Psychiatry Research 198 (2012) 521–526
`
`Contents lists available at SciVerse ScienceDirect
`
`Psychiatry Research
`
`j o ur n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / p s y c h r e s
`
`Effects of lithium and valproate on oxidative stress and behavioral changes induced
`by administration of m-AMPH
`Dayane D. da-Rosa a, Samira S. Valvassori a, Amanda V. Steckert a,b, Felipe Ornell a, Camila L. Ferreira a,
`Jéssica Lopes-Borges a, Roger B. Varela a, Felipe Dal-Pizzol b, Monica L. Andersen c, João Quevedo a,⁎
`a Laboratory of Neurosciences and National Institute for Translational Medicine (INCT-TM), Postgraduate Program in Health Sciences, Health Sciences Unit,
`University of Southern Santa Catarina, 88806-000 Criciúma, SC, Brazil
`b Laboratory of Experimental Pathophysiology and National Institute for Translational Medicine (INCT-TM), Postgraduate Program in Health Sciences,
`Health Sciences Unit, University of Southern Santa Catarina, 88806-000 Criciúma, SC, Brazil
`c Department of Psychobiology, Universidade Federal de São Paulo, 04024-002 São Paulo, SP, Brazil
`
`a r t i c l e
`
`i n f o
`
`a b s t r a c t
`
`Article history:
`Received 4 October 2011
`Received in revised form 12 January 2012
`Accepted 18 January 2012
`
`Keywords:
`Animal model of mania
`Lithium
`Valproate
`Methamphetamine
`Oxidative stress
`
`In the last years our research group has studied and validated the animal model of mania induced by dextro-
`amphetamine (D-AMPH). Considering the lack of animal models of mania reported in the literature; this
`study evaluated the possibilities to validate the animal model induced by methamphetamine (m-AMPH).
`Then, we evaluated the effects of lithium (Li), valproate (VPA) on the behavior and parameters of oxidative dam-
`age in rat brain after administration of m-AMPH. In the prevention treatment, Wistar rats were pretreated with
`Li, VPA or saline (Sal) for 14 days, and then, between days 8 and 14, rats were treated with m-AMPH (1, 0.5 or
`0.25 mg/kg) or Sal. In the reversal treatment, rats were first given m-AMPH (0.25 mg/kg) or Sal. Locomotor be-
`havior was assessed using the open-field task and parameters of oxidative damage were measured in brain struc-
`tures. Our results show that the hyperactivity was prevented and reverted by Li and VPA only when m-AMPH
`was administered in the dose of 0.25 mg/kg. In addition, the m-AMPH in all doses administrated induced oxida-
`tive damage in both structures tested in two models. Li and VPA reversed and prevented this impairment, how-
`ever in a way dependent of cerebral area, the dose of m-AMPH and technique.
`© 2012 Elsevier Ireland Ltd. All rights reserved.
`
`1. Introduction
`
`Bipolar disorder (BD) is one of the most severe psychiatric disor-
`ders, which is associated with morbidity and mortality and psychiat-
`ric comorbidity (Kupfer, 2005; McIntyre et al., 2007). However,
`despite the severity of the disorder, its pathophysiology remains
`largely unknown. Animal models are an important tool in gaining in-
`sights into the neural mechanisms underlying BD and in assessing po-
`tential therapeutic actions of new compounds in preclinical settings
`(Lipska and Weinberger, 2000; Boksa, 2007). Animal models of
`human diseases should meet three sets of criteria: face, construct,
`and predictive validities (Einat et al., 2003; Machado-Vieira et al.,
`2004). Face validity is the ability of the model to mimic the symptoms
`of the determinate human disorder, while construct validity is the ca-
`pacity of the model to mimic some pathophysiological change found
`in the human disorder. Finally, predictive validity refers to the ability
`of the conventional drugs used to treat the disorder, prevent and/or
`reverse the symptoms induced in the model (Machado-Vieira et al.,
`2004).
`
`⁎ Corresponding author at: Laboratório de Neurociências, PPGCS, UNASAU, Universidade
`do Extremo Sul Catarinense, 88806-000 Criciúma, SC, Brazil. Fax: +55 48 3443 4817.
`E-mail address: joq@unesc.net (J. Quevedo).
`
`0165-1781/$ – see front matter © 2012 Elsevier Ireland Ltd. All rights reserved.
`doi:10.1016/j.psychres.2012.01.019
`
`Dopamine (DA) is an important neurotransmitter that is involved
`in cognition, mood and motor functions of the brain (Sibley and
`Monsma, 1992; Missale et al., 1998). Changes in the dopamine signal-
`ing are implicated in numerous neuropsychiatric disorders, including
`schizophrenia and bipolar disorder (Greengard, 2001). Many of the
`signs and symptoms of BD can be reproduced in humans and animal
`models with dopaminergic stimulants such as cocaine and amphet-
`amine. For example, amphetamine (AMPH) administration in rats in-
`duces hyperlocomotion, insomnia and increased sexual drive (Fiorino
`and Phillips, 1999) — symptoms also found in bipolar patients. Addi-
`tionally, in previous studies, our research group demonstrated that
`lithium (Li) and valproate (VPA) – two classic drugs for the treatment
`of BD – reversed and prevented the dextroamphetamine-induced hy-
`peractivity in rats (Frey et al., 2006a,b,c; Andreazza et al., 2008;
`Valvassori et al., 2010).
`AMPHs are psychostimulant drugs of the phenethylamine class that
`act by increasing DA efflux, inhibit the uptake of DA and inhibit mono-
`amine oxidase (Fischer and Cho, 1979). It is well known that the dopami-
`nergic system and oxidation of DA play pivotal roles in the neurotoxicity
`produced by AMPHs, which increase the formation of reactive
`oxygen species (Chipana et al., 2008). Dextroamphetamine (D-AMPH)
`is chemically related to methamphetamine (m-AMPH), nevertheless,
`m-AMPH is considered to be a more potent central psychostimulant
`
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`than D-AMPH (Mathias, 1998). However, behavioral and neurochemical
`differences between D-AMPH and m-AMPH remain poorly understood
`(Ricaurte et al., 1983; Kuczenski et al., 1995; Shoblock et al., 2003).
`Moreover, several lines of evidence show the oxidative damage to
`lipid and proteins as one of the possible mechanisms contributing to
`neuronal and glial impairment in BD (Kunz et al., 2008; Andreazza
`et al., 2010a,b; Steckert et al., 2010). Repeated injections of D-AMPH
`that induce hyperactivity in rodents is associated with increased in
`protein and lipid oxidative damage in brain (Gluck et al., 2001;
`Andreazza et al., 2008; Valvassori et al., 2011), supporting the view
`that oxidative stress is occurring during a manic-like state. In addi-
`tion, previous studies of our laboratory also demonstrated that Li
`and VPA prevented and reversed the D-AMPH-induced oxidative
`protein and lipid damage in the brain of rats (Frey et al., 2006c;
`Andreazza et al., 2008).
`Therapeutic development in bipolar is hampered by a lack of path-
`ophysiological model (Berk et al., 2007). In the last six years our re-
`search group has studied and validated the animal model of mania
`induced by D-AMPH, which has been well accepted in the literature
`(Frey et al., 2006c; Andreazza et al., 2008; Valvassori et al., 2010).
`Considering: 1) the lack of animal models of mania reported in the
`literature and 2) the lack of studies showing the difference between
`D-AMPH and m-AMPH; the present study aims to evaluate the possi-
`bilities to validate the animal model induced by m-AMPH. Then, we
`evaluated the effects of mood stabilizers on the behavior and parame-
`ters of oxidative damage in rat brain after administration of m-AMPH.
`
`2. Methods
`
`2.1. Animals
`
`The subjects were adult male Wistar rats (weighting 250–350 g) obtained from our
`breeding colony. Animals were housed five to a cage with food and water available ad libi-
`tum and were maintained on a 12-h light/dark cycle (lights on at 7:00 a.m.) at a temper-
`ature of 22±1 °C. All experimental procedures were performed in accordance with the
`approval of the local Ethics Committee in the use of animals at the Universidade do
`Extremo Sul Catarinense. All experiments were performed at the same time during the
`day to avoid circadian variations.
`
`2.2. Treatment protocols
`
`2.2.1. Prevention treatment
`In the prevention model, we simulated the maintenance phase of BD treatment.
`Animals were treated with Li (47.5 mg/kg i.p.), VPA (200 mg/kg i.p.) or saline twice a
`day for 14 days. Between the 8th and the 14th days, Li, VPA and saline-treated animals
`additionally received one daily intraperitoneal injection of either m-AMPH or saline.
`From this protocol 3 experiments were made . In experiment 1, a dose of 0.25 mg/kg
`of m-AMPH was administered and experimental groups were as follows: Sal+Sal, Li+
`Sal, VPA+Sal, Sal+m-AMPH 0.25 mg/kg, Li+m-AMPH 0.25 mg/kg, VPA+m-AMPH
`0.25 mg/kg (n=15 animals per group).
`In experiment 2, a dose of 0.5 mg/kg of m-AMPH was administered and experimental
`groups were as follows: Sal + Sal, Li + Sal, VPA + Sal, Sal + m-AMPH 0.5 mg/kg, Li +
`m-AMPH 0.5 mg/kg, VPA + m-AMPH 0.5 mg/kg (n = 15 animals per group).
`In experiment 3, a dose of 0.5 mg/kg of m-AMPH was administered and experi-
`mental groups were as follows: Sal+Sal, Li+Sal, VPA+Sal, Sal+m-AMPH 0.5 mg/kg,
`Li+m-AMPH 0.5 mg/kg, VPA+m-AMPH 0.5 mg/kg (n=15 animals per group).
`Locomotor activity was measured 2 h after the last injection. The rats were sacri-
`ficed by decapitation immediately after the open-field task and hippocampus, striatum
`and prefrontal were dissected, rapidly frozen and stored −70 °C until assayed.
`
`2.2.2. Reversal treatment
`In the reversal model, we reproduced the treatment of acute manic episode. The ani-
`mals received one daily intraperitoneal injection (i.p.) of m-AMPH 0.25 mg/kg or saline
`(Sal) for 14 days. On the 8th day of treatment, the animals in the saline and D-AMPH
`group were divided in three groups: 1) treatment with Li (47.5 mg/kg i.p.); 2) treatment
`with VPA (200 mg/kg i.p.) and 3) treatment with Sal for 7 days twice a day for all drugs.
`On the 15th day of treatment, the animals received a single injection of m-AMPH
`(0.25 mg/kg) or Sal and locomotor activity was assessed 2 h after the last injection. The ex-
`perimental groups were as follows: Sal+Sal, Sal+Li, Sal+VPA, m-AMPH 0.25 mg/kg+
`Sal, m-AMPH 0.25 mg/kg+Li, m-AMPH 0.25 mg/kg+VPA (n=12 animals per group).
`The rats were killed by decapitation immediately after the open-field task and pre-
`frontal, amygdale, hippocampus and striatum were dissected, rapidly frozen and
`stored at −70 °C until assayed.
`
`Note: The reversion protocol was made only with the dose of m-AMPH (0.25 mg/kg)
`in which Li and VPA could prevent the hyperactivity induced by m-AMPH.
`
`2.3. Locomotor activity
`
`Locomotor activity was assessed using the open-field task as previously described
`(Barros et al., 2002; Frey et al., 2006a,b,c). This task was performed in a 40 ×60 cm
`open field surrounded by 50 cm high walls, made of brown plywood, with the floor di-
`vided into 12 equal rectangles by black lines. The animals were gently placed on the
`left rear rectangle, and left free to explore the arena for 5 min. Crossings of the black
`lines (locomotor activity/horizontal activity) and rearings (exploratory activity/vertical
`activity) were counted.
`
`2.4. Measurement of oxidative damage markers
`
`Rats treated with m-AMPH or Saline as described above were sacrificed by decap-
`itation 2 h after the last injection and their brains were removed and dissected for eval-
`uation of oxidative damage levels in the prefrontal cortex, amygdala, hippocampus and
`striatum. Thiobarbituric acid reactive substances (TBARS) and protein carbonyl forma-
`tion were measured as previously described (Draper and Hadley, 1990; Levine et al.,
`1994).
`Note: For biochemical analysis n=5 animals per group, which were chosen ran-
`domly, were used.
`
`2.5. Thiobarbituric acid reactive substances (TBARS)
`
`The formation of TBARS during an acid-heating reaction was measured as an index
`of ROS production, which is widely adopted as a sensitive method for measurement of
`lipid peroxidation, as previously described (Draper and Hadley, 1990). Briefly, the sam-
`ples were mixed with 1 mL of trichloroacetic acid 10% (TCA) and 1 mL of thiobarbituric
`acid 0.67% (TBA), then heated in a boiling water bath for 15 min. TBARS were deter-
`mined by the absorbance at 535 nm. Results are expressed as MDA (malondialdehyde)
`equivalents (nmol/mg protein).
`
`2.6. Measurement of protein carbonyls
`
`The oxidative damage to proteins was assessed by the determination of carbonyl
`groups based on the reaction with dinitrophenylhidrazine (DNPH) as previously de-
`scribed (Levine et al., 1994). Briefly, proteins were precipitated by the addition of
`20% trichloroacetic acid and redissolved in DNPH and the absorbance read at 370 nm.
`
`2.7. Statistic analysis
`
`All analyses were performed with the statistical package for social sciences version
`19.0 (SPSS Inc. Chicago, IL, USA). All data are presented as mean+ S.E.M. To test differ-
`ences between groups, we used analysis of variance (ANOVA), followed by Tukey post-
`hoc tests. In all experiments, P values less than 0.05 were considered to indicate statistical
`significance.
`
`3. Results
`
`Administration of m-AMPH at 1 mg/kg increased locomotor activ-
`ity in rats, and both Li and VPA were not able to prevent the hyperac-
`tivity induced by m-AMPH (Fig. 1A). Administration of m-AMPH
`1 mg/kg significantly increased the carbonyl production (a direct
`index of cell protein peroxidation) in prefrontal, amygdala, hippo-
`campus and striatum of rats. In addition, Li and VPA were not able
`to prevent the m-AMPH-induced protein damage in the brain of rats
`(Fig. 1B). We also found that m-AMPH 1 mg/kg increased lipid perox-
`idation in all brain structure evaluated — as indicated by increased
`levels of TBARS. Pretreatment with Li increased and VPA decreased the
`lipid damage induced by m-AMPH 1 mg/kg in hippocampus. Converse-
`ly, Li prevent the m-AMPH-induced lipid damage in the striatum of rats.
`In prefrontal and amygdala the mood stabilizers were not able to protect
`against lipid damage induced by m-AMPH 1 mg/kg (Fig. 1C).
`Administration of m-AMPH at 0.5 mg/kg increased locomotor activity
`in rats, and both Li and VPA partially prevent the hyperactivity induced
`by m-AMPH (Fig. 2A). The administration of m-AMPH 0.5 mg/kg in rats
`increased the protein damage in all brain structures evaluated, and
`mood stabilizers were not able to prevent the m-AMPH-induced protein
`damage in the brain of rats (Fig. 2B). We verified also that administration
`of m-AMPH 0.5 mg/kg increased TBARS generation in all brain structures
`evaluated. The pretreatment with VPA significantly diminished AMPH-
`induced protein damage in all brain structures evaluated. Additionally,
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`Fig. 1. (A) Number of crossings (n=12 for each group). (B) Protein carbonyl content
`(n=5 for each group). (C) TBARS content (n=5 for each group) in the prevention
`model. Rats were pretreated with Li (47.5 mg/kg) or VPA (200 mg/kg) for seven days
`and then and then were treated with plus amphetamine until the 14th day of the ex-
`periment. Li = lithium, VPA = valproate. Bars represent means; error bars represent
`standard error of the means (SEM). *Different to the saline group. */# Different to the
`m-AMPH (1 mg/kg) group.
`
`Fig. 2. (A) Number of crossings (n=12 for each group). (B) Protein carbonyl content
`(n=5 for each group). (C) TBARS content (n=5 for each group) in the prevention
`model. Rats were pretreated with Li (47.5 mg/kg) or VPA (200 mg/kg) for seven days
`and then and then were treated with plus amphetamine until the 14th day of the ex-
`periment. Li = lithium, VPA = valproate. Bars represent means; error bars represent
`standard error of the means (SEM). *Different to the saline group. */# Different to the
`m-AMPH (0.5 mg/kg) group.
`
`Li partially prevents the lipid damage induced by m-AMPH 0.5 mg/kg in
`the hippocampus and striatum (Fig. 2C).
`Administration of m-AMPH at 0.25 mg/kg increased locomotor ac-
`tivity in rats and both Li and VPA prevented the m-AMPH-induced hy-
`peractivity in rats (Fig. 3A). Besides, the administration of m-AMPH
`0.25 mg/kg increased carbonyl group formation in all brain structure
`evaluated, and both Li and VPA prevented the protein damage induced
`by m-AMPH 0.25 mg/kg (Fig. 3B). The administration of m-AMPH
`0.25 mg/kg increased lipid damage in all brain structures evaluated.
`Pretreatment with Li protects the brain against lipid damage induced
`by m-AMPH 0.25 mg/kg in the prefrontal and partially protects the
`amygdala and the hippocampus. In addition, pretreatment with VPA
`
`prevents the m-AMPH-induced lipid damage in the prefrontal, the
`amygdala and the hippocampus (Fig. 3C).
`In the reversion protocol, the administration of m-AMPH 0.25 mg/kg
`increased locomotor activity in rats and Li and VPA reversed the hyper-
`activity induced by m-AMPH (Fig. 4A). Once more, the administration of
`m-AMPH 0.25 mg/kg increased the carbonyl group formation and Li re-
`versed m-AMPH-induced protein damage in all brain structure evaluat-
`ed. In addition, the treatment with VPA reversed the m-APH-induced
`protein damage (Fig. 4B) in the prefrontal and the hippocampus and
`partially reversed in the striatum. Besides, m-AMPH 0.25 mg/kg in-
`creased lipid damage in all brain structures evaluated. The treatment
`with Li diminished the lipid damage induced by m-AMPH 0.25 mg/kg
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`Fig. 3. (A) Number of crossings (n=12 for each group). (B) Protein carbonyl content
`(n=5 for each group). (C) TBARS content (n=5 for each group) in the prevention
`model. Rats were pretreated with Li (47.5 mg/kg) or VPA (200 mg/kg) for seven days
`and then and then were treated with plus amphetamine between the 8th and 14th
`days. Li = lithium, VPA = valproate. Bars represent means; error bars represent standard
`error of the means (SEM). *Different to the saline group. */# Different to the m-AMPH
`(0.25 mg/kg) group.
`
`in the prefrontal. Moreover, treatment with VPA diminished the
`m-AMPH-induced lipid damage in the prefrontal and amygdala (Fig. 4C).
`
`4. Discussion
`
`In the present study we observed that all doses of m-AMPH (0.25,
`0.5, or 1 mg/kg) increased locomotor activity in rats. However, this
`hyperactivity was prevented and reverted by mood stabilizers – Li
`and VPA – only when m-AMPH was administered in the dose of
`0.25 mg/kg. In the high doses of m-AMPH, 1 and 0.5 mg/kg, Li and
`VPA were not able to prevent the hyperactivity induced in the rats.
`
`Fig. 4. (A) Number of crossings (n=12 for each group). (B) Protein carbonyl content
`(n=5 for each group). (C) TBARS content (n=5 for each group) in the reversal
`model. Rats were pretreated with m-amphetamine for seven days and then treated
`with amphetamine plus Li (47.5 mg/kg) or VPA (200 mg/kg) between the 8th and
`14th days. Li = lithium, VPA = valproate. Bars represent means; error bars represent
`standard error of the means (SEM). *Different to the saline group. # Different to the
`m-AMPH (0.25 mg/kg) group.
`
`According, Ellenbroek and Cools (1990) the validity of animal
`models in psychiatric disorders should demonstrate the face, con-
`struct and predictive validities. The clinical hallmark of BD is acute
`mania (Belmaker, 2004), showing symptoms such as irritable mood,
`psychomotor activation, reduced need for sleep, and excessive in-
`volvement in potentially problematic behavior (El-Mallakh et al.,
`2003). According to several guidelines or consensus statements, lith-
`ium, anticonvulsivants such as valproic acid and carbamazepine, and
`the second generation antipsychotics are recommended for the phar-
`macological treatment of BD. Here, we demonstrated that m-AMPH
`0.25 mg/kg administration increases locomotor activity of rats, and
`the usual mood stabilizers – Li and VPA – prevents and reverses this
`hyperactivity in the animals, presenting the face and predictive valid-
`ities of the model.
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`It is known that m-AMPH and D-AMPH induces hyperactivity in
`animals by elevation in extracellular DA (Martinez et al., 2003).
`There are several studies showing the importance of dopaminergic
`transmission in the behavioral response to AMPHs. It is well de-
`scribed in the literature that amphetamine dose dependently in-
`creases extracellular levels of DA in the brain (Moghaddam and
`Bunney, 1989; Maisonneuve et al., 1990). Moreover, the blockade of
`D1 receptors in the medial prefrontal cortex attenuates AMPH- and
`m-AMPH-induced locomotor activity in the rats (Hall et al., 2009).
`In addition, intracerebral administration of 6-OHDA, which induces
`depletion of DA, prevents AMPH-induced hyperactivity (Dunnett et
`al., 1984; Banks and Gratton, 1995, King and Finlay, 1995) as well as
`the development of behavioral sensitization following repeated expo-
`sure to amphetamine (Bjijou et al., 2002).
`Joyce et al. (1995) have found that higher urinary dopamine levels
`have been associated with the emergence of manic symptoms. In ad-
`dition, studies have demonstrated dopamine receptor changes in BD
`patients (Pantazopoulos et al., 2004; Vogel et al., 2004). These studies
`suggest that the dopaminergic system may play a role in the patho-
`physiology of BD. In addition, the mechanism of AMPH toxicity has
`been suggested that involves an increase in the formation of reactive
`oxygen species (Chipana et al., 2008). It has been widely demonstrat-
`ed that the generation of reactive oxygen species (ROS) plays a criti-
`cal role in the pathophysiology of BD (Kunz et al., 2008; Andreazza et
`al., 2010a,b; Steckert et al., 2010). In the present study we also found
`that m-AMPH in all doses administrated increased protein and lipid
`damage in the brain of rats. Finally, the increase in extracellular dopa-
`mine together with the oxidative damage induced by m-AMPH, both
`present in BD, characterizes the construct validity of the model.
`Another important result found here is that Li and VPA are not able
`to prevent protein damage induced by m-AMPH 1 and 0.5 mg/kg.
`However, when given m-AMPH at 0.25 mg/kg these mood stabilizers
`prevented and reversed the m-AMPH-induced protein damage. Fur-
`thermore, Li increased the lipid damage induced by m-AMPH 1 mg/kg
`in the prefrontal and VPA partially protects lipid damage only in the
`hippocampus and striatum. When m-AMPH was administered at
`0.5 mg/kg VPA diminished the m-AMPH-induced lipid damage in all
`brain structures evaluated. However, Li diminished the lipid damage in-
`duced by m-AMPH 0.5 mg/kg only in the hippocampus and striatum.
`Additionally, Li and VPA partially protect against lipid damage induced
`by m-AMPH 0.25 mg/kg in the prefrontal, amygdala and hippocampus,
`but not in the striatum. In the reversal protocol, the lipid damage in-
`duced by m-AMPH 0.25 mg/kg was diminished by VPA in the prefrontal
`and amygdala and by Li in the prefrontal. This discrepancy between the
`prevention and reversion protocols may be due to the fact that the time
`of treatment with mood stabilizers in the prevention protocol is greater.
`In previous studies of our laboratory we demonstrated that D-AMPH
`increased TBARS levels in the prefrontal cortex, and this effect was re-
`versed by both mood stabilizers — Li and VPA. It was also observed
`that the administration of Li and VPA increased TBARS formation in
`the hippocampus of rats pretreated with D-AMPH. In the same study,
`it has been demonstrated that the VPA pretreatment protects the pre-
`frontal and the hippocampus against D-AMPH-induced lipid damage.
`However, Li partially prevented D-AMPH-induced lipid peroxidation in
`the rat hippocampus but augmented in the prefrontal (Frey et al.,
`2006c). Together, the previous studies and the present study demon-
`strate that the effect of mood stabilizers depends on the experimental
`protocol and the brain area analyzed. Regions of the central nervous
`systems differentially respond to insults (Sullivan et al., 2004) and
`TBARS formation was evaluated from different brain regions, that in
`part represent different cell types. Furthermore, within a homogeneous
`population of cells, there is heterogeneity in terms of physiological and
`metabolic characteristics (Lai et al., 1977; Sims, 1991; Sonnewald et al.,
`1998).
`It is noteworthy that in the animal model of mania induced by
`D-AMPH protein damage was not observed (Frey et al., 2006c).
`
`Here we show that m-AMPH 0.25 mg/kg induces protein damage,
`which is reversed and prevented by administration of mood stabilizers.
`In a postmortem study, it was observed that protein oxidation and nitra-
`tion increased in the prefrontal cortex of bipolar patients (Andreazza et
`al., 2010a,b). In addition, Andreazza et al. (2009) found a significant in-
`crease in 3-nitrotyrosine (marker of protein damage) levels in peripher-
`al blood of patients in the early and late stages of bipolar disorder. From
`these evidences, we suggest that the animal model of mania induced by
`m-AMPH is more effective than D-AMPH in mimicking the oxidative
`brain damage seen in bipolar patients.
`
`5. Conclusions
`
`In conclusion, in the present study we demonstrate that the mood
`stabilizers Li and VPA reversed and prevented m-AMPH-induced hy-
`peractivity when m-AMPH was administered at 0.25 mg/kg. In addi-
`tion, the hyperactivity induced by m-AMPH was associated with
`lipid and protein damage, which Li and VPA were able to reverse
`and prevent depending on the experimental protocol and the brain
`region evaluated. Then, we propose a new model of mania induced
`by m-AMPH, being that this model has face, predictive and construct
`validities.
`
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