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Original Paper
Effects of cannabidiol on amphetamine-
induced oxidative stress generation in an
animal model of mania Journal of Psychopharmacology
25(2) 274–279
!The Author(s) 2011
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DOI: 10.1177/0269881109106925
jop.sagepub.com
Samira S Valvassori
1,5
, Guilherme Elias
1,5
, Bruna de Souza
1,5
,
Fabrı
´cia Petronilho
1,5
, Felipe Dal-Pizzol
1,5
, Fla
´vio Kapczinski
2,5
,
Clarissa Trzesniak
3,5
, Vitor Tumas
3,5
, Serdar Dursun
4,5
, Marcos Hortes
Nisihara Chagas
3,5
, Jaime EC Hallak
3,5
, Antonio W Zuardi
3,5
,
Joa
˜o Quevedo
1,5
and Jose
´AS Crippa
3,5
Abstract
Cannabidiol (CBD), a Cannabis sativa constituent, may present a pharmacological profile similar to mood stabilizing drugs, in addition to anti-oxidative
and neuroprotective properties. The present study aims to directly investigate the effects of CBD in an animal model of mania induced by
D-amphetamine (D-AMPH). In the first model (reversal treatment), rats received saline or D-AMPH (2 mg/kg) once daily intraperitoneal (i.p.) for
14 days, and from the 8th to the 14th day, they were treated with saline or CBD (15, 30 or 60 mg/kg) i.p. twice a day. In the second model (prevention
treatment), rats were pretreated with saline or CBD (15, 30, or 60 mg/kg) regime i.p. twice a day, and from the 8th to the 14th day, they also received
saline or D-AMPH i.p. once daily. In the hippocampus CBD (15 mg/kg) reversed the D-AMPH-induced damage and increased (30 mg/kg) brain-derived
neurotrophic factor (BDNF) expression. In the second experiment, CBD (30 or 60 mg/kg) prevented the D-AMPH-induced formation of carbonyl group in
the prefrontal cortex. In the hippocampus and striatum the D-AMPH-induced damage was prevented by CBD (15, 30 or 60 mg/kg). At both treatments
CBD did not present any effect against D-AMPH-induced hyperactivity. In conclusion, we could not observe effects on locomotion, but CBD protect
against D-AMPH-induced oxidative protein damage and increased BDNF levels in the reversal model and these effects vary depending on the brain
regions evaluated and doses of CBD administered.
Keywords
BDNF, bipolar disorder, cannabidiol, mania, oxidative stress
Introduction
Bipolar disorder (BD) is a relatively common condition
afflicting approximately 1% of the general population, and
is considered a chronic disease that may require lifetime treat-
ment. According to several guidelines or consensus state-
ments, lithium, anticonvulsivants such as valproic acid and
carbamazepine, and the second generation antipsychotics are
recommended for the pharmacological treatment of BD.
Antidepressants such as selective serotonin reuptake inhibi-
tors (SSRIs), can be added if mood stabilizers are not suffi-
cient, particularly in the depressive phase. Although there
have been substantial advances in the pharmacotherapeutics
of this condition over the last 10–15 years, the benefits have
been predominantly in terms of tolerability and safety
(Mitchell and Malhi, 2006). All of such medications have
important disadvantages such as careful dosage control, low
adherence, recurrence of symptoms on withdrawn, important
risks during pregnancy and breastfeeding and many
unwanted side-effects (Goodwin, 2003). In addition, BD
symptoms are often poorly controlled by the existing stan-
dard medications and frequently involve a combination of
drugs. Thus, the investigation of newer pharmacological
agents for use in the acute and maintenance phases of BD
is clearly necessary.
It is well known that cannabis can cause adverse effects,
including psychosis, anxiety and mania (Frankhauser, 2002;
Zuardi et al., 2006a,b), although anecdotal reports suggest
that some patients claim that the use of herbal cannabis
1
Laborato
´rio de Neurocie
ˆncias, Programa de Po
´s-Graduac¸a˜o em Cie
ˆncias da
Sau
´de, Unidade Acade
ˆmica de Cie
ˆncias da Sau
´de, Universidade do Extremo
Sul Catarinense, Criciu
´ma, SC, Brasil.
2
Bipolar Disorders Programme and Laboratory of Molecular Psychiatry,
Centro de Pesquisas, Hospital de Clı
´nicas de Porto Alegre, Porto Alegre,
RS, Brasil.
3
Department of Behavioral Neurosciences; Division of Psychiatry, Ribeira˜o
Preto Medical School, University of Sa˜o Paulo, Ribeira˜o Preto SP, Brazil.
4
Department of Psychiatry, University of Alberta, Edmonton, Alberta,
Canada.
5
INCT Translational Medicine, CNPq, Brazil.
Corresponding author:
Jose
´AS Crippa, Department of Behavioral Neurosciences; Division of
Psychiatry, Ribeira˜o Preto Medical School, University of Sa˜o Paulo,
Ribeira˜o Preto SP, Brazil
Email: jcrippa@fmrp.usp.br
at UNIV EXTREMO SUL CATARINENSE P on January 23, 2015jop.sagepub.comDownloaded from
preparations may alleviate depression and/or mania symp-
toms (Ashton et al., 2005; Ware et al., 2005). However,
there is no substantial epidemiological evidence that cannabis
abuse serves as a kind of self-medication for BD.
Cannabidiol (CBD), one of the main constituents from the
cannabis plant, was previously proposed as a cannabinoid
devoid of psychopharmacological activity. CBD can antago-
nize some behavioral effects of 9-THC, such as catalepsy and
impairment of variable-interval schedule performance
(Formukong et al., 1988; Zuardi and Karniol, 1983).
Moreover, CBD blocks psychotomimetic and anxiogenic
effects of 9-THC in humans (Karniol et al., 1974; Zuardi et
al., 1982), an effect that probably involves pharmacodynamic
rather than pharmacokinetic interactions (Hunt et al., 1981).
The antiepileptic effect of CBD was one of the first phar-
macological actions described with such cannabinoid, both in
experimental animals by a variety of procedures (Ashton and
Young, 2003; Carlini et al., 1973; Izquierdo et al., 1973; Porter
et al., 1999; Turkanis et al., 1974) and later in epilepsy patients
who do not achieve complete control of (disabling) seizures
(Cunha et al., 1980). Potential antidepressant (Musty et al.,
2002), hypnotic (Monti, 1977) and anxiolytic (Crippa et al.,
2004; Fusar-Poli et al., 2009; Guimara
˜es et al., 1990; Moreira
et al., 2006; Onaivi et al., 1990; Zuardi et al., 2006a,b) effects of
CBD have also further been suggested based on preclinical and
clinical data and it was suggested that CBD may exhibit a
profile similar to atypical antipsychotic drugs (Bhattacharyya
et al., 2009; Borgwardt et al., 2008; Zuardi et al., 2006a,b).
More recently, CBD have also been reported to have anti-oxi-
dative properties (Hampson et al., 1998), which may account to
provide neuroprotection in acute and chronic neurodegenera-
tion reported in different animal models (Garcia-Arencibia et
al., 2007; Lastres-Becker et al., 2005). The anticonvulsivant and
protective effects of CBD against glutamate toxicity may have
a mood stabilizing action similar to some other antiepileptic
drugs of proven value in BD (Ashton and Young, 2003; Porter
et al., 1999).
Previous studies have suggested that oxidative stress may
play a role in the pathophysiology of BD (Andreazza et al.,
2008; Frey et al., 2006b; Machado-Vieira et al., 2007). It has
been demonstrated that valproate and the prototype mood
stabilizer lithium, both first line in the pharmacological treat-
ment of BD, increase brain-derived neurotrophic factor
(BDNF) content in rat hippocampus and frontal cortex.
BDNF is a key regulator of synaptic plasticity and hence is
thought to be uniquely important for neuroprotection.
In addition, it was suggested that these mood stabilizers
exert neuroprotective effects against oxidative stress, indicat-
ing that the regulation of neurotrophic factors might be asso-
ciated with their pharmacological effects.
As the pharmacological profile of CBD has several char-
acteristics in common with drugs known to benefit BD, it
was hypothesized that CBD may have mood stabilizing
properties (Ashton et al., 2005). Therefore, the aim of the
present study was to directly investigate for the first time to
the best of our knowledge, if the administration of CBD can
reverse and/or prevent in rats the behavioral and oxidative
stress effects of chronic use of the indirect dopaminergic
agonist D-amphetamine, in an animal model of mania
(Frey et al., 2006a, b).
Methods
In vivo studies were performed in accordance with National
Institute of Health guidelines and with approval of
Universidade do Extremo Sul Catarinense, Criciu´ ma, SC,
Brazil.
Animals
Male Wistar rats (age, 2–3 months; weight, 250–320 g) were
used in this study. They were housed five to a cage with food
and water available and libitum, and were maintained on a
12 h light/dark cycle (lights on at 7.00 a.m.) in a temperature
controlled (22C) colony room. These conditions were main-
tained constant throughout the experiments.
Drugs
CBD (THC-Pharm, Frankfurt, Germany) was suspended in
polyoxyethylenesorbitan monooleate (Tween 80) 2% saline.
D-AMPH (Sigma, St. Louis, MO, USA) was dissolved in
saline (NaCl 0.9%). The solutions were prepared immediately
before use and were protected from the light during the exper-
imental session.
Reversal treatment
In this model, we reproduced the treatment of an acute manic
episode according to an animal model of mania from
Frey (Frey et al., 2006a). Rats received either a daily injection
of D-amphetamine, 2 mg/kg, or saline for 14 days. Between
the 8th and the 14th days, the animals were divided into four
experimental groups (15 animals per group): CBD (15, 30 or
60 mg/kg) intraperitoneal (i.p.), twice a day, with an interval
of 12 h or saline i.p., twice a day with an interval of 12 h.
Locomotor activity was assessed 2 h after last injection.
Prevention treatment
The second model, we reproduced the maintenance treatment
of BD according to an animal model of mania from
Frey (Frey et al., 2006a). Rats received cannabidiol (15, 30
or 60 mg/kg) or saline i.p. twice a day, in an interval of 12 h
for 14 days. The animals were then divided into two groups
(15 animals per group). Between the 8th and the 14th days,
each group received one daily i.p. injection of D-ampheta-
mine, 2 mg/kg, or saline. Locomotor activity was assessed
2 h after the last injection.
Locomotor activity
We used the open-field task to assess locomotor activity. The
task was performed in a 40 60 cm open field surrounded by
50 cm high walls. The floor of the open field was divided into
12 equal rectangles by black lines. The animals were gently
placed on the left rear rectangle and were allowed to explore
the arena. Crossings of the black lines and rearings
were counted for 5 min (Frey et al., 2006a). The open field
box was cleaned with alcohol 70% among between the
sessions.
Valvassori et al. 275
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Biochemical measures
Measurement of protein carbonyls: The oxidative
damage to proteins was assessed in prefrontal cortex and
hippocampus by the determination of carbonyl groups
based on the reaction with dinitrophenylhydrazine (DNPH)
as previously described (Levine et al., 1990). Briefly, proteins
were precipitated by the addition of 20% trichloroacetic acid
and redissolved in DNPH. The quantification of protein car-
bonyls in the samples was determined in the absorbance of
370 nm. The protein content was normalized by quantifica-
tion according Lowry method (Lowry et al., 1951).
Measurement of BDNF levels: BDNF levels in hippocam-
pus were measured by anti-BDNF sandwich ELISA, according
to the manufacturer instructions (Chemicon, USA). Briefly,
brain slices were homogenized in phosphate-buffered saline
(PBS) with 1 mM phenylmethylsulfonyl fluoride (PMSF) and
1 mM ethylene glycol tetraacetic acid (EGTA). Microtitre
plates (96-well flat-bottom) were coated for 24 h with the sam-
ples diluted 1:2 in sample diluent and standard curve ranged
from 7.8 to 500 pg/ml of BNDF. The plates were then washed
four times with sample diluent and a monoclonal anti-BNDF
rabbit antibody diluted 1:1000 in sample diluent was added to
each well and incubated for 3h at room temperature. After
washing, a peroxidase conjugated anti-rabbit antibody (diluted
1:1000) was added to each well and incubated at room temper-
ature for 1h. After addition of streptavidin enzyme, substrate
and stop solution, the amount of BDNF was determined by
absorbance in 450 nm. The standard curve demonstrates a
direct relationship between optical density (OD) and BDNF
concentration. Total protein was measured by Lowry’s
method using bovine serum albumin as a standard.
Statistical analysis
All data are presented as mean SEM. Differences among
experimental groups in experiment evaluating BDNF levels
were determined by ANOVA. Multiple comparisons were
determined by a Tukey test. In all experiments, p-values
<0.05 were considered to indicate statistical significance.
Results
In the reversal experiment: D-AMPH increased locomotor
and rearing behaviors (Figure 1A and B) in animals treated
with this drug, F¼6.910; p<0.0001 for crossings; F¼7.11,
p<0.0001 for rearings. CBD did not reverse D-AMPH-
induced hyperactivity. The D-AMPH alone administration
increased formation of protein oxidation products in this
treatment in the brain regions analyzed (Figure 1C). In the
prefrontal cortex the D-AMPH-induced damage was
increased with the treatment of CBD 15, 30 or 60 mg/kg. In
the hippocampus CBD 15 mg/kg reversed the D-AMPH-
induced damage, but CBD 30 or 60 mg/kg increased
this damage. In the striatum, no effects in the treatment
with CBD 15 or 30 mg/kg was observed, but CBD 60 mg/kg
increased D-AMPH-induced formation of carbonyl group.
The D-AMPH alone had no effect on BDNF levels in rat
hippocampus (Figure 1D), but CBD 30 mg/kg increased
BDNF expression after AMPH administration. However
CBD 15 or 60 mg/kg had no effect on BDNF levels in
D-AMPH-treated animals. CBD 15, 30 or 60 mg/kg also
had no effect on BDNF levels in saline-treated animals.
In the prevention experiment D-AMPH increased locomo-
tor and rearing Figure 2A, B behavior in animals treated
with this drug, F¼14.63; p<0.0001 for crossings;
F¼8.934; p<0.0001 for rearings. CBD did not reverse D-
AMPH-induced hyperactivity. The D-AMPH alone adminis-
tration increased formation of protein oxidation products in
this treatment in the brains regions analyzed, Figure 2C. The
D-AMPH-induced formation of carbonyl group in the pre-
frontal cortex was prevented by CBD (30 or 60 mg/kg), and
no effect was observed with CBD 15 mg/kg pretreatment. In
the hippocampus and striatum the D-AMPH-induced damage
was prevented by CBD 15, 30 or 60 mg/kg. The D-AMPH
alone had no effect on BDNF levels in rat hippocampus
(Figure 2D). CBD 15, 30 or 60 mg/kg had no effect on
BDNF levels in AMPH- or saline-pretreated animals.
Discussion
In the present study, CBD neither reversed (reversal treat-
ment model) nor prevented (prevention treatment model)
amphetamine-induced hyperactivity in a valid animal model
of mania (Frey et al., 2006a). These data are in line with the
results we have observed in two BD female patients in manic
episodes with psychotic features, who were treated with CBD
for 25 days (initial oral dose of 600 mg reaching 1200mg/day).
Both patients showed no symptoms improvement during
CBD monotherapy with any dose during the trial (Zuardi
et al., 2008a). These preliminary data suggest that CBD
may not be effective for the manic episode of BD.
Nevertheless, in this study we demonstrated that CBD
(15 mg/kg) reversed amphetamine-induced damage and
increased BDNF expression levels (30 mg/kg) after AMPH
administration in rat hippocampus. Moreover, the
D-AMPH-induced damage in the prefrontal cortex was pre-
vented by 30 or 60 mg/kg of CBD and with all doses tested in
the hippocampus and striatum. Interestingly, we have pre-
viously observed that CBD, like clozapine, induced c-Fos
immunoreactivity in prefrontal cortex in rats, distinctively
from haloperidol, that promotes it in dorsal striatum
(Guimara
˜es et al., 2004). Therefore, using the present
model, we were able to reproduce previous findings of the
neuroprotective and antioxidant effects of CBD.
Recently, it was observed that CBD reduces glutamate tox-
icity mediated by N-methyl-D-aspartate receptors (NMDAr),
2-amino-3-(4-butyl-3-hydroxyisoxazol-5-yl) propionic acid
receptors (AMPA) or kainate receptors. This neuroprotection
action of CBD seems to be independent of the CB1 receptor,
the central known cannabinoid receptor, as it has not been
affected by SR-141716A, a CB1 receptor antagonist
(Hampson et al., 1998). Former studies had also demonstrated
that the glutamate toxicity may be prevented by antioxidants
(Cheng et al., 2008; Kuhlmann et al., 2008). Consistent with
this observation, CBD has proven to reduce hydroperoxide-
induced oxidative damage as well as or better than other anti-
oxidants. CBD has shown to be more protective against
276 Journal of Psychopharmacology 25(2)
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glutamate neurotoxicity than the classical antioxidant com-
pounds ascorbate and a-tocopherol (Hampson et al., 1998).
It was hypothesized that the anti-oxidative action of CBD
could be responsible for the neuroprotection reported in
animal models of Parkinson’s disease (PD), as the sub-chronic
administration of CBD reduces toxic effects caused by a uni-
lateral injection of 6-hydroxydopamine into the medial fore-
brain bundle (Lastres-Becker et al., 2005). In this model of PD,
CBD led to an up-regulation of mRNA levels of Cu/Zn-super-
oxide dismutase, a key enzyme in endogenous defense against
oxidative stress. It was concluded that the antioxidant effects of
CBD can provide neuroprotection against the progressive
degeneration of nigrostriatal dopaminergic neurons that
occur in this movement disorder (Garcia-Arencibia et al.,
2007). This observation was corroborated by the fact that
CBD reduced the striatal atrophy caused by 3-nitropropionic
acid, in vivo, through mechanisms independent of the activa-
tion of vanilloid TRPV1, cannabinoid and adenosine A2A
receptors (Sagredo et al., 2007). Using proton magnetic reso-
nance spectroscopy in cannabis users, it was recently found a
strong positive correlation of NAA/tCr and CBD in the puta-
men/globus pallidum and could reflect CBD’s enhancement of
neuronal and axonal integrity in these brain regions (Hermann
et al., 2007). Thus, the prevention of D-AMPH-induced
damage in the striatum with all CBD doses tested observed
in the present study is in line with neuroprotective properties
of CBD, which were also observed in in vitro model studies of
Parkinson’s disease.
Considering the relevance of these preclinical data and the
observed antipsychotic effect of CBD in clinical and preclini-
cal data, we have evaluated, for the first time, the efficacy,
tolerability and safety of CBD in PD patients with psychotic
symptoms. In an open-label pilot study, the PD patients have
shown a significantly decrease both in the psychotic symp-
toms and in the motor function under CBD treatment.
These preliminary data suggests that CBD may be effective
for the treatment of PD (Zuardi et al., 2008b).
The evidences of possible neuroprotective properties of
CBD in both in vitro (Esposito et al., 2006a, b; Iuvone
et al., 2004) and in vivo (Esposito et al., 2007) led to the
importance of studies on the therapeutic potential of this
cannabinoid in Alzheimer’s disease (AD), as this brain disor-
der is strongly related with oxidative stress.
Therefore, considering that the hippocampus neurodegenera-
tion has a key role in AD, the ability of CBD in reversing and
preventing amphetamine-induced damage, and in increasing
BDNF expression levels in hippocampus further highlights
that this compound is very promising to AD prevention.
Control
AMPH+Sal
AMPH+CBD15
AMPH+CBD30
AMPH+CBD60
Control
Sal+CBD15
Sal+CBD30
Sal+CBD60
AMPH+Sal
AMPH+CBD15
AMPH+CBD30
AMPH+CBD60
(B)
∗
∗
∗
∗
0
10
20
30
40
50
60
70
Rearings
(D)
0
10
20
30
40
50
60
70
80
90
Control
Sal+CBD15
Sal+CBD30
Sal+CDB60
AMPH+Sal
AMPH+CBD15
AMPH+CBD30
AMPH+CBD60
pg BDNF/µg of protein
∗
(A)
∗
∗
∗
∗
0
20
40
60
80
100
120
Control
Sal+CBD15
Sal+CBD30
Sal+CBD60
AMPH+Sal
AMPH+CBD15
AMPH+CBD30
AMPH+CBD60
Crossings
(C)
∗
∗
∗
∗
∗/∗∗
∗
∗/∗∗
∗/∗∗
∗/∗∗
∗/∗∗
∗/∗∗
0
2E-12
4E-12
6E-12
8E-12
1E-11
1,2E-11
1,4E-11
1,6E-11
1,8E-11
Prefrontal
Protein carbonyls (nmol/mg protein)
StriatumHippocampus
Figure 1. (A) Number of crossings. (B) Rearings (n¼15 for each group). (C) Protein carbonyl assessment (n¼5 for each group). (D) BDNF levels
(n¼5 for each group) in the reversal model. Rats were pretreated with amphetamine (D-AMPH) for seven days and then treated with amphetamine plus
cannabidiol (15, 30 or 60 mg/kg) between the 8th and 14th days. CBD ¼cannabidiol, control ¼vehicle + saline. Bars represent means; error bars
represent standard error of the means (SEM). *Different to the saline group. **Different to the D-AMPH group.
Valvassori et al. 277
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Interestingly, using functional neuroimaging we have previously
observed that the anxiolytic-like effect induced by CBD is
mediated by an action in the left para-hippocampal gyrus and
left amygdala-hippocampus complex (Crippa et al., 2004).
Despite these observed anti-oxidant findings, in the present
study we have also found that the co-administration of CBD
with amphetamine can increase D-AMPH-induced formation
of carbonyl group, suggesting that their effects on oxidative
stress vary depending on the brain region, treatment and
doses regimen. In fact, these contrasting neuroprotective and
antioxidant CBD effects observed here can be explained by its
multiple mechanisms of action and the fact that many of the
effects of CBD draw a bell-shaped dose–response curve, sug-
gesting that the dose is a key factor in CBD research (Zuardi et
al., 2008b). Moreover, these contrasting results have also been
previously verified with other compounds that exert neuropro-
tective effects such as valproate and lithium (Frey et al., 2006b).
In conclusion, we demonstrated that CBD did not modify
D-AMPH-induced manic-like hyperactivity, but could protect
D-AMPH-induced damage through oxidative stress. In addi-
tion, CBD increased levels of BDNF in the reversal experi-
ment. However, these protective effects depend on the brain
region analyzed, treatment and doses regimen. Our findings
further support the notion that CBD may have neuroprotec-
tive effects, although more research is still needed to clarify its
precise mechanisms that underlie this potentially beneficial
effect of CBD.
Acknowledgements
This study was supported in part by grants from ‘Conselho Nacional
de Desenvolvimento Cientı
´fico e Tecnolo
´gico’ (CNPq-Brazil-554490/
2005–6), ‘Fundac¸ a
˜odeAmparoa
`Pesquisa do Estado de Sa
˜oPaulo
fellowship’ (FAPESP -02/13197–2), ‘Instituto Ce
´rebro e Mente’ (FK,
FDP, and JQ), UNESC (FDP, and JQ) and FAPESC (FDP, and JQ).
This study was also sponsored by THC-Pharm (Frankfurt, Germany)
and STI Pharmaceuticals Ltd, (Brentwood, UK) who kindly provided
cannabidiol). JAC, AWZ, JQ, FDP and FK are recipients of a CNPq
Productivity fellowship. SSV is holder of a CAPES studentship; CT is
holder of a FAPESP studentship and GE is holder of a UNESC
studentship.
References
Andreazza AC, Kauer-Sant’anna M, Frey BN, et al. (2008) Oxidative
stress markers in bipolar disorder: a meta-analysis. J Affect
Disord 111: 135–144.
(A)
0
20
40
60
80
100
120
Control
CBD15+Sal
CBD30+Sal
CBD60+Sal
Sal+AMPH
CBD15+AMPH
CBD30+AMPH
CBD60+AMPH
Crossings
∗
∗
∗∗
(B)
0
10
20
30
40
50
60
70
Control
CBD15+Sal
CBD30+Sal
CBD60+Sal
Sal+AMPH
CBD15+AMPH
CBD30+AMPH
CBD60+AMPH
Rearings
∗
∗
∗
∗
(C)
∗
∗
l
∗
∗∗
∗/∗∗
∗/∗∗
∗/∗∗
∗/∗∗
∗/∗∗
0
2E-12
4E-12
6E-12
8E-12
1E-11
1,2E-11
Prefrontal
Protein carbonyl (nmol/mg protein)
Control
AMPH+Sal
AMPH+CBD15
AMPH+CBD30
AMPH+CBD60
(D)
0
10
20
30
40
50
60
70
Control
CBD15+Sal
Sal+CBD30
CDB60+Sal
Sal+AMPH
CBD15+AMPH
CBD30+AMPH
CBD60+AMPH
pg BDNF/µg of protein
StriatumHippocampus
Figure 2. (A) Number of crossings. (B) Rearings (n¼15 for each group). (C) Protein carbonyl assessment (n¼5 for each group). (D) BDNF levels
(n¼5 for each group) in the prevention model. Rats were pretreated with cannabidiol (15, 30 or 60 mg/kg) for seven days and then treated with
amphetamine plus amphetamine (D-AMPH) the 8th and 14th days. CBD ¼cannabidiol, control ¼vehicle+saline. Bars represent means; error bars
represent standard error of the means (SEM). *Different to the saline group. **Different to the D-AMPH group.
278 Journal of Psychopharmacology 25(2)
at UNIV EXTREMO SUL CATARINENSE P on January 23, 2015jop.sagepub.comDownloaded from
Ashton H, Young AH (2003) GABA-ergic drugs: exit stage left, enter
stage right. J Psychopharmacol 17: 174–178.
Bhattacharyya S, Crippa JA, Martin-Santos R, Winton-Brown T,
Fusar-Poli P (2009) Imaging the neural effects of cannabinoids:
current status and future opportunities for psychopharmacology.
Curr Pharm Des in press.
Borgwardt SJ, Allen P, Bhattacharyya S, et al. (2008) Neural basis of
delta-9-tetrahydrocannabinol and cannabidiol: effects during
response inhibition. Biol Psychiatry 64: 966–973.
Carlini EA, Leite JR, Tanhauser M, Berardi AC (1973) Cannabidiol
and cannabis sativa extract protect mice and rats against convul-
sive agents. J Pharm Pharmacol 25: 664–665.
Cheng HY, Hsieh MT, Wu CR, et al. (2008) Schizandrin protects
primary cultures of rat cortical cells from glutamate-induced exci-
totoxicity. J Pharmacol Sci 107: 21–31.
Crippa JA, Zuardi AW, Garrido GE, et al. (2004) Effects of canna-
bidiol (CBD) on regional cerebral blood flow.
Neuropsychopharmacology 29: 417–426.
Cunha JM, Carlini EA, Pereira AE, et al. (1980) Chronic adminis-
tration of cannabidiol to healthy volunteers and epileptic patients.
Pharmacology 21: 175–185.
Esposito G, De Filippis D, Carnuccio R, Izzo AA, Iuvone T (2006a)
The marijuana component cannabidiol inhibits beta-amyloid-
induced tau protein hyperphosphorylation through Wnt/beta-
catenin pathway rescue in PC12 cells. J Mol Med 84: 253–258.
Esposito G, De Filippis D, Maiuri MC, De Stefano D, Carnuccio R,
Iuvone T (2006b) Cannabidiol inhibits inducible nitric oxide
synthase protein expression and nitric oxide production in beta-
amyloid stimulated PC12 neurons through p38 MAP kinase and
NF-kappaB involvement. Neurosci Lett 399: 91–95.
Esposito G, Scuderi C, Savani C, et al. (2007) Cannabidiol in vivo
blunts beta-amyloid induced neuroinflammation by suppressing
IL-1beta and iNOS expression. Br J Pharmacol 151: 1272–1279.
Frey BN, Andreazza AC, Cerese
´r KM, et al. (2006a) Effects of mood
stabilizers on hippocampus BDNF levels in an animal model of
mania. Life Sci 79: 281–286.
Frey BN, Valvassori SS, Re
´us GZ, et al. (2006b) Effects of lithium and
valproate on amphetamine-induced oxidative stress generation in
an animal model of mania. J Psychiatry Neurosci 31: 326–332.
Fusar-Poli P, Crippa JA, Bhattacharyya S, et al. (2009) Neurofunctional
modulation of emotional processing by 9-tetrahydrocannabinol
and cannabidiol. Arch Gen Psychiatry 66: 95–105.
Garcia-Arencibia M, Gonzalez S, de Lago E, Ramos JA, Mechoulam
R, Fernandez-Ruiz J (2007) Evaluation of the neuroprotective
effect of cannabinoids in a rat model of Parkinson’s disease:
importance of antioxidant and cannabinoid receptor-independent
properties. Brain Res 1134: 162–170.
Goodwin FK (2003) Impact of formularies on clinical innovation.
J Clin Psychiatry 64(Suppl 5): 18–24.
Guimara
˜es FS, Chiaretti TM, Graeff F, Zuardi AW (1990)
Antianxiety effect of cannabidiol in the elevated plus-maze.
Psychopharmacology (Berl) 100: 558–559.
Guimara
˜es VM, Zuardi AW, Del Bel EA, Guimara
˜es FS (2004)
Cannabidiol increases Fos expression in the nucleus accumbens
but not in the dorsal striatum. Life Sciences 75: 633–638.
Hamelink C, Hampson A, Wink DA, Eiden LE, Eskay RL (2005)
Comparison of cannabidiol, antioxidants, and diuretics in rever-
sing binge ethanol-induced neurotoxicity. J Pharmacol Exp Ther
314: 780–788.
Hampson AJ, Grimaldi M, Axelrod, Wink D (1998) Cannabidiol and
delta 9-tetrahydrocannabinol are neuroprotective antioxidants.
Proc Natl Acad Sci USA 95: 8268–8273.
Hermann D, Sartorius A, Welzel H, et al. (2007) Dorsolateral pre-
frontal cortex N-acetylaspartate/total creatine (NAA/tCr) loss in
male recreational cannabis users. Biol Psychiatry 61: 1281–1289.
Iuvone T, Esposito G, Esposito R, Santamaria R, Di Rosa M, Izzo
AA (2004) Neuroprotective effect of cannabidiol, a
non-psychoactive component from Cannabis sativa, on beta-amy-
loid-induced toxicity in PC12 cells. J Neurochem 89: 134–141.
Izquierdo I, Orsingher OA, Berardi AC (1973) Effect of cannabidiol
and other Cannabis sativa compounds on hippocampal seizures
discharges. Psychopharmacologia 28: 95–102.
Kuhlmann CR, Gerigk M, Bender B, Closhen D, Lessmann V,
Luhmann HJ (2008) Fluvastatin prevents glutamate-induced
blood–brain-barrier disruption in vitro. Life Sci 8: 1281–1287.
Lastres-Becker I, Molina-Holgado F, Ramos JA, Mechoulam R,
Fernandez-Ruiz J (2005) Cannabinoids provide neuroprotection
against 6-hydroxydopamine toxicity in vivo and in vitro: rele-
vance to Parkinson’s disease. Neurobiol Dis 19: 96–107.
Machado-Vieira R, Andreazza AC, Viale CI, et al. (2007) Oxidative
stress parameters in unmedicated and treated bipolar subjects
during initial manic episode: a possible role for lithium antioxi-
dant effects. Neurosci Lett 421: 33–36.
Mitchell PB, Malhi GS (2006) Emerging drugs for bipolar disorder.
Expert Opin Emerg Drugs 11: 621–634.
Monti JM (1977) Hypnotic-like effects of cannabidiol in the rat.
Psychopharmacol (Berl) 55: 263–265.
Moreira FA, Aguiar DC, Guimara
˜es FS (2006) Anxiolytic-like effect
of cannabidiol in the rat Vogel conflict test. Prog
Neuropsychopharmacol Biol Psychiatry 30: 1466–1471.
Musty RE, Bouldin MJ, Erickson JR, Deyo R (2002) CBD and THC
extracts alter Behavioral Despair on the Mouse Tail Suspension
Test. Asilomar Conference Center. 2002 Symposium on the
Cannabinoids, Burlington, VT. International Cannabinoid
Research Society, p. 140.
Onaivi ES, Green MR, Martin BR (1990) Pharmacological charac-
terization of cannabinoids in the elevated plus maze. J Pharmacol
Exp Ther 253: 1002–1009.
Porter R, Ferrier N, Ashton H (1999) Anticonvulsants as mood sta-
bilisers. Adv Psychiatr Treat 5: 96–103.
Sagredo O, Ramos JA, Decio A, Mechoulam R, Ferna
´ndez-Ruiz J
(2007) Cannabidiol reduced the striatal atrophy caused 3-nitro-
propionic acid in vivo by mechanisms independent of the activa-
tion of cannabinoid, vanilloid TRPV1 and adenosine A2A
receptors. Eur J Neurosci 26: 843–851.
Turkanis SA, Cely W, Olsen DM, Karler R (1974) Anticonvulsant
properties of cannabidiol. Res Communic Chem Pathol Pharmacol
8: 231–246.
Zuardi AW (2006) History of cannabis as a medicine: a review. Rev
Bras Psiquiatr 28: 153–157.
Zuardi AW, Cosme RA, Graeff FG, et al. (1993) Effects of
ipsapirone and cannabidiol on human experimental anxiety.
J Psychopharmacol 7: 82–88.
Zuardi AW, Crippa JAS, Dursun SM, et al. (2008a) Cannabidiol was
not effective for manic episode of bipolar affective disorder.
J Psychopharmacol doi:10.1177/0269881108096521, in press.
Zuardi AW, Crippa JAS, Hallak JEC, et al. (2008b) Cannabidiol
for the treatment of psychosis in Parkinson’s disease.
J Psychopharmacology 23: 979–983.
Zuardi AW, Crippa JA, Hallak JE, Moreira FA, Guimara
˜es FS
(2006a) Cannabidiol, a Cannabis sativa constituent, as an antipsy-
chotic drug. Braz J Med Biol Res 39: 421–429.
Zuardi AW, Hallak JE, Dursun SM, et al. (2006b) Cannabidiol
monotherapy for treatment-resistant schizophrenia.
J Psychopharmacol 20: 683–686.
Zuardi AW, Karniol IG (1983) Effects on variable-interval perfor-
mance in rats of delta 9-tetrahydrocannabinol and cannabidiol,
separately and in combination. Braz J Med Biol Res 16: 141–146.
Zuardi AW, Shirakawa I, Finkelfarb E, Karniol IG (1982). Action of
cannabidiol on the anxiety and other effects produced by delta 9-
THC in normal subjects. Psychopharmacology 76: 245–250.
Valvassori et al. 279
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