Molski, TF, Xu, Cet al. Aripiprazole, a novel antipsychotic, is a high-affinity partial agonist at human dopamine D2 receptors. J Pharmacol Exp Ther 2002; 302:381–389.Google Scholar
17Jordan, S, Koprivica, V, Chen, R, Tottori, K, Kikuchi, T, Altar, CA.The antipsychotic aripiprazole is a potent, partial agonist at the human 5-HT1A receptor. Eur J Pharmacol 2002;441:137–140.CrossRefGoogle ScholarPubMed
18Beuzen, JN, Taylor, N, Wesnes, K, Wood, A.A comparison of the effects of olanzapine, haloperidol and placebo on cognitive and psychomotor functions in healthy elderly volunteers. J Psychopharmacol 1999;13:152–158.Google Scholar
19Calabrese, V, Scapagnini, G, Giuffrida-Stella, AM, Bates, TE, Clark, JB.Mitochondrial involvement in brain function and dysfunction: relevance to aging, neurodegenerative disorders and longevity. Neurochem Res 2001;26: 739–764.Google Scholar
20Madrigal, JL, Olivenza, R, Moro, MAet al.Glutathione depletion, lipid peroxidation and mitochondrial dysfunction are induced by chronic stress in rat brain. Neuropsychopharmacology 2001;24:420–429.Google Scholar
21Fattal, O, Budur, K, Vaughan, AJ, Franco, K.Review of the literature on major mental disorders in adult patients with mitochondrial diseases. Psychosomatics 2006;47:1–7.Google Scholar
22Wallimann, T, Wyss, M, Brdiczka, D, Nicolay, K, Eppenberger, HM.Intracellular compartmentation, structure and function of creatine kinase isoenzymes in tissues with high and fluctuating energy demands: the ‘phosphocreatine circuit' for cellular energy homeostasis. Biochem J 1992;281:21–40.Google Scholar
23Bessman, SP, Carpenter, CL.The creatine-creatine phosphate energy shuttle. Annu Rev Biochem 1985;54:831–865.Google Scholar
24Tsuji, T, Nakamura, Y, Ogata, T, Shibata, T, Kataoka, K.Rapid decrease in ATP content without recovery phase duringglutamate-induced cell death in cultured spinal neurons. Brain Res 1994;62:289–292.Google Scholar
25Tomimoto, H, Yamamoto, K, Homburger, HA, Yanagihara, T.Immunoelectron microscopic investigation of creatine kinase BB-isoenzyme after cerebral ischemia in gerbils. Acta Neuropathol 1993;86:447–455.Google Scholar
26Aksenov, M, Aksenova, M, Butterfield, DA, Markesbery, WR.Oxidative modification of creatine kinase BB in Alzheimer's disease brain. J Neurochem 2000;74: 2520–2527.Google Scholar
27David, S, Shoemaker, M, Haley, BE.Abnormal properties of creatine kinase in alzheimer's disease brain: correlation of reduced enzyme activity and active site photolabeling with aberrant cytosol-membrane partitioning. Mol Brain Res 1998;54:276–287.CrossRefGoogle ScholarPubMed
28Gross, WL, Bak, MI, Ingwall, JSet al.Nitric oxide inhibits creatine kinase and regulates rat heart contractile reserve. Proc Natl Acad Sci U S A 1996;93:5604–5609.Google Scholar
29Streck, EL, Amboni, G, Scaini, Get al.Brain creatine kinase activity in an animal model of mania. Life Sci 2008;82:424–429.Google Scholar
30Lee, JM, Grabb, MC, Zipfel, GJ, Choi, DW.Brain tissue responses to ischemia. J Clin Invest 2000;106:723–731.Google Scholar
31Blass, JP.Brain metabolism and brain disease: is metabolic deficiency the proximate cause of Alzheimer dementia? J Neurosci Res 2001;66:851–856.CrossRefGoogle ScholarPubMed
32Brennan, WA, Bird, ED, Aprille, JR.Regional mitochondrial respiratory activity in Huntington's disease brain. J Neurochem 1985;44:1948–1950.CrossRefGoogle ScholarPubMed
33Gruno, M, Peet, N, Tein, Aet al.Atrophic gastritis: deficient complex I of the respiratory chain in the mitochondria of corpus mucosal cells. J Gastroenterol 2008;43:780–788.Google Scholar
34Schurr, A.Energy metabolism, stress hormones and neural recovery from cerebral ischemia/hypoxia. Neurochem Int 2002;41:1–8.Google Scholar
35Prabakaran, S, Swatton, JE, Ryan, MMet al.Mitochondrial dysfunction in schizophrenia: evidence for compromised brain metabolism and oxidative stress. Mol Psychiatry 2004;9:684–697.CrossRefGoogle ScholarPubMed
36Rothermundt, M, Ponath, G, Arolt, V.S100B in schizophrenic psychosis. Int Rev Neurobiol 2004;59:445–470.Google Scholar
37Weis, S, Llenos, IC.GFAP-immunopositive astrocytes in schizophrenia. Schizophr Res 2004;67:293–295.Google Scholar
38Assis, LC, Scaini, G, Di-Pietro, PBet al.Effect of antipsychotics on creatine kinase activity in rat brain. Basic Clin Pharmacol Toxicol 2007;101:315–319.Google Scholar
39Streck, EL, Rezin, GT, Barbosa, LM, Assis, LC, Grandi, E, Quevedo, J.Effect of antipsychotics on succinate dehydrogenase and cytochrome oxidase activities in rat brain. Naunyn Schmiedebergs Arch Pharmacol 2007;376: 127–133.Google Scholar
40Polydoro, M, Schröder, N, Lima, MNet al.Haloperidol- and clozapine-induced oxidative stress in the rat brain. Pharmacol Biochem Behav 2004;78:751–756.Google Scholar
41Lowry, OH, Rosebrough, NJ, Farr, AL, Randall, RJ.Protein measurement with the folin phenol reagent. J Biol Chem 1951;193:265–267.Google Scholar
42Cassina, A, Radi, R.Differential inhibitory action of nitric oxide and peroxynitrite on mitochondrial electron transport. Arch Biochem Biophys 1996;328:309–316.Google Scholar
43Fischer, JC, Ruitenbeek, W, Berden, JAet al.Differential investigation of the capacity of succinate oxidation in human skeletal muscle. Clin Chim Acta 1985;153:23–36.Google Scholar
44Rustin, P, Chretien, D, Bourgeron, Tet al.Biochemical and molecular investigations in respiratory chain deficiencies. Clin Chim Acta 1994;228:35–51.Google Scholar
45Hughes, BP.A method for estimation of serum creatine kinase and its use in comparing creatine kinase and aldolase activity in normal and pathologic sera. Clin Chim Acta 1962;7:597–604.Google Scholar
46Corrêa, C, Amboni, G, Assis, LCet al.Effects of lithium and valproate on hippocampus citrate synthase activity in an animal model of mania. Prog Neuropsychopharmacol Biol Psychiatry 2007;31:887–891.CrossRefGoogle Scholar
47Volz, HR, Riehemann, S, Maurer, Iet al.Reduced phosphodiesters and high-energy phosphates in the frontal lobe of schizophrenic patients: A (31)P chemical shift spectroscopic-imaging study. Biol Psychiatry 2000;47: 954–961.CrossRefGoogle ScholarPubMed
48Volz, HP, Rzanny, R, Rossger, Get al.Decreased energy demanding processes in the frontal lobes of schizophrenics due to neuroleptics? A 31Pmagneto-resonance spectroscopic study. Psychiatry Res 1997;76:123–129.Google Scholar
49Deicken, RF, Calabrese, G, Merrin, EL, Fein, G, Weiner, MW.Basal ganglia phosphorous metabolism in chronic schizophrenia. Am J Psychiatry 1995;152:126–129.Google Scholar
50Andreassen, OA, Ferrante, RJ, Beal, MF, Jorgensen, HA.Oral Dyskinesias and striatal lesions in rats after long-term co-treatment with haloperidol and 3-nitropropionic acid. Neuroscience 1998;87:639–648.Google Scholar
51Cadet, JL, Lohr, JB.Possible involvement of free radicals in neuroleptic-induced movement disorders. Evidence from treatment of tardive dyskinesia with vitamin E. Ann N Y Acad Sci 1989;570:176–185.CrossRefGoogle ScholarPubMed
52Lohr, JB, Cadet, JL, Lohr, MAet al.Vitamin E in the treatment of tardive dyskinesia: the possible involvement of free radical mechanisms. Schizophr Bull 1988;14:291–296.Google Scholar
53Arnaiz, SL, Coronel, MF, Boveris, A.Nitric oxide, superoxide and hydrogen peroxide production in brain mitochondria after haloperidol treatment. Nitric Oxide 1999;3: 235–243.Google Scholar
54Adam-Vizi, V.Production of reactive oxygen species in brain mitochondria: contribution by electron transport chain and non-electron transport chain sources. Antioxid Redox Signal 2005;7:1140–1149.Google Scholar
55Torres, RL, Torres, ILS, Gamaro, GDet al.Lipid peroxidation and total radical-trapping potential of the lungs of rats submitted to chronic and sub-chronic stress. Braz J Med Biol Res 2004;37:185–192.CrossRefGoogle ScholarPubMed
56Khuchua, ZA, Qin, W, Boero, Jet al.Octamer formation and coupling of cardiac sarcomeric mitochondrial creatine kinase are mediated by charged N-terminal residues. J Biol Chem 1998;273:22990–22996.Google Scholar
57Wolosker, H, Panizzutti, R, Englender, S.Inhibition of creatine kinase by S-nitrosoglutathione. FEBS Lett 1996;392:274–276.Google Scholar
Figure 0

Figure 1 In vitro effect of olanzapine on the activities of complexes I (a), II (b), II–III (c) and IV (d) in the brain of rats. Data were analysed by Tukey test and are expressed as mean ± SD (n = 6). Different from control, *p < 0.05.

Figure 1

Figure 2 In vitro effect of clozapine on the activities of complexes I (a), II (b), II–III (c) and IV (d) in the brain of rats. Data were analysed by Tukey test and are expressed as mean ± SD (n = 6). Different from control, *p < 0.05.

Figure 2

Figure 3 In vitro effect of aripriprazole on the activities of complexes I (a), II (b), II–III (c) and IV (d) in the brain of rats. Data were analysed by Tukey test and are expressed as mean ± SD (n = 6). Different from control, *p < 0.05.

Figure 3

Figure 4 In vitro effect of olanzapine (a), clozapine (b) and aripiprazole (c) on CK activity in the brain of rats. Data were analysed by Tukey test and are expressed as mean ± SD (n = 6). Different from control, *p < 0.05.