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Cocaine is a commonly abused illicit drug that causes significant morbidity and mortality. The most severe and common complications are seizures, ischemic strokes, myocardial infarction, and acute liver injury. Here, we demonstrated that acute cocaine intoxication promoted seizure along with acute liver damage in mice, with intense inflammatory infiltrate. Considering the protective role of the endocannabinoid system against cell toxicity, we hypothesized that treatment with an anandamide hydrolysis inhibitor, URB597, or with a phytocannabinoid, cannabidiol (CBD), protects against cocaine toxicity. URB597 (1.0 mg/kg) abolished cocaine-induced seizure, yet it did not protect against acute liver injury. Using confocal liver intravital microscopy, we observed that CBD (30 mg/kg) reduced acute liver inflammation and damage induced by cocaine and prevented associated seizure. Additionally, we showed that previous liver damage induced by another hepatotoxic drug (acetaminophen) increased seizure and lethality induced by cocaine intoxication, linking hepatotoxicity to seizure dynamics. These findings suggest that activation of cannabinoid system may have protective actions on both liver and brain induced by cocaine, minimizing inflammatory injury promoted by cocaine, supporting its further clinical application in the treatment of cocaine abuse.
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Research Article
Cannabidiol Rescues Acute Hepatic Toxicity and
Seizure Induced by Cocaine
Luciano Rezende Vilela,1,2 Lindisley Ferreira Gomides,1
Bruna Araújo David,1Maísa Mota Antunes,1Ariane Barros Diniz,1
Fabrício de Araújo Moreira,2and Gustavo Batista Menezes1
1Laborat´
orio de Imunobiofotˆ
onica, Departamento de Morfologia, Instituto de Ciˆ
encias Biol´
ogicas,
Universidade Federal de Minas Gerais, Avenida Presidente Antˆ
onio Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil
2Departamento de Farmacologia, Instituto de Ciˆ
encias Biol´
ogicas, Universidade Federal de Minas Gerais,
Avenida Presidente Antˆ
onio Carlos 6627, 31270-901 Belo Horizonte, MG, Brazil
Correspondence should be addressed to Fabr´
ıcio de Ara´
ujo Moreira; farmoreira@yahoo.com.br and
Gustavo Batista Menezes; menezesgb@gmail.com
Received  January ; Accepted  April 
Academic Editor: Donna-Marie McCaerty
Copyright ©  Luciano Rezende Vilela et al. is is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly
cited.
Cocaine is a commonly abused illicit drug that causes signicant morbidity and mortality. e most severe and common
complications are seizures, ischemic strokes, myocardial infarction, and acute liver injury. Here, we demonstrated that acute
cocaine intoxication promoted seizure along with acute liver damage in mice, with intense inammatory inltrate. Considering the
protective role of the endocannabinoid system against cell toxicity, we hypothesized that treatment with an anandamide hydrolysis
inhibitor, URB, or with a phytocannabinoid, cannabidiol (CBD), protects against cocaine toxicity. URB (. mg/kg) abolished
cocaine-induced seizure, yet it did not protect against acute liver injury. Using confocal liver intravital microscopy, we observed that
CBD ( mg/kg) reduced acute liver inammation and damage induced by cocaine and prevented associated seizure. Additionally,
we showed that previous liver damage induced by another hepatotoxic drug (acetaminophen) increased seizure and lethality
induced by cocaine intoxication, linking hepatotoxicity to seizure dynamics. ese ndings suggest that activation of cannabinoid
system may have protective actions on both liver and brain induced by cocaine, minimizing inammatory injury promoted by
cocaine, supporting its further clinical application in the treatment of cocaine abuse.
1. Introduction
Drug abuse and addiction constitute a public health problem
of great importance with a high prevalence worldwide.
AccordingtotheUnitedNationsOceonDrugsandCrime
(UNODC), cocaine, a psychostimulant and psychotomimetic
drug, is among the most abused drugs in the world. e
estimated number of cocaine users globally ranges between
 and  million (.–.% of the population aged between
 and  years) []. Besides its toxicity for the cardiovascular
central nervous systems, cocaine causes liver injury in human
and animal models [,]. is drug may lead to severe
acute hepatotoxicity due to hepatocellular necrosis, which
can be life threatening. Indeed, as diminished liver function
contributes to various adverse health eects, hepatotoxicity
has been linked to the mortality in cocaine abusers [].
Oxidative stress (OS) plays a key role in cocaine-induced
hepatotoxicity and is well reported in both humans and ani-
mal models []. Furthermore, several lines of evidence indi-
cate that excessive OS with increased free radical generation,
impairment of mitochondrial respiration, and intense inam-
matory reaction plays a crucial role in cocaine-induced hepa-
totoxicity. OS is associated with the dysregulation of signaling
pathways modulating cell death and survival, which leads to
necrosis and apoptosis of hepatocytes []. Although previous
studies have yielded mechanistic information on cocaine-
induced hepatotoxicity, the current understanding on this
toxic event remains insucient []. Cocaine users have
Hindawi Publishing Corporation
Mediators of Inflammation
Volume 2015, Article ID 523418, 12 pages
http://dx.doi.org/10.1155/2015/523418
Mediators of Inammation
altered levels of proinammatory cytokines and chemokines
in the plasma, indicating that peripheral immune system
adaptations may contribute to the eects of cocaine [,].
It is well described that the prevention of OS generation
might have therapeutic advantages in several diseases [,,
]; therefore, the investigation of putative new compounds
with antioxidant properties might be promising. e herb
Cannabis sativa (“marijuana,” “hemp”) has been known
for centuries due to its abuse-related eects and possible
therapeutic uses, including the treatment of inammation,
diabetes, cancer, aective or neurodegenerative diseases, and
epilepsy-related disorders. Among more than  chemical
compounds already characterized from this herb, the two
most investigated are Δ9-tetrahydrocannabinol (Δ9-THC)
and cannabidiol (CBD) []. Δ9-THC, which accounts
formostofthetypicalcannabisactions,mayhavethera-
peutic ecacy against several diseases, although its use is
limitedbyitspsychoactiveproperties.us,analternative
for developing cannabis-based medicines could be focusing
on CBD, the major nonpsychotomimetic phytocannabinoid.
is compound acts through multiple pharmacological tar-
gets, including facilitation of the endocannabinoid system,
activation of transient receptor potential vanilloid type-
 (TRPV) channel, the peroxisome proliferator-activated
receptor 𝛾(PPAR𝛾), GPR, -hydroxytryptamine receptor
subtype A (-HT1A), the adenosine membrane transporter
phospholipase A2, lipoxygenase (LOX) and cyclooxygenase-
 (COX-) enzymes, and Ca2+ homeostasis []. is rich
pharmacology, associated with the fact that it does not
share the psychotomimetic, amnestic, and sedative eects
of Δ9-THC, results in a wide range of potential thera-
peutic interest, including the treatment of epilepsies and
related disorders [,]. Accordingly, a recent clinical
study observed that CBD-enriched cannabis extracts signi-
cantly reduced seizure frequency in children with treatment-
resistant epilepsy, indicating that it could be used as a strategy
when other pharmacological treatments fail [].
Apart from searching for the therapeutic application
of phytocannabinoids, other potential strategy focuses on
the endocannabinoid system. is intercellular message
system comprises the CB1and CB2cannabinoid receptor,
the ligands arachidonoyl ethanolamide (anandamide) and
-arachidonoyl glycerol, their hydrolyzing enzymes (fatty
acid amide hydrolase, FAAH, and monoacyl glycerol lipase,
resp.), and their synthesizing machinery [,]. Among its
numerous biological functions, the endocannabinoid system
has been implicated in protection against deleterious stimuli
in various tissues, including the brain []andtheliver[,
]. Previous studies also demonstrated that CBD possesses
ecient antioxidant and anti-inammatory activities against
ischemia/reperfusion liver injury in rats [,]. Moreover,
CBD restored liver/brain function in a model of hepatic
encephalopathy associated with fulminant hepatic failure
induced in mice revealing that its eects may result from a
combination of itsactions in the liver and brain []. Here, to
investigate if CBD exerts protective by CBD exerts protective
eects in a model of seizures and liver damage resulting from
cocaine intoxication. We have compared the eects of these
phytocannabinoids with the FAAH inhibitor URB, which
facilitate endocannabinoid signaling by selectively increasing
anandamidelevels.Finally,weinvestigatedifprevioushepatic
lesion induced by other compounds (acetaminophen) [
] would modulate SNC-related cocaine eects.
2. Materials and Methods
2.1. Mice. Male Swiss mice weighing – g from the animal
facility (Centro de Bioterismo, CEBIO) of UFMG were kept
on a h:h dark/light cycle at 22 ± 1Cwithfreeaccess
to food and water throughout the experiment. Each animal
was used only once. All experiments were conducted in
accordance with the Ethical Committee for Animal Exper-
imentation (CETEA) of the Federal University of Minas
Gerais (Universidade Federal de Minas Gerais (UFMG)) and
procedures for animal care were previously approved by this
organization under Protocol /.
2.2. Drugs. Cannabidiol ( mg/kg; THC-Pharm, Frankfurt,
Germany, and STI-Pharm, Brentwood, UK) was dissolved in
physiological saline containing tween- at %. URB
(. mg/kg; Tocris) was dissolved in ethanol/cremophor/
saline  :  : . Cocaine ( mg/kg; Merck, Darmstadt, Ger-
many) was dissolved in physiological saline. e solutions
were prepared immediately before use and injected via
intraperitoneal route in a volume of  mL/kg. All doses
were chosen based on previously published studies [
]. Cannabidiol dose was chosen based on dose-response
curves previously established in our group []. Although
most of these studies have focused on the anxiolytic and
antipsychotic eects of this compound, the dose range for its
antiseizure and antiepileptic eects is in the sane range [].
us, we tested anticonvulsant eect of cannabidiol (, ,
and  mg/kg)  minutes before cocaine administration. At
dose of  mg/kg, behavioral parameters of seizure (latency
and duration seizure) are signicantly inhibited when com-
pared to doses of  and  mg/kg (data not shown).
2.3. Apparatus and Cocaine-Induced Seizures. e animals
were injected intraperitoneally (i.p.) with cocaine ( mg/kg)
and immediately placed in individual chambers ( cm in
diameter with a  cm high Plexiglas wall) and observed
during  min for the onset of behavior seizures and the
occurrence of death. Seizure was dened as the occurrence
of tail clonus with myoclonic jerks and wild jumping or
convulsions with loss of righting reex. Latency and duration
for myoclonic seizure were also measured in the subset of
mice that progressed to this state.
2.4. Model of Acetaminophen-Induced Liver Injury and Inam-
mation. Acetaminophen (paracetamol; APAP) was orally
administered ( or  mg/kg; Sigma) in male Swiss
mice weighing – g from the animal facility (Centro de
Bioterismo, CEBIO) of UFMG aer  h of fasting [].
Control mice received warm sterile saline as vehicle. Aer
 h, mice were anesthetized with a mixture of ketamine
( mg/kg) and xylazine ( mg/kg) and killed for serum and
Mediators of Inammation
liver harvesting. In a separate set of experiments, the survival
percentageofAPAP-challengedmicewasevaluatedduring
 h. Neutrophil inltration into the liver was measured by
the myeloperoxidase (MPO) activity assay.
2.5. Alanine Aminotransferase Assay (ALT). e alanine
aminotransferase enzyme is present in the cytoplasm of hep-
atocytes and is highly specic for the liver. e measurement
of serum alanine aminotransferase (ALT) is a gold-standard
marker of liver damage. To determine the activity of ALT,
blood samples were centrifuged and the serum was collected
and dosed using a kinetic kit (Bioclin, Brazil) [,].
2.6. Liver Histological Analysis. Livers were collected  h
aer induction of ALF and xed by overnight immersion in
% buered formalin. Paran-embedded specimens were
prepared and . 𝜇m sections were stained with hematoxylin
and eosin (H&E) according to a standard protocol. Liver
pathology was assessed by an investigator who was blinded
to the experimental treatment groups in an Olympus BX
microscope (Center Valley, PA, USA) [,].
2.7. Indocyanine Green (Cardiogreen; ICG). e ICG stan-
dard curve was drawn according to the manufacturer’s
instructions. Indocyanine green was completely dissolved
in distilled water and prepared to a nal concentration of
. mg/mL. According to the experimental design, each ani-
mal was given a caudal intravenous injection of indocyanine
green at  mg/kg of body weight. One hour aer the ICG
injection, a laparotomy was performed and the aorta was
cut, followed by isolation of plasma with centrifugation at
 rpm for  min. Plasma (. mL) was extracted and
diluted with saline. Absorbance value of the plasma sam-
ple was determined by spectrophotometry determining the
absorbance of the plasma sample at nm, which was
compared with the value for normal plasma. e standard
curve was used to calculate the serum concentration of ICG.
2.8. Liver Confocal Intravital Microscopy. Liver microcircula-
tion was imaged as described previously [,]. Briey, mice
were anesthetized with a mixture of ketamine ( mg/kg)
and xylazine ( mg/kg). e liver was gently pulled out
through a laparotomy incision and positioned over a Plexiglas
stage for imaging under a confocal inverted microscope
(Nikon Eclipse Ti and C confocal head). For imaging liver
microcirculation and necrosis, mice received endovenously
𝜇g of phycoerythrin-labeled anti-PECAM- (stains liver
sinusoidal endothelial cells; eBiosciences, USA) and 𝜇Lof
the stock solution of Sytox Green (stains extracellular DNA
deposits and necrosis; Life Technologies, USA). e liver
surface was imaged in a custom made stage that holds the
liver in at position. Fluorophores were excited with 
andnmlaserslineusingaxobjective.Allimageswere
generated by Volocity soware (.; PerkinElmer, USA).
2.9. Statistical Analysis. Experimental data analysis was per-
formed with one-way analysis of variance (ANOVA) (Tukey’s
post hoc test). 𝑃values <. were considered statistically
signicant. All experiments included 𝑛=6.Dataare
presented as mean ±SEM. Fisher’s exact probability test was
additionally used for specic comparisons between seizure
parameters and hepatic functions. Graphs and statistical
analysis were performed using Prism  (GraphPad soware,
USA).
3. Results
3.1. Cocaine Administration Caused Acute Liver Injury and
Inammation. To investigate the hepatotoxic eects of acute
cocaine intake, mice received a single administration of
cocaine ( mg/Kg; i.p.) and livers were imaged under
confocal intravital microscopy. As shown in Figure (a),
control mice had a well-perfused liver microvasculature, as
evidenced by sinusoidal staining by anti-CD (PECAM-
, in red). Also, intravenous administration of a DNA-
binding probe (Sytox green) revealed almost no extracellular
DNA under basal conditions. We have previously shown
that hepatocytes released DNA into the liver, leading to a
widespread hepatic DNA accumulation, which was directly
correlated with injury severity and progression. In line
with this, mice overdosed with cocaine had massive liver
injury, evidenced by lack of perfusion (dark spots on red
channel), which were consistently lled by extracellular DNA,
linking cocaine intake to acute liver injury. Elevated serum
transaminases levels (ALT) conrmed acute hepatotoxicity
induced by cocaine (Figure (b)). To evaluate if such injury
was ultimately impairing liver function, we injected a known
amount of indocyanine green (cardiogreen) [], which is
rapidly cleared from the circulation under control conditions
by the liver. Interestingly, cocaine-overdosed mice presented
a signicant retention of cardiogreen in the serum, suggesting
that liver metabolic function was also impaired (Figure (c)).
In fact, liver histopathology showed that cocaine also caused
liver leukocyte inltration together with an extensive necrosis
(Figures (a) and (b)). Together, these data demonstrated
that acute cocaine intake led to liver injury and inammation.
3.2. Cocaine-Induced Seizure Is Directly Correlated with Liver
Injur y. In order to elucidate the relationship between liver
injury and seizures, we rst studied the dynamics of seizure
due to cocaine overdose. Following a single cocaine injection,
seizure was observed in all animals, manifested as rapid
myoclonic convulsions of forelimbs and tail clonus preceded
by locomotor depression (Figure (a)), as demonstrated by
reduced seizure latency and long lasting seizure (Figure (b)).
Interestingly, we observed a direct correlation between liver
injury and function with seizure duration, suggesting that
reducedlivermetabolismmightpredisposetomoresevere
seizureduetococaineintake(Figure (c)).isisparticularly
relevant in the clinics because it might suggest that patients
with previous liver damage are more susceptible to convulsive
seizure during cocaine overdose. In this direction, we next
investigated if a previous liver injury caused by another
hepatotoxic drug worsens seizure prole induced by cocaine.
For this, we established a model of hepatic injury due to
acetaminophen (APAP;  mg/Kg) overdose. As shown in
Mediators of Inammation
PECAM-1
(sinusoids)
DNA (necrosis) Merge
Vehicle
Cocaine
(a)
0
1000
2000
3000
ALT (U/L)
Veh i c l e Cocaine
(b)
Veh i c l e Cocaine
0
10
20
30
40
50
Cardiogreen (𝜇g/mL)
(c)
F : Cocaine intake causes severe liver injury and malfunction. (a) Liver confocal intravital microscopy showing that while control mice
have a well perfused liver vasculature (evidenced by PE-anti-CD antibody staining, in red) and absence of necrosis (in green), cocaine-
treated mice had several dark areas in the red channel, which are suggestive of malperfusion caused by necrosis (elicited by DNA staining
using Sytox Green). (b) Serum liver transaminase levels and (c) retention of cardiogreen conrmed necrosis. Statistical signicance in
comparison to controls (𝑃 < 0.05). A x objective was used during imaging. One-way ANOVA with Tukey’s post hoc test. Data are expressed
as mean ±S.E.M. (𝑛=6mice/treatment).
Figure (a), APAP caused extensive liver injury following
 hs of overdose, and overt DNA deposition (in green)
wasobservedinnonperfusedareas(blackareasinthered
channel). Elevated levels of serum ALT (Figure (b))and
histopathology examination conrmed acute liver injury
(Figure (a)), which occurred concomitantly to hepatic neu-
trophil inltration (Figure (b)). APAP-treated mice that
werechallengedwithasingledoseofcocainehadsig-
nicantly higher lethality in comparison to cocaine alone
(Figure (a)). However, mice treated with higher doses of
APAP had lower seizure incidence (Figure (b))andduration
(Figure (c)), with higher latency (Figure (d)), suggesting
that metabolic brain disorders induced by acute liver failure
can interfere in seizure dynamics, reinforcing the direct
correlation of liver metabolic status with brain function.
3.3. Pretreatment with Cannabidiol (CBD) Reduces Cocaine-
Induced Liver Injur y and Seizure. It is well accepted that
cannabinoids may have anti-inammatory activities and had
been proposed to treat neurological disorders. In this context,
the use of nonpsychotropic molecules is preferable in com-
parison to the herb itself, which also oers the opportunity
to dosage adjustments. Taking into account the relationship
between hepatic inammation and brain disorders, we next
investigated the participation of the cannabinoid system in
our model. For this, we pretreated mice with cannabidiol
(CBD;  mg/Kg), a major nonpsychotomimetic phytocan-
nabinoid. In fact, pretreatment with CBD completely rescued
liver injury induced by cocaine, as assessed by reduced
liver DNA deposition (Figure (a)), and reduced serum
ALT (Figure (b))andcardiogreen(Figure (c))levels,
Mediators of Inammation
Cocaine
Veh i c l e
(a)
0
10
20
30
40
50
*
Necrosis area (%)
Veh i c l e C o cain e
0
1
2
3
Histological score
Veh i c l e C o cain e
0
50
100
150
200
Veh i c l e Cocaine
Neutrophils (×104)
(b)
F : Histopathological assessment of cocaine-induced liver injury. (a) Liver histology (H&E) showing that while control mice have a
healthy liver architecture and absence of necrosis, cocaine-treated mice had several necrotic areas with leukocyte inltration (green arrow). (b)
Digital quantication of necrotic areas using ImageJ, histopathological score, and neutrophil inltration (assessed by MPO levels) conrmed
severe liver inammation and necrosis induced by cocaine. Statistical signicance in comparison to controls (𝑃 < 0.05). A x objective was
used during imaging. One-way ANOVA with Tukey’s post hoc test. Data are expressed as mean ±S.E.M(𝑛=6mice/treatment).
which prevented hepatic dysfunction due to cocaine over-
dose. Histopathology analysis conrmed that CDB-treated
mice had no detectable injury due to cocaine injection
(Figure (a)), and lower liver inammation (estimated by
neutrophil inltration) was also observed (Figure (b)). In
line with this, CBD treatment also reduced cocaine-induced
seizure duration and higher latency (Figure (c)). Taking into
account the potent eects of CBD, we next investigated if the
inhibition of degradation of endogenous endocannabinoids
might mimic the benecial eects of the administration
of the exogenous compound (CBD). URB mimicked
the anticonvulsant eects of CBD, and URB-treated mice
had less severe seizure induced by cocaine (Figure (c)).
However, pretreatment with URB caused no detectable eects
of cocaine-induced liver injury, suggesting that despite its
protective central eects on seizure dynamics, exogenous
agonists and higher doses are necessary for reaching the
hepatoprotective eects of cannabinoid system activation.
4. Discussion
In the present study we demonstrated that CBD inhibited
cocaine-induced seizure and liver injury in mice, alleviating
hepatic inammatory process, and reduced cocaine lethality.
e FAAH inhibitor, URB, inhibited seizure but did
not interfere with hepatic inammatory process. We also
observed that previous hepatic dysfunction caused by APAP
increased cocaine-induced seizure and lethality.
Drugabuseandaddictionconstituteapublichealthprob-
lem of great importance with a high prevalence worldwide.
AccordingtotheUnitedNationsOceonDrugsandCrime
(UNODC, ), estimated number of cocaine users globally
ranges between  and  million (.–.% of the population
aged between  and  years) []. Acute cocaine abuse may
induce several neurological impairments, including seizures
and fulminant hepatic failure. Since there are no eective
treatments available, it is associated with severe intoxications
Mediators of Inammation
(a)
0
25
50
75
100
*
Seizure (%)
Veh i c l e Cocaine
0
200
400
600
800
Total experimental time
Seizure latency (s)
Veh i c l e C o cain e
0
10
20
30
40
50
Seizure duration (s)
Veh i c l e C o cain e
(b)
0 20406080
0
500
1000
1500
2000
2500
3000
3500
4000
Seizure duration
ALT (U/L)
r2= 0.86
0204060
30
35
40
45
50
Seizure duration
Cardiogreen (𝜇g/mL)
r2= 0.74
(c)
F : High doses of cocaine promote acute behavioral seizure. (a) Representative picture of seizure characterized by rapid myoclonic
convulsions of forelimbs and tail clonus preceded by locomotor depression. (b) Acute overdose induced seizure in all animals represented by
latency reduced and duration increased compared to group vehicle. (c) Positive correlation between seizure duration and acute hepatotoxicity
induced by cocaine (𝑟2= 0.86,𝑃 < 0.02for ALT; 𝑟2= 0.74,𝑃 < 0.05for cardiogreen). Signicantly dierent from vehicle group (𝑃< 0.001).
One-way ANOVA with Tukey’s post hoc test. Data are expressed as mean ±S.E.M (𝑛=6mice/treatment).
and deaths [,]. us, our results showed that cannabidiol
displays neuroprotective and hepatoprotective eects against
cocaine toxicity and therefore might be used as future strategy
in clinical emergency. Although our data provide support for
the ecacy of CBD in this condition, careful pharmacologic
studies are needed to further delineate specic mechanisms
of CBD against cocaine toxicity.
e mechanisms involved in cocaine intoxication and
seizure induction have remained unclear. Its psychotomi-
metic and psychostimulant eects are mediated by blockade
of dopamine, serotonin, and norepinephrine reuptake. e
increased stimulation of dopamine D receptors may also
contribute to cocaine-induced seizures []. Clinical []
and experimental [,] observations demonstrate that
the acute administration of high doses of this drug promotes
convulsive seizures and liver abnormalities or fulminant
hepatic failure. Less attention, however, has been directed
towards possible hepatotoxic eects []. is drug induces
increase in ALT activity and pervasive centrilobular foci of
necrosis crowded with pale and swollen hepatocytes [].
Several case reports have described hepatic injury from
cocaine use, including cases in which cocaine-induced liver
damage resulted in mortality [].
Here, we described for the rst time that previous treat-
ment with phytocannabinoid CBD abolished cocaine in-
duced acute seizure in mice, indicating anticonvulsant eect,
in parallel with anti-inammatory eect and protection
against liver injury. CBD is the major nonpsychoactive com-
ponent of cannabis that exerts multiple pharmacological
actions in the central nervous system and in the periphery,
Mediators of Inammation
PECAM-1
(sinusoids)
DNA (necrosis) Merge
Vehicle
APAP
(a)
0
1000
2000
3000
ALT (U/L)
Vehicle APAP
(b)
F : Acetaminophen model of acute liver injury and failure. (a) Liver confocal intravital microscopy showing that while control mice
have a well perfused liver vasculature (evidenced by PE-anti-CD antibody staining, in red) and absence of necrosis (in green), APAP-
treated mice ( mg/Kg) had several dark areas in the red channel, which are suggestive of malperfusion caused by necrosis (elicited by
DNA staining using Sytox Green). (b) Serum liver transaminase levels conrmed necrosis. Statistical signicance in comparison to controls
(𝑃 < 0.05). A x objective was used during imaging. One-way ANOVA with Tukey’s post hoc test. Data are expressed as mean ±S.E.M(𝑛=6
mice/treatment).
APAP
Veh i c l e
(a)
0
1
2
3
Histological score
Veh i c l e A PAP
0
100
200
300
400
Neutrophils (×104)
Veh i c l e A PAP
0
20
40
60
80
Necrosis area (%)
Veh i c l e A PAP
(b)
F : Histopathological assessment of acetaminophen-induced liver injury (APAP). (a) Liver histology (H&E) showing that while control
mice have a healthy liver architecture and absence of necrosis, APAP-treated mice had several necrotic areas with leukocyte inltration (green
arrow). (b) Digital quantication of necrotic areas using ImageJ, histopathological score, and neutrophil inltration (assessed by MPO levels)
conrmed severe liver inammation and necrosis induced by cocaine. Statistical signicance in comparison to controls (𝑃 < 0.05). One-way
ANOVA with Tukey’s post hoc test. Data are expressed as mean ±S.E.M(𝑛=6mice/treatment).
Mediators of Inammation
Veh APAP 600 APAP 800
0
10
20
30
40
50
60
70
80
90
100
5/9
1/12
3/4
Lethality (%)
P = 0.04
P = 0.02
Cocaine (75 mg/kg)
(a)
Veh APAP 600 APAP 800
∗∗∗
0
10
20
30
40
50
60
70
80
90
100
Seizures (%)
P < 0.0001
Cocaine (75 mg/kg)
(b)
Veh APAP 600 APAP 800
0
10
20
30
40
50
∗∗∗
Seizure duration (s)
Cocaine (75 mg/kg)
(c)
Veh APAP 600 APAP 800
0
100
200
300
400
500
600 ∗∗
Latency (s)
Cocaine (75 mg/kg)
(d)
F : Previous hepatic injury ( hours) with APAP interferes in seizure dynamics. (a) Administration of APAP at doses of 
and  mg/kg  hours before cocaine administration increased lethality when compared to group vehicle/cocaine. (b) APAP at dose of
 mg/kg reduced percentage of seizures and its duration (c). (d) Previous administration of APAP at dose of  mg/kg decreased seizure
latency.APAP at dose of  mg/kg enhanced seizure latency. Signicantly dierent from vehicle group (𝑃< 0.05). ∗∗Signicantly dierent
from vehicle group (∗∗𝑃< 0.001). ∗∗∗Signicantly dierent from vehicle group (∗∗∗𝑃< 0.0001). One-way ANOVA with Tukey’s post hoc
test. Data are expressed as mean ±S.E.M(𝑛=6mice/treatment).
besides being well-tolerated and exhibiting a broad spec-
trum of therapeutic properties []. us, it is currently
attracting considerable interest as a potential medicine due
to its anti-inammatory, neuroprotective, antipsychotic, anx-
iolytic, antiepileptic, and anticancer eects [,,].
OneofthemainactionsofCBDisitsanticonvulsant
eect. In many experimental models, it exerted antiseizure
activities against PTZ-induced generalized seizures in rats
[], in the pilocarpine model temporal lobe seizures and the
penicillin model of partial seizures []aswellasinthePTZ-
and electroshock-induced seizures in rats []. us, consid-
eringtheecacyofCBDandthefactthatitdoesnotshare
thepsychotomimetic,amnestic,andsedativeeectsofΔ9-
THC, it is an interesting potential treatment for epilepsies. A
recently clinical study observed that CBD-enriched cannabis
extracts signicantly reduced seizure frequency in children
with treatment-resistant epilepsy, indicating that it could be
used as a strategy when other pharmacological treatments fail
[].
In addition, the mechanisms of action of this compound
have remained elusive and should be further investigated.
CBD is a compound with a wide plethora of pharmacolog-
ical actions, including anti-inammatory and antioxidative
eects, which may act through diverse mechanisms, facilitat-
ing the endocannabinoid system by blocker FAAH enzyme,
binding to TRPV channels, serotonin type-A receptors,
AA adenosine receptors, and peroxisome proliferator-
activated receptors (i.e., PPAR-𝛾)amongothers[,,].
In most in vivo models of inammation, CBD attenuates
inammatory cell migration/inltration (e.g., neutrophils)
[]. CBD suppresses T cell responses, decreases the release
of bioactive tumour necrosis factor (TNF𝛼), and reduces
prostaglandin E (PGE) levels, cyclooxygenase (COX) activ-
ity, and production of nitric oxide (NO). e suppressive
eects of CBD on cellular immune responses and on the
production of proinammatory mediators may indicate its
usefulness in several inammatory diseases []. Addition-
ally,CBDadministrationwasabletodecreaseleukocyte
migration into the lungs, myeloperoxidase activity in the lung
tissue, protein concentration and production of proinam-
matory cytokines (TNF𝛼and IL-), and chemokines (MCP-
 and MIP-) in the bronchoalveolar lavage supernatant
showing a potent anti-inammatory eect in mice submitted
to LPS-induced acute lung injury []. In other context,
CBD modied the deleterious eects of inammation in
a viral model of multiple sclerosis through decreasing the
transmigration of blood leukocytes by downregulating the
expression of vascular cell adhesion molecule- (VCAM-),
chemokines (CCL and CCL), and the proinammatory
cytokine IL-𝛽[].
Mediators of Inammation
DNA (necrosis) Merge
PECAM-1
(sinusoids)
Cocaine
CBD ×cocaine
(a)
0
1000
2000
3000
ALT (U/L)
Cocaine
Control Veh C B D URB
∗∗
(b)
**
Cocaine
Control Veh CBD URB
0
10
20
30
40
50
Cardiogreen (𝜇g/mL)
∗∗
(c)
F : Cannabidiol (CBD;  mg/Kg) prevented cocaine-induced liver injury. (a) Liver confocal intravital microscopy showing that while
CBD-treated mice have a well perfused liver vasculature (evidenced by PE-anti-CD antibody staining, in red) and absence of necrosis (in
green), cocaine-treated mice ( mg/Kg) had several dark areas in the red channel, which are suggestive of malperfusion caused by necrosis
(elicited by DNA staining using Sytox Green). (b) Reduced serum liver transaminase levels and (c) clearance of cardiogreen conrmed
inhibition of cocaine-induced liver injury by CBD. However, inhibition of anandamide hydrolysis by URB had no detectable eect. Statistical
signicance in comparison to controls (𝑃 < 0.05)and ∗∗in comparison to vehicle-treated group. A x objective was used during imaging.
One-way ANOVA with Tukey’s post hoc test. Data are expressed as mean ±S.E.M(𝑛=6mice/treatment).
In respect to eects of CBD on liver, recent data showed
that CBD protected against liver toxicity induced by a single
dose of cadmium chloride []. Similarly, CBD attenuated
the deterioration in the measured biochemical parameters
and damages mediated by ischemia/reperfusion liver injury
besides reducing the inammatory response in tissue liver
[]. Previous studies already revealed that CBD pretreat-
ment signicantly protected against liver ischemia for  min
followed by reperfusion for  hs [].
Finally, our results on peripheric and central eects of
CBD against toxic eects of cocaine are in agreement with
previous results showing that this compound restored both
liver and brain function in a model of hepatic encephalopathy
associated with fulminant hepatic failure induced in mice by
thioacetamide []. Pretreatment with CBD was able to revert
neurological and cognitive deterioration induced by thioac-
etamideinparallelwithincreasesofliverfunction,conrmed
by improve of ammonia, bilirubin, aspartate transaminase
(AST), and alanine transaminase (ALT) levels. ese results
showed that CBD eects might result from a combination of
its actions in the liver and brain.
In conclusion, the present study demonstrates that co-
caine-induced seizures occur along with severe hepatic tox-
icity. e phytocannabinoid, CBD, but not the anandamide
 Mediators of Inammation
Cocaine
Veh i c l e CBD ×cocaine
(a)
0
20
40
60
Cocaine
Necrosis area (%)
Veh CBD URBControl
∗∗ 0
1
2
3
Cocaine
Histological score
Veh CBD URBControl
∗∗
0
50
100
150
200
Cocaine
Neutrophils (×104)
Veh CBD URBControl
∗∗
(b)
Veh CBD URB
0
100
200
300
400
500
600
Cocaine
Seizure latency (s)
∗∗ ∗∗
Veh CBD URB
0
10
20
30
40
50
Cocaine
Seizure duration (s)
∗∗ ∗∗
(c)
F : Cannabidiol (CBD;  mg/Kg) prevented liver inammation and seizure due to cocaine intake. (a) Liver histology showing that
while CBD-treated mice have a normal liver architecture, cocaine-treated mice ( mg/Kg) had extensive areas which were suggestive of
malperfusion caused by necrosis. (b) Reduced necrotic area, histological score, and neutrophil inltration conrmed the anti-inammatory
and protective eects of CBD duringcocaine-induced hepatotoxicity. However, in hibition of anandamide hydrolysis by URB had no detectable
eect on liver parameters. (c) Despite lack of eect on liver injury, both CBD and URB inhibited cocaine-induced seizure, increasing latency
and reducing seizure duration. Statistical signicance in comparison to controls (𝑃 < 0.05)and ∗∗in comparison to vehicle-treated group.
A x objective was used during imaging. One-way ANOVA with Tukey’s post hoc test. Data are expressed as mean ±S.E.M(𝑛=6
mice/treatment).
hydrolysis inhibitor, URB, protects against both eects
and warrants further investigation as a potential treatment for
both brain and liver consequences of cocaine intoxication.
Conflict of Interests
e authors declare that none of the authors have conict of
interests.
Authors’ Contribution
Luciano Rezende Vilela and Lindisley Ferreira Gomides
contributed equally to this work.
Acknowledgments
is work was supported by CNPq, CAPES, and Fapemig.
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... The hepatoprotective and neuroprotective effects of CBD obtained in this study are also supported by data from the literature indicating other protective mechanisms beyond antioxidant activity. Studies have shown that CBD may have a protective effect on the liver and brain from the toxic effects of cocaine or alcohol, minimizing inflammatory damage [40,43]. CBD reduces alcohol-induced liver damage by decreasing lipid accumulation and oxidative stress, stimulating autophagy and modulating the inflammatory response. ...
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The growing awareness and need to protect public health, including food safety, require a thorough study of the mechanism of action of veterinary drugs in consumers to reduce their negative impact on humans. Inappropriate use of veterinary drugs in animal husbandry, such as tiamulin, leads to the appearance of residues in edible animal tissues. The use of natural substances of plant origin, extracted from hemp (Cannabis sativa L.), such as cannabidiol (CBD), is one of the solutions to minimize the negative effects of tiamulin. This study aimed to determine the effect of CBD on the cytotoxicity of tiamulin in humans. The cytotoxic activity of tiamulin and the effect of its mixtures with CBD were tested after 72 h exposure to three human cell lines: SH-SY5Y, HepG2 and HEK-293. Cytotoxic concentrations (IC50) of the tested drug and in combination with CBD were assessed using five biochemical endpoints: mitochondrial and lysosomal activity, proliferation, cell membrane integrity and effects on DNA synthesis. Oxidative stress, cell death and cellular morphology were also assessed. The nature of the interaction between the veterinary drug and CBD was assessed using the combination index. The long-term effect of tiamulin inhibited lysosomal (SH-SY5SY) and mitochondrial (HepG2) activity and DNA synthesis (HEK-293). IC50 values for tiamulin ranged from 2.1 to >200 µg/mL (SH-SY5SY), 13.9 to 39.5 µg/mL (HepG2) and 8.5 to 76.9 µg/mL (HEK-293). IC50 values for the drug/CBD mixtures were higher. Reduced levels of oxidative stress, apoptosis and changes in cell morphology were demonstrated after exposure to the mixtures. Interactions between the veterinary drug and CBD showed a concentration-dependent nature of tiamulin in cell culture, ranging from antagonistic (low concentrations) to synergistic effects at high drug concentrations. The increased risk to human health associated with the presence of the veterinary drug in food products and the protective nature of CBD use underline the importance of these studies in food toxicology and require further investigation.
... The consumption of cannabidiol (CBD) products in the United States and European societies has been on the rise for several years (Cannaccord Genuity 2019; European Drug Report 2023). Athletes also regularly use CBD products in the belief to promote their regeneration and increase their performance (Brisola-Santos et al. 2016;Zeiger et al. 2019;Ware et al. 2018;Vilela et al. 2015;Docter et al. 2020). Initial results suggest that CBD has a slight inhibitory effect on muscle damage (Isenmann et al. 2024;Isenmann et al. 2021;Veit et al. 2020), but does not influence muscle soreness (Alpy et al. 2023). ...
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The side effects and safety of cannabidiol (CBD) products are currently discussed in different contexts. Of all adverse effects, hepatotoxic effects have been reported most frequently in previous studies. However, the threshold for liver toxicity of CBD in humans is uncertain due to the lack of adequately designed studies in humans below the lowest observed adverse effect level (LOAEL) of 300 mg/day. In a randomised, three-arm, double-blind, crossover study, the effects of two CBD products (oil and solubilisate (solu) containing 60 mg CBD) were investigated during a high-intensity exercise protocol. Seventeen well-trained subjects (26±4 years, 181±5 cm, 85.6±9.4 kg) participated in the intervention. All subjects were healthy and had no physiological or psychological injuries. Participants were divided into advanced (Ad) and highly advanced (Hi) athletes … They consumed 60 mg of the compound in each microcycle over 7 days. To evaluate possible effects of short-term repeated use of 60 mg CBD on oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT), gamma-glutamyl transferase (GGT) and creatinine (CREA) were analysed before and after each microcycle. GOT increased significantly in both performance levels of the placebo groups (Ad: p≤0.001; Hi: p=0.003). This increase was significantly reduced in the Ad group by both CBD oil (p=0.050, ES=0.66) and CBD solu (p=0.027; ES=0.75). GPT also increased significantly in both placebo groups (Ad: p≤0.001; Hi: p=0.032). This increase was significantly reduced in the Ad group by both CBD oil (p=0.027; ES=0.75) and CBD solu (p=0.023; ES=0.77). These effects were not observed in the Hi group for either parameter. Our results show that short-term repeated use of 60 mg CBD can inhibit exercise-induced liver activity. Furthermore, under the conditions of the present study, there was no evidence for hepatotoxic effects of oral intake of CBD at 60 mg for seven days. Nevertheless, despite the inhibitory effect on exercise-induced liver activity, the study provides evidence for the pharmacological effects of CBD on the liver even at low CBD dose and does not exclude adverse effects in sensitive individuals.
... Patra et al. (2020) recently reported that CBD prevented premature mortality and improved cognition and social interaction in the Scn1a ± mouse model of Dravet syndrome. Furthermore, CBD increased the mean survival time in aluminium phosphide-poisoned rats (Hooshangi Shayesteh et al., 2022) and reduced cocaine lethality in mice (Vilela et al., 2015). Altogether, these findings indicate that CBD might exert a neuroprotective effect by decreasing mortality in rats under different experimental conditions. ...
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Objective To investigate the effects of cannabidiol (CBD) on emotional and cognitive symptoms in rats with intra-nigral 6-hydroxydopamine (6-OHDA) lesions. Methods Adult male Wistar rats received bilateral intranigral 6-OHDA infusions and were tested in a battery of behavioral paradigms to evaluate nonmotor symptoms. The brains were obtained to evaluate the effects of CBD on hippocampal neurogenesis. Results 6-hydroxydopamine-lesioned rats exhibited memory impairments and despair-like behavior in the novelty-suppressed feeding test and forced swim test, respectively. The animals also exhibited dopaminergic neuronal loss in the substantia nigra pars compacta (SNpc), striatum, and ventral tegmental area and a reduction of hippocampal neurogenesis. Cannabidiol decreased dopaminergic neuronal loss in the SNpc, reduced the mortality rate and decreased neuroinflammation in 6-OHDA-lesioned rats. In parallel, CBD prevented memory impairments and attenuated despair-like behavior that were induced by bilateral intranigral 6-OHDA lesions. Repeated treatment with CBD favored the neuronal maturation of newborn neurons in the hippocampus in Parkinsonian rats. Conclusion The present findings suggest a potential beneficial effect of CBD on nonmotor symptoms induced by intra-nigral 6-OHDA infusion in rats.
... CBD safeguards the liver from the toxic effects of cadmium by inhibiting hepatic lipid peroxidation, preventing the depletion of reduced glutathione and nitric oxide, and inhibiting hydrogen peroxide enzyme activity [55]. Also, CBD alleviated acute liver inflammation and injury induced by cocaine and prevented seizures caused by cocaine toxicity [56]. CBD-rich mustard extracts can inhibit oxidative stress, gluconeogenesis, and hepatic lipid accumulation in ironmediated oxidative liver damage while regulating cholinergic and purinergic activities [57]. ...
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Cannabidiol (CBD), a non-psychoactive phytocannabinoid abundant in Cannabis sativa, has gained considerable attention for its anti-inflammatory, antioxidant, analgesic, and neuroprotective properties. It exhibits the potential to prevent or slow the progression of various diseases, ranging from malignant tumors and viral infections to neurodegenerative disorders and ischemic diseases. Metabolic dysfunction-associated steatotic liver disease (MASLD), formerly known as non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease, and viral hepatitis stand as prominent causes of morbidity and mortality in chronic liver diseases globally. The literature has substantiated CBD’s potential therapeutic effects across diverse liver diseases in in vivo and in vitro models. However, the precise mechanism of action remains elusive, and an absence of evidence hinders its translation into clinical practice. This comprehensive review emphasizes the wealth of data linking CBD to liver diseases. Importantly, we delve into a detailed discussion of the receptors through which CBD might exert its effects, including cannabinoid receptors, CB1 and CB2, peroxisome proliferator-activated receptors (PPARs), G protein-coupled receptor 55 (GPR55), transient receptor potential channels (TRPs), and their intricate connections with liver diseases. In conclusion, we address new questions that warrant further investigation in this evolving field.
... This study indicates that 4-F-MDMB-BUTINACA has high toxicity, its toxic effects on liver tissue were close to the effect of cocaine, which is classified as highly toxic and causes liver damage and inflammation. 25 The inflammation, necrosis, and congestion seen in kidneys of 4-F-MDMB-BUTINACA -treated mice were similar to the changes found by Dargan et al. (2014), who studied the effects of methoxetamine (analogue of ketamine) in renal toxicity in mice. 26 Conclusion 4-F-MDMB-BUTINACA is of high acute toxicity but had rapid effects after administration. ...
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The purpose of this research was to evaluate the single-dose toxicity of 4-F-MDMB-BUTINACA in Swiss albinomice using histopathological analysis of liver and kidney specimens.The experimental protocol included oral treatment of mice with different doses (5, 50, 300, 2000 mg/kg bodyweight of 4-F-MDMB-BUTINACA) for 24 hours. At the end of the treatment, blood samples had been drawn, andrenal and hepatic tissues have been excised from the experimental mice groups for histological examinations.The results revealed that dose-dependent treatment with 4-F-MDMB-BUTINACA causes mild tremor clinicalsigns with low doses and photophobia (sensitivity to light) and even cessation of breathing as a potential cause ofdeath with high doses in treated mice.The LD50 value of 4-F-MDMB-BUTINACA was 32.60 mg/kg, which is considered as a chemical compound of lowtoxicity. Histological studies confirmed that liver and kidney toxicities have been manifested in the findings ofcongestion, necrosis, inflammation, and bleeding within the liver and to lesser extent in the kidneys.
... Notably, these protective effects are independent of CB1R/CB2R [128,132]. Other studies show that CBD has beneficial effects on the liver and brain [133,134]. ...
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Background The NLRP3 inflammasome is a vital player in the emergence of inflammation. The priming and activation of the NLRP3 inflammasome is a major trigger for inflammation which is a defense response against adverse stimuli. However, the excessive activation of the NLRP3 inflammasome can lead to the development of various inflammatory diseases. Cannabidiol, as the second-most abundant component in cannabis, has a variety of pharmacological properties, particularly anti-inflammation. Unlike tetrahydrocannabinol, cannabidiol has a lower affinity for cannabinoid receptors, which may be the reason why it is not psychoactive. Notably, the mechanism by which cannabidiol exerts its anti-inflammatory effect is still unclear. Methods We have performed a literature review based on published original and review articles encompassing the NLRP3 inflammasome and cannabidiol in inflammation from central databases, including PubMed and Web of Science. Results and conclusions In this review, we first summarize the composition and activation process of the NLRP3 inflammasome. Then, we list possible molecular mechanisms of action of cannabidiol. Next, we explain the role of the NLRP3 inflammasome and the anti-inflammatory effect of cannabidiol in inflammatory disorders. Finally, we emphasize the capacity of cannabidiol to suppress inflammation by blocking the NLRP3 signaling pathway, which indicates that cannabidiol is a quite promising anti-inflammatory compound.
... Entre as doenças mais tratadas, de acordo com os conhecimentos dos estudantes, os distúrbios relacionados ao sistema nervoso foram os mais citados, sinalizando que os mesmos detêm algum conhecimento sobre a efetividade da Cannabis. Pesquisas demonstram que a Cannabis tem apresentado resultados positivos no tratamento de distúrbios neurológicos, tais como a depressão, ansiedade, epilepsia (LUVONE et al., 2009;VILELA et al., 2015). Por exemplo, pesquisas de Devinsky et al., (2016) e Rosenberg et al., (2017, compararam a eficácia de medicamentos extraídos da Cannabis com a de antiepilépticos convencionais comercializados, demonstraram a redução em quase 50% na frequência de crises por mês. ...
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Cannabis was used to treat convulsions and other disorders since ancient times. In the last few decades, preclinical animal studies and clinical investigations have established the role of cannabidiol (CBD) in treating epilepsy and seizures and support potential therapeutic benefits for cannabinoids in other neurological and psychiatric disorders. Here, we comprehensively review the role of cannabinoids in epilepsy. We briefly review the diverse physiological processes mediating the central nervous system response to cannabinoids, including D ⁹ -THC, cannabidiol, and terpenes. Next, we characterize the anti- and proconvulsive effects of cannabinoids from animal studies of acute seizures and chronic epileptogenesis. We then review the clinical literature on using cannabinoids to treat epilepsy, including anecdotal evidence and case studies as well as the more recent randomized-controlled clinical trials that led to FDA approval of CBD for some types of epilepsy. Overall, we seek to evaluate our current understanding of cannabinoids in epilepsy and focus future research on unanswered questions.
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Background: Preclinical and clinical data suggest that cannabidiol (CBD), a major non-psychotomimetic compound from Cannabis sativa, induces antipsychotic-like effects. However, the antipsychotic properties of repeated CBD treatment have been poorly investigated. Behavioral changes induced by repeated treatment with glutamate N-methyl-D-aspartate receptor (NMDAR) antagonists have been proposed as an animal model of schizophrenia-like signs. In the present study, we evaluated if repeated treatment with CBD would attenuate the behavioral and molecular modifications induced by chronic administration of one of these antagonists, MK-801. Methods: Male C57BL/6J mice received daily i.p. injections of MK-801 (0.1, 0.5, or 1mg/kg) for 14, 21, or 28 days. Twenty-four hours after the last injection, animals were submitted to the prepulse inhibition (PPI) test. After that, we investigated if repeated treatment with CBD (15, 30, and 60mg/kg) would attenuate the PPI impairment induced by chronic treatment with MK-801 (1mg/kg; 28 days). CBD treatment began on the 6th day after the start of MK-801 administration and continued until the end of the treatment. Immediately after the PPI, the mice brains were removed and processed to evaluate the molecular changes. We measured changes in FosB/ΔFosB and parvalbumin (PV) expression, a marker of neuronal activity and a calcium-binding protein expressed in a subclass of GABAergic interneurons, respectively. Changes in mRNA expression of the NMDAR GluN1 subunit gene (GRN1) were also evaluated. CBD effects were compared to those induced by the atypical antipsychotic clozapine. Results: MK-801 administration at the dose of 1mg/kg for 28 days impaired PPI responses. Chronic treatment with CBD (30 and 60mg/kg) attenuated PPI impairment. MK-801 treatment increased FosB/ΔFosB expression and decreased PV expression in the medial prefrontal cortex. A decreased mRNA level of GRN1 in the hippocampus was also observed. All the molecular changes were attenuated by CBD. CBD by itself did not induce any effect. Moreover, CBD effects were similar to those induced by repeated clozapine treatment. Conclusions: These results indicate that repeated treatment with CBD, similar to clozapine, reverses the psychotomimetic-like effects and attenuates molecular changes observed after chronic administration of an NMDAR antagonist. These data support the view that CBD may have antipsychotic properties.
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Abstract We have previously shown that the prophylactic treatment with cannabidiol (CBD) reduces inflammation in a model of acute lung injury (ALI). In this work we analyzed the effects of the therapeutic treatment with CBD in mice subjected to the model of lipopolysaccharide (LPS)-induced ALI on pulmonary mechanics and inflammation. CBD (20 and 80 mg/kg) was administered (i.p.) to mice 6 h after LPS-induced lung inflammation. One day (24 h) after the induction of inflammation the assessment of pulmonary mechanics and inflammation were analyzed. The results show that CBD decreased total lung resistance and elastance, leukocyte migration into the lungs, myeloperoxidase activity in the lung tissue, protein concentration and production of pro-inflammatory cytokines (TNF and IL-6) and chemokines (MCP-1 and MIP-2) in the bronchoalveolar lavage supernatant. Thus, we conclude that CBD administered therapeutically, i.e. during an ongoing inflammatory process, has a potent anti-inflammatory effect and also improves the lung function in mice submitted to LPS-induced ALI. Therefore the present and previous data suggest that in the future cannabidiol might become a useful therapeutic tool for the attenuation and treatment of inflammatory lung diseases.
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Isolation and structure elucidation of most of the major cannabinoid constituents - including Δ(9)-tetrahydrocannabinol (Δ(9)-THC), which is the principal psychoactive molecule in Cannabis sativa - was achieved in the 1960s and 1970s. It was followed by the identification of two cannabinoid receptors in the 1980s and the early 1990s and by the identification of the endocannabinoids shortly thereafter. There have since been considerable advances in our understanding of the endocannabinoid system and its function in the brain, which reveal potential therapeutic targets for a wide range of brain disorders.
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Unlabelled: Drug-induced liver injury (DILI) is an important cause of acute liver failure, with limited therapeutic options. During DILI, oncotic necrosis with concomitant release and recognition of intracellular content amplifies liver inflammation and injury. Among these molecules, self-DNA has been widely shown to trigger inflammatory and autoimmune diseases; however, whether DNA released from damaged hepatocytes accumulates into necrotic liver and the impact of its recognition by the immune system remains elusive. Here we show that treatment with two different hepatotoxic compounds (acetaminophen and thioacetamide) caused DNA release into the hepatocyte cytoplasm, which occurred in parallel with cell death in vitro. Administration of these compounds in vivo caused massive DNA deposition within liver necrotic areas, together with an intravascular DNA coating. Using confocal intravital microscopy, we revealed that liver injury due to acetaminophen overdose led to a directional migration of neutrophils to DNA-rich areas, where they exhibit an active patrolling behavior. DNA removal by intravenous DNASE1 injection or ablation of Toll-like receptor 9 (TLR9)-mediated sensing significantly reduced systemic inflammation, liver neutrophil recruitment, and hepatotoxicity. Analysis of liver leukocytes by flow cytometry revealed that emigrated neutrophils up-regulated TLR9 expression during acetaminophen-mediated necrosis, and these cells sensed and reacted to extracellular DNA by activating the TLR9/NF-κB pathway. Likewise, adoptive transfer of wild-type neutrophils to TLR9(-/-) mice reversed the hepatoprotective phenotype otherwise observed in TLR9 absence. Conclusion: Hepatic DNA accumulation is a novel feature of DILI pathogenesis. Blockage of DNA recognition by the innate immune system may constitute a promising therapeutic venue.
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To propose an alternative model of hepatic encephalopathy (HE) in mice, resembling the human features of the disease. Mice received two consecutive intraperitoneal injections of thioacetamide (TAA) at low dosage (300 mg/kg). Liver injury was assessed by serum transaminase levels (ALT) and liver histology (hematoxylin and eosin). Neutrophil infiltration was estimated by confocal liver intravital microscopy. Coagulopathy was evaluated using prolonged prothrombin and partial thromboplastin time. Hemodynamic parameters were measured through tail cuff. Ammonia levels were quantified in serum and brain samples. Electroencephalography (EEG) and psychomotor activity score were performed to show brain function. Brain edema was evaluated using magnetic resonance imaging. Mice submitted to the TAA regime developed massive liver injury, as shown by elevation of serum ALT levels and a high degree of liver necrosis. An intense hepatic neutrophil accumulation occurred in response to TAA-induced liver injury. This led to mice mortality and weight loss, which was associated with severe coagulopathy. Furthermore, TAA-treated mice presented with increased serum and cerebral levels of ammonia, in parallel with alterations in EEG spectrum and discrete brain edema, as shown by magnetic resonance imaging. In agreement with this, neuropsychomotor abnormalities ensued 36 h after TAA, fulfilling several HE features observed in humans. In this context of liver injury and neurological dysfunction, we observed lung inflammation and alterations in blood pressure and heart rate that were indicative of multiple organ dysfunction syndrome. In summary, we describe a new murine model of hepatic encephalopathy comprising multiple features of the disease in humans, which may provide new insights for treatment.
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Delta(9)-tetrahydrocannabinol binds cannabinoid (CB(1) and CB(2)) receptors, which are activated by endogenous compounds (endocannabinoids) and are involved in a wide range of physiopathological processes (e.g. modulation of neurotransmitter release, regulation of pain perception, and of cardiovascular, gastrointestinal and liver functions). The well-known psychotropic effects of Delta(9)-tetra hydrocannabinol, which are mediated by activation of brain CB(1) receptors, have greatly limited its clinical use. However, the plant Cannabis contains many cannabinoids with weak or no psychoactivity that, therapeutically, might be more promising than Delta(9)-tetra hydrocannabinol. Here, we provide an overview of the recent pharmacological advances, novel mechanisms of action, and potential therapeutic applications of such non-psychotropic plant-derived cannabinoids. Special emphasis is given to cannabidiol, the possible applications of which have recently emerged in inflammation, diabetes, cancer, affective and neurodegenerative diseases, and to Delta(9)-tetrahydrocannabivarin, a novel CB(1) antagonist which exerts potentially useful actions in the treatment of epilepsy and obesity.
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The treatment for cocaine use constitutes a clinical challenge because of the lack of appropriate therapies and the high rate of relapse. Recent evidence indicates that the immune system might be involved in the pathogenesis of cocaine addiction and its co-morbid psychiatric disorders. This work examined the plasma pro-inflammatory cytokine and chemokine profile in abstinent cocaine users (n = 82) who sought outpatient cocaine treatment and age/sex/body mass-matched controls (n = 65). Participants were assessed with the diagnostic interview Psychiatric Research Interview for Substance and Mental Diseases according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision (DSM-IV-TR). Tumor necrosis factor-alpha, chemokine (C-C motif) ligand 2/monocyte chemotactic protein-1 and chemokine (C-X-C motif) ligand 12 (CXCL12)/stromal cell-derived factor-1 (SDF-1) were decreased in cocaine users, although all cytokines were identified as predictors of a lifetime pathological use of cocaine. Interleukin-1 beta (IL-1β), chemokine (C-X3 -C motif) ligand 1 (CX3CL1)/fractalkine and CXCL12/SDF-1 positively correlated with the cocaine symptom severity when using the DSM-IV-TR criteria for cocaine abuse/dependence. These cytokines allowed the categorization of the outpatients into subgroups according to severity, identifying a subgroup of severe cocaine users (9-11 criteria) with increased prevalence of co-morbid psychiatric disorders [mood (54%), anxiety (32%), psychotic (30%) and personality (60%) disorders]. IL-1β was observed to be increased in users with such psychiatric disorders relative to those users with no diagnosis. In addition to these clinical data, studies in mice demonstrated that plasma IL-1β, CX3CL1 and CXCL12 were also affected after acute and chronic cocaine administration, providing a preclinical model for further research. In conclusion, cocaine exposure modifies the circulating levels of pro-inflammatory mediators. Plasma cytokine/chemokine monitoring could improve the stratification of cocaine consumers seeking treatment and thus facilitate the application of appropriate interventions, including management of heightened risk of psychiatric co-morbidity. Further research is necessary to elucidate the role of the immune system in the etiology of cocaine addiction.
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To present a summary of current scientific evidence about the cannabinoid, cannabidiol (CBD) with regard to its relevance to epilepsy and other selected neuropsychiatric disorders. We summarize the presentations from a conference in which invited participants reviewed relevant aspects of the physiology, mechanisms of action, pharmacology, and data from studies with animal models and human subjects. Cannabis has been used to treat disease since ancient times. Δ9-Tetrahydrocannabinol (Δ9-THC) is the major psychoactive ingredient and CBD is the major nonpsychoactive ingredient in cannabis. Cannabis and Δ9-THC are anticonvulsant in most animal models but can be proconvulsant in some healthy animals. The psychotropic effects of Δ9-THC limit tolerability. CBD is anticonvulsant in many acute animal models, but there are limited data in chronic models. The antiepileptic mechanisms of CBD are not known, but may include effects on the equilibrative nucleoside transporter; the orphan G-protein-coupled receptor GPR55; the transient receptor potential of vanilloid type-1 channel; the 5-HT1a receptor; and the α3 and α1 glycine receptors. CBD has neuroprotective and antiinflammatory effects, and it appears to be well tolerated in humans, but small and methodologically limited studies of CBD in human epilepsy have been inconclusive. More recent anecdotal reports of high-ratio CBD:Δ9-THC medical marijuana have claimed efficacy, but studies were not controlled. CBD bears investigation in epilepsy and other neuropsychiatric disorders, including anxiety, schizophrenia, addiction, and neonatal hypoxic-ischemic encephalopathy. However, we lack data from well-powered double-blind randomized, controlled studies on the efficacy of pure CBD for any disorder. Initial dose-tolerability and double-blind randomized, controlled studies focusing on target intractable epilepsy populations such as patients with Dravet and Lennox-Gastaut syndromes are being planned. Trials in other treatment-resistant epilepsies may also be warranted.A PowerPoint slide summarizing this article is available for download in the Supporting Information section here.