European Journal of Pharmacology 702 (2013) 2531

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European Journal of Pharmacology

journal homepage: www.elsevier.com/locate/ejphar

Behavioural pharmacology

Effects of dopamine receptor agonist and antagonists

on cholestasis-induced anxiolytic-like behaviors in rats
Mohammad Reza Zarrindast a,b,c,d,e,n, Delaram Eslimi Esfahani f, Shahrbano Oryan f,
Mohammad Nasehi g, Mohammad Torabi Nami e,h
a
Department of Neuroscience, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran, Iran
b
Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
c
Iranian National Centre for Addiction Studies, Tehran University of Medical Sciences, Tehran, Iran
d
School of Cognitive Sciences, Institute for Research in Fundamental Sciences (IPM), Tehran, Iran
e
Institute for Cognitive Science Studies (ICSS), Tehran, Iran
f
Faculty of Biological Sciences, Kharazmi (Tarbiat Moalem) University, Tehran, Iran
g
Department of Biology, Faculty of Basic Sciences, Islamic Azad University, Garmsar branch, Semnan, Iran
h
Department of Neuroscience, School of Advanced Medical Science and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran

a r t i c l e i n f o abstract

Article history: Dysfunctions in the dopamine transmission system have been suggested to contribute to the
Received 17 April 2012 pathogenesis of hepatic encephalopathy. In an experimental animal model, cholestasis induction
Received in revised form through bile duct ligation may present several main pathological features of hepatic encephalopathy.
20 December 2012
Dopaminergic systems are shown to play pivotal roles in regulation of anxiety-like behaviors. The main
Accepted 15 January 2013
Available online 29 January 2013
bile duct in male Wistar rats, weighing 220240 g, was ligated using two ligatures plus duct transection
in between. Anxiety-like behaviors were measured using the elevated plus maze task. Cholestasis
Keywords: increased the open arm time percentage (%OAT), 13 but not 10 days after bile duct ligation, indicating
Cholestasis an anxiolytic-like effect. Sole intraperitoneal injection of apomorphine (dopamine D1/D2 receptor
Apomorphine
agonist, 0.25 mg/kg), SCH23390 (dopamine D1 receptor antagonist, 0.005, 0.01 and 0.02 mg/kg) or
SCH23390
sulpiride (dopamine D2 receptor antagonist, 0.125, 0.25 and 0.5 mg/kg) did not alter %OAT, open arm
Sulpiride
Anxiety entries percentage (%OAE) and locomotor activity in the sham-operated rats. Meanwhile, the higher
Elevated plus maze dose apomorphine (0.5 mg/kg) induced anxiolytic-like behaviors in this group. The subthreshold dose
Rat(s) injection of SCH23390 or sulpiride, partially reversed the anxiolytic-like behaviors induced by
cholestasis (13 days after bile duct ligation). On the other hand, subthreshold dose of apomorphine
in cholestatic rats (10 days post bile duct ligation) induced anxiolytic-like effects which could be
blocked by SCH23390 or sulpiride. The effective doses of above drugs did not alter locomotor activity,
number of rearings, groomings and defections. These ndings suggested that the dopaminergic system
may potentially be involved in the modulation of cholestasis-induced anxiolytic-like behaviors in rats.
& 2013 Elsevier B.V. All rights reserved.

1. Introduction have revealed that cholestasis decreases the anxiety-like beha-

Acute and chronic failure in liver function may give rise to
cognitive and non-cognitive impairments in the brain, namely;
hepatic encephalopathy (Huang et al., 2010; Magen et al., 2010;
Zarrindast et al., 2012a). Cholestasis which leads to hepatic
encephalopathy (Garcia-Moreno et al., 2005) is potentially asso-
ciated with liver cirrhosis (Ferenci et al., 2002). The most
frequently applied experimental model for induction of choles-
tasis is common bile duct ligation (Pi-Chieh Wang et al., 2011;
Zarrindast et al., 2012a; Zhang et al., 2012). Some investigations
n
Corresponding author at: Department of Neuroscience, School of Advanced
Medical Technologies, Tehran University of Medical Sciences, P.O. Box 13145-784,
Tehran, Iran. Tel./fax: 98 21 66402569.
E-mail address: zarinmr@ams.ac.ir (M. Reza Zarrindast).
viors (Eslimi et al., 2011), induces memory decits (Cauli et al.,
2009; Huang et al., 2009; Magen et al., 2010; Zarrindast et al.,
2012b), tremor (Chung et al., 2005) and alters the sleep pattern
(Newton, 2008). It has been made clear that cholestasis alters the
activity of all classic neurotransmitter systems such as opioidergic
(Roberts et al., 1987; Zhang et al., 2004) and dopaminergic (Glaser
et al., 2003, 2006; Zimatkin et al., 2008).
Several studies have also indicated that the two major dopa-
minergic systems including mesolimbic and mesocortical are
involved in anxiogenic- and anxiolytic-like responses induced
by some medications or acute stress (Cabib et al., 1988; Deutch
et al., 1985; Dunn, 1988; Feenstra et al., 1995; Imperato et al.,
1990; Louilot et al., 1986; Nasehi et al., 2011; Puglisi-Allegra et al.,
1991; Reinhard et al., 1982). Based on the biochemical and
pharmacological properties, two main subfamilies of dopamine

0014-2999/$ - see front matter & 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.ejphar.2013.01.023
26 M. Reza Zarrindast et al. / European Journal of Pharmacology 702 (2013) 2531
receptors have been described. These include dopamine D1-like
(D1 and D5) and dopamine D2-like (D2, D3 and D4) subfamilies
(Sealfon and Olanow, 2000).
Compelling evidence have shown that cholestasis increases the
systemic opioids level (Dehpour et al., 1998; Ebrahimkhani et al.,
2006; Ghafourifar et al., 1997; Talaenko et al., 2005) and this
phenomenon is involved in the cholestasis-induced anxiolytic-like
behaviors (Eslimi et al., 2011). Given the close link between
opioidergic and dopaminergic systems with regard to anxiety-like
behavior regulation (Radke et al., 2011; Rezayof et al., 2009), it is
possible that opioidergic systems induce anxiolytic-like behaviors
through their interactions with dopaminergic systems in different
brain sites. The above interaction seems to occur within mesolimbic
projections. Activation of dopaminergic neurons of nucleus accum-
bens and ventral tegmental area, in particular, could be mediated by
the opioid peptides (Bruijnzeel, 2009; Koob and Le Moal, 2008;
Radke et al., 2011).
Referring to the previous studies which have indicated dopa-
minergic system changes in cholestatic subjects, as well as the
involvement of dopaminergic systems in regulation of anxiety-
like behaviors, the aim of the present study is to nd out the
possible involvement of dopamine D1 and D2 receptors in
cholestasis-induced anxiolytic-like behaviors in rats.
2. Materials and methods
2.1. Animals
Male Wistar rats (bred at the institute of cognitive science,
Tehran, Iran), weighing 220240 g at the time of the surgery, were
used in this study. Animals were housed in Plexiglas cages in
constant temperature (2272 1C) and automatically controlled
light/dark cycle (12:12 h light/dark). They had access to commercial
rodent pellets and water ad libitum. Four to ve animals were
housed in a same cage and eight were used in each experimental
group. Each rat was used once only. All experimental procedures
and animal use protocols were approved by the Research and Ethics
Committee of the Faculty of Science, Tehran University of Medical
Sciences.
2.2. Surgical procedure
Two experimental animal groups were used: sham-operated and
bile duct ligation. Bile duct ligation was performed under general
anesthesia induced by an intraperitoneal injection of ketamine
hydrochloride 10% (50 mg/kg) plus xylazine 2% (5 mg/kg). Briey,
the common bile duct was located through a midline abdominal
incision, double ligated near the liver, and transected between
ligatures. Sham-operated rats underwent the same procedure
except that the bile duct was manipulated without ligation or
resection (to equalize the possible stress induced by surgery in both
sham- and bile duct ligation operated groups). Animals were moved
to their cages 5 h post operation to prevent wound dehiscence.
2.3. Elevated plus maze
Elevated plus maze is an anxiety assessment test in rodents
that is used as a screening test of all currently available models of
both anxiogenic and anxiolytic agents. Elevated plus maze appa-
ratus consisted of two open arms (50  10 cm) and two closed
arms (50  10 cm) with 40-cm high walls, extending from a
central platform (10  10 cm) to make the shape of a plus sign.
The whole apparatus was elevated 50 cm above the oor. The test
room was illuminated by two 60-W bulbs located 1.5 m above the
apparatus. Animals were placed in the junction of the four arms,
after which their entries/duration in each arm as well as their
other behavioral patterns (i.e., rearings, head dip) were observed
for 5 min. The %OAT and %OAE, reect the standard anxiety
indices. Total arm entries were measured as an index for loco-
motor activity. A decrease or increase in duration of stay and
entries to open arms reect anxiogenic- and anxiolytic-like
behaviors, respectively.
2.4. Other behavioral analysis
Number of rearings (the rat maintains an erect posture which
is usually associated with snifng) and groomings (the rat rubs its
face, ears, mouth, vibrissae and eyes with rapid circular move-
ments of its forepaws), as well as the defecation index (the
number of boil defection) were considered as the conventional
indices for anxiety-like behaviors (Casarrubea et al., 2009; Eslimi
et al., 2011; Kalueff et al., 2004). Rearing, grooming and defeca-
tion data were thoroughly analyzed, although not shown here.
Experiments were performed by someone blinded to the
responses and statistical measurements.
2.5. Drugs
The drugs used in this study were ketamine and xylazine (Alfasan
Chemical Co, Woerden, Holland), apomorphine (dopamine D1 and D2
receptor agonist), SCH23390 (dopamine D1 receptor antagonist, R(1)-
7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-
enzazepine hydrochloride, Tocris, UK) and sulpiride (dopamine D2
receptor antagonist, Sigma Chemical Co., St Louis, CA, USA). All drugs
were dissolved in sterile 0.9% saline just before the experiment,
except for sulpiride which was dissolved in one drop glacial acetic
acid and sterile 0.9% saline. Control animals received saline or vehicle
(for sulpiride). All drugs were administered intraperitoneally.
2.6. Experimental protocol
Ten and 13 days post bile duct ligation, the animals behaviors
were tested in the elevated plus maze. Two groups of animals (i.e.
sham-operated and bile duct ligation) groups were considered in
each experiment.
2.6.1. Experiment 1: effects of SCH23390 on cholestasis-induced
anxiolytic-like behaviors
Eight groups of animals were used in this experiment. The
animals (sham-operated or cholestatic rats) received either intra-
peritoneal injection of saline (1 ml/kg) or different doses of
SCH23390 (0.005, 0.01 and 0.02 mg/kg), 30 min prior to testing
(13 days post bile duct ligation).
2.6.2. Experiment 2: effects of sulpiride on cholestasis-induced
anxiolytic-like behaviors
Ten groups of animals were used in this experiment. Sham-
operated or cholestatic rats received intraperitoneal injection of
saline (1 ml/kg), vehicle (1 ml/kg) or different doses of sulpiride
(0.125, 0.25 and 0.5 mg/kg), 30 min prior to testing (13 days post
bile duct ligation).
2.6.3. Experiment 3: effects of apomorphine on cholestasis-induced
changes in exploratory behaviors (10 days post bile duct ligation)
Six groups of animals were used in this experiment. Sham-
operated and cholestatic rats either received intraperitoneal injec-
tions of saline (1 ml/kg) or apomorphine at doses of 0.25 and
0.5 mg/kg, 30 min prior to testing 10 days post bile duct ligation.
 M. Reza Zarrindast et al. / European Journal of Pharmacology 702 (2013) 2531 27

2.6.4. Experiment 4: effects of SCH23390 or sulpiride on anxiolytic-

like behaviors induced by apomorphine in cholestatic rats
Nine groups of animals were used in this experiment. The
animals received intraperitoneal injection of saline (1 ml/kg), vehicle
(1 ml/kg), subthreshold doses of SCH23390 (0.005, 0.01 and
0.02 mg/kg) or sulpiride (0.125, 0.25 and 0.5 mg/kg), 30 min prior
to injection of saline (1 ml/kg) or subthreshold dose of apomorphine
(0.25 mg/kg) in cholestatic rats (10 days post bile duct ligation).
All behaviors were assessed 30 min following the last injection.

2.7. Statistical analysis

Data were represented as mean7S.E.M and the statistical

analysis of the data was made by the analysis of variance, one and
two-way ANOVA, followed by post hoc Tukeys test. Each value
displays the mean for the eight animals per group in all performed
experiments. Differences with Po0.05 between experimental
groups at each point were considered statistically signicant.

3. Results

3.1. Cholestasis induction

One day after bile duct ligation, the animals showed signs of
cholestasis (jaundice, dark urine and steatorrhea). These signs
have already been tested both qualitatively and quantitatively by
other investigators (Eslimi et al., 2011; Zarrindast et al., 2012b).

3.2. Health status

Our data indicated no alteration in body weight in

cholestatic rats (13 days post bile duct ligation, Mean7S.E.M
226.1275.7 gr) compared to the sham-operated (Mean7S.E.M
234.3875.8 gr) group.

3.3. Effects of SCH23390 on cholestasis-induced anxiolytic-like

behaviors

Two-way ANOVA and post hoc Tukeys analysis showed that 0.01
and 0.02 mg/kg of SCH23390, decreases the %OAT [dose effect:
F (3, 56)5.08, Po0.01, bile duct ligation effect: F (1, 56)0.23,
P40.05, dose-bile duct ligation interaction: F (3, 56)5.24,
Po0.01; Fig. 1A, left and right panels], while increases the number
of groomings [dose effect: F (3, 56)1.35, P40.05, bile duct ligation
effect: F (1, 56)21.68, Po0.001, dose-bile duct ligation interaction:
F (3, 56)3.93, Po0.05] and rearings [dose effect: F (3, 56)2.61,
P40.05, bile duct ligation effect: F (1, 56) 2.27, P40.05, dose-bile
duct ligation interaction: F (3, 56)4.64, Po0.01]. This however, did
not alter other behavioral indices including the %OAE [dose effect:
F (3, 56)1.06, P40.05, bile duct ligation effect: F (1, 56)0.24,
P40.05, dose-bile duct ligation interaction: F (3, 56)2.65,
P40.05; Fig. 1B, left and right panels], number of defecations [dose
effect: F (3, 56)3.27, Po0.05, bile duct ligation effect: F (1, 56)
0.87, P40.05, dose-bile duct ligation interaction: F (3, 56)1.87,
P40.05] and locomotor activity [dose effect: F (3, 56)1.12,
P40.05, bile duct ligation effect: F (1, 56)4.92, Po0.05, dose-
bile duct ligation interaction: F (3, 56)1.51, P40.05; Fig. 1C, left
and right panels] induced by cholestasis in rats (13 days post bile
duct ligation).
3.4. Effects of sulpiride on cholestasis-induced anxiolytic-like
behaviors
Two-way ANOVA and post hoc Tukeys analysis showed that 0.25
and 0.5 mg/kg of sulpiride, decreases the %OAT [dose effect: F (4, 70)
Fig. 1. Effects of SCH23390 on exploratory behaviors in sham-operated and bile
duct ligated rats. The animals received either pre-test injection of saline (1 ml/kg)
or SCH23390 (0.005, 0.01 and 0.02 mg/kg) 30 min prior to testing. The exploratory
behavior indices include: %OAT (A), %OAE (B) and the locomotor activity (C). Each
bar represents mean 7 S.E.M. P o0.01 when compared to saline in the left panel
and nnP o 0.01 when compared to saline in the right panel.
2.2, P40.05, bile duct ligation effect: F (1, 70)9.6, Po0.01, dose-
bile duct ligation interaction: F (4, 70) 6.44, Po0.01; Fig. 2A, left and
right panels] while increases the number of rearings [dose effect:
F (4, 70)1.34, P40.05, bile duct ligation effect: F (1, 70)4.20,
Po0.05, dose-bile duct ligation interaction: F (4, 70)3.29, Po0.05].
This after all, did not alter other behavioral indices including the
%OAE [dose effect: F (4, 70)3.46, Po0.05, bile duct ligation effect: F
(1, 70)6.05, Po0.05, dose-bile duct ligation interaction: F (4, 70)
1.15, P40.05; Fig. 2B, left and right panels], locomotor activity [dose
effect: F (4, 70)0.37, P40.05, bile duct ligation effect: F (1, 70)
4.31, Po0.05, dose-bile duct ligation interaction: F (4, 70)0.95,
P40.05; Fig. 2C, left and right panels], groomings [dose effect:
F (4, 70)0.05, P40.05, bile duct ligation effect: F (1, 70)17.13,
Po0.001, dose-bile duct ligation interaction: F (4, 70)0.72, P40.05]
and defecations [dose effect: F (4, 70)1.09, P40.05, bile duct
ligation effect: F (1, 70)0.07, P40.05, dose-bile duct ligation
interaction: F (4, 70)0.91, P40.05] in cholestatic rats, 13 days post
bile duct ligation.
28 M. Reza Zarrindast et al. / European Journal of Pharmacology 702 (2013) 2531

P40.05, bile duct ligation effect: F (1, 42) 2.23, P40.05, dose-
bile duct ligation interaction: F (2, 42) 0.23, P40.05], groomings
[dose effect: F (2, 42) 0.144, P40.05, bile duct ligation effect:
F (1, 42) 0.142, P40.05, dose-bile duct ligation interaction:
F (2, 42)2.31, P 40.05] and defecations [dose effect: F (2, 42)
0.30, P 40.05, bile duct ligation effect: F (1, 42) 1.03, P40.05,
dose-bile duct ligation interaction: F (2, 42) 1.29, P 40.05] in
cholestatic rats, 10 days post bile duct ligation.

3.6. Effects of SCH23390 or sulpiride on apomorphine-induced

anxiolytic-like behaviors in cholestatic rats

One-way ANOVA and post-hoc Tukeys analysis indicated that

the subthreshold dose of SCH23390 (0.01 and 0.02 mg/kg),
signicantly reduces the %OAT [F(4, 35) 8.15, Po0.001;
Fig. 4A, left panel], locomotor activity [F(4, 35) 4.08, Po0.05;
Fig. 4C, left panel] whereas, increases the number of groomings
[F(4, 35) 3.58, Po0.05] and rearings [F (4, 35)6.97, Po0.001].
This however, failed to alter %OAE [F (4, 35) 2.4, P40.05; Fig. 4B,

Fig. 2. Effects of sulpiride on exploratory behaviors in sham-operated and bile

duct ligated rats. The animals received pre-test injection of saline (1 ml/kg),
vehicle (1 ml/kg) or sulpiride (0.125, 0.25 and 0.5 mg/kg) 30 min prior to testing.
The exploratory behavior indices include: %OAT (A), %OAE (B) and the locomotor
activity (C). Each bar represents mean 7S.E.M. nnPo 0.01 when compared to saline
in the left panel and nPo 0.05 and nnPo 0.01 when compared to saline in the right
panel.

3.5. Effects of apomorphine on exploratory behaviors in sham-

operated and cholestatic rats (10 days post bile duct ligation)

Two-way ANOVA and post-hoc Tukeys analysis showed that,

apomorphine increases the %OAT [dose effect: F (2, 42) 17.72,
Po0.001, bile duct ligation effect: F (1, 42) 8.888, P o0.01, dose-
bile duct ligation interaction: F (2, 42) 1.544, P40.05; Fig. 3A,
left and right panels] and %OAE [dose effect: F (2, 42) 11.26,
Po0.001, bile duct ligation effect: F (1, 42) 8.561, P o0.01, dose-
bile duct ligation interaction: F (2, 42) 3.4, Po0.05; Fig. 3B, left
and right panels], while decreases the locomotor activity [dose
effect: F (2, 42) 10.65, Po0.001, bile duct ligation effect: Fig. 3. Effects of apomorphine on exploratory behaviors in sham-operated and
F (1, 42) 0.98, P40.05, dose-bile duct ligation interaction: bile duct ligated rats. The animals received either pre-test injection of saline (1 ml/
kg) or apomorphine (0.25 and 0.5 mg/kg) 30 min prior to testing. The exploratory-
F (2, 42) 2.73, P40.05; Fig. 3C, left and right panels]. This like behaviors include %OAT (A), %OAE (B) and the locomotor activity (C). Each bar
intervention however, did not alter other behavioral indices represents mean 7 S.E.M. nP o0.05 when compared to saline in the left panel
including number of rearings [dose effect: F (2, 42)1.67, while P o0.01 and P o 0.001 when compared to saline in the right panel.
 M. Reza Zarrindast et al. / European Journal of Pharmacology 702 (2013) 2531
left panel] and the number of defecations [F (4, 35)2.6, P40.05]
induced by the subthreshold dose of apomorphine (0.25 mg/kg) in
cholestatic rats, 10 days post bile duct ligation. In conclusion, the
data elicited that, SCH23390 blocks anxiolytic-like behaviors
induced by apomorphine in the cholestatic rats (10 days post
bile duct ligation).
Furthermore, one-way ANOVA and post hoc Tukeys analysis
showed that the subthreshold dose of sulpiride (0.5 mg/kg)
signicantly reduces the %OAT [F(5, 42)4.7, Po0.05; Fig. 4A,
right panel], the %OAE [F(5, 42) 3.3, P o0.05; Fig. 4B, right panel]
and number of rearings [F (5, 42) 9.87, Po0.001] while
increases number of groomings [F (5, 42) 5.54, Po0.001]. The
Fig. 4. Effects of SCH23390 or sulpiride on anxiolytic-like behaviors induced by
apomorphine in the cholestatic rats. The animals received pre-test injection of
saline (1 ml/kg), vehicle (1 ml/kg), SCH (0.005, 0.01 and 0.2 mg/kg) or sulpiride
(0.125, 0.25 and 0.5 mg/kg) 15 min before saline (1 ml/kg) or apomorphine
(0.25 mg/kg) injection in cholestatic rats (10 days post bile duct ligation). The
anxiolytic-like behavioral indices include changes in %OAT (A), %OAE (B) and the
locomotor activity (C). Each bar represents mean 7S.E.M. nPo 0.05 when com-
pared to saline/saline group, P o0.05, P o0.01 and P o 0.001 when com- Our results are completely in line with the previous investigations
pared to saline/apomorphine group.
29
above did not change other behavioral indices such as the
locomotor activity [F (5, 42) 1.24, P 40.05; Fig. 4C, right panel]
and number of defecations [F (5, 42) 2.02, P40.05] induced by
the subthreshold dose of apomorphine (0.25 mg/kg) in cholestatic
rats, 10 days post bile duct ligation. Our data suggested that
sulpiride blocks the anxiolytic-like behaviors induced by apomor-
phine in cholestatic rats, 10 days post bile duct ligation.
4. Discussion
4.1. Effects of cholestasis on anxiolytic-like behaviors
In agreement with our previously published data, the present
results demonstrated that cholestasis in rats (13 but not 10 days post
bile duct ligation) induces anxiolytic-like behaviors while does not
alter locomotor activity in the elevated plus maze test (Eslimi et al.,
2011). Some studies have indicated that, acute and chronic liver
failure, alter the level of activity in opioidergic (Roberts et al., 1987;
Zhang et al., 2004), dopaminergic (Glaser et al., 2006, 2003; Zimatkin
et al., 2008), cholinergic (Sundewall and Lefvert, 1990; Zarrindast
et al., 2012a), GABAergic (Ahboucha et al., 2008; Ferenci et al., 1983),
adrenergic (Borhani et al., 2005; LeSage et al., 2004), serotonergic
(Celik et al., 2005; Nguyen et al., 2008) and glutamatergic (Mendez
et al., 2009; Ohara et al., 2009) systems. Moreover, it has been
documented that alterations in the release of the corticotrophin-
releasing hormone (Bearn et al., 1995; Burak et al., 2002; Swain et al.,
1993) and the changes in brain manganese level (Forton et al., 2004)
are involved in altered cognitive and non-cognitive behaviors
induced by cholestasis. On the other hand, the mesocortical and
mesolimbic dopaminergic systems role in anxiety-like behaviors
(Nasehi et al., 2011; Talaenko et al., 2005; Zarrindast et al., 2010)
have received as much attention as the GABAergic, serotonergic and
noradrenergic systems (Hayes and Schulz, 1983; Nikolaus et al.,
2010; Noyes, 1985). Therefore, the question which raised here was
whether the post cholestasis changes in the dopaminergic system
activity level are involved in the anxiolytic-like behaviors induced by
bile duct ligation.
4.2. Effects of dopaminergic agents on cholestasis-induced
anxiolytic-like behaviors
The results indicated that, pre-test injection of apomorphine, in
sham-operated rats, increases and decreases the %OAT and the %OAE,
respectively. This however did not alter other behaviors suggesting an
anxiolytic-like effect for the drug. On the other hand, sole pre-test
injection of SCH23390 or sulpiride, at applied doses, did not alter the
%OAT, %OAE and other behaviors in sham-operated rats. Moreover,
pre-test administration of subthreshold doses of SCH23390 or
sulpiride, decreased the %OAT however did not affect other beha-
vioral indices induced by cholestasis (13 days post bile duct ligation).
The above data indicated that, both SCH23390 and sulpiride decrease
the cholestasis-induced anxiolytic-like behaviors. In addition, pre-test
injection of the subthreshold dose of SCH23390 or sulpiride before
administration of the subthreshold dose of apomorphine in chole-
static rats (10 days post bile duct ligation), decreased anxiolytic-like
behaviors induced by the subthreshold dose of apomorphine in 10
days post bile duct ligation rats. The higher doses of SCH23390 and
sulpiride however, decreased the locomotor activity and %OAE,
respectively. Based on the above, the present results strongly suggest
the involvement of dopaminergic system (dopamine D1 and D2
receptors) in cholestasis-induced anxiolytic-like behaviors. Since we
did not use selective dopamine D1 or D2 receptor agonists, further
experiments seem necessary to substantiate the above hypothesis.
showing that, both dopamine D1 and D2 receptors are involved in
30 M. Reza Zarrindast et al. / European Journal of Pharmacology 702 (2013) 2531
morphine induced anxiolytic-like behaviors (Rezayof et al., 2009).
It has been shown that, systemic injection of apomorphine decreases
the anxiety-like effects in the elevated plus maze via the dopamine
D2 receptor subtype (Garcia et al., 2005). However, our previous data
indicated that, the intra-ventral hippocampal injection of apomor-
phine induces anxiogenic-like effects (Zarrindast et al., 2010) which
may be due to the effect of apomorphine on the dopamine D2
presynaptic receptor and decrease in the dopamine level. It should be
considered that in our previous experiments dopamine receptors
antagonist decrease anxiogenic-like effect when were injected into
nucleus accumbens (Zarrindast et al., 2012c) and ventral hippocam-
pus (Zarrindast et al., 2010). However theses antagonists when were
injected peripherally reduced anxiolytic effect by bile duct ligation.
One may assume that the antagonists affected indirectly either on
release or turnover of the morphine which induced anxiolytic-like
activity. By the way our experiment data indicated that the antago-
nists have no effect on locomotor activity and this issue cannot be
involved.
Moreover, some investigations have emphasized on the dopami-
nergic systems pivotal role in gut functions such as intestinal motility
(Anlauf et al., 2003; Li et al., 2004). This has been suggested to be via
ve classes of dopamine receptors which are expressed throughout
the proximo-distal axis of the bowel (Li et al., 2006). The endogenous
dopamine for instance, decreases the intestinal motility via dopamine
D2 receptors. This indicates that, the dopamine D2 receptors are
important at all levels of the bowel function (Li et al., 2006). The fact
that dopaminergic system regulates the proliferation and secretion
mechanism(s) of cholangiocytes in the liver, supports its role in
regulation of liver function. To further elaborate on this, we may note
that: 1-following cholestasis, the functional and proliferative
responses of cholangiocytes are blocked by 6-hydroxidopamine (an
agent inducing dopaminergic cell death) (Glaser et al., 2006); 2-ductal
secretion in bile duct ligation rats are blocked by dopamine D2
receptor agonist (Glaser et al., 2003); 3-liver function failure
decreases the dopamine level in the frontal cortex, hippocampus
(Yurdaydin et al., 1990) and striatum (Zeneroli et al., 1991).
In contrast, some investigations have revealed that, hepatic encepha-
lopathy does not alter the dopamine level in most brain regions
(Bergqvist et al., 1995; Colombo et al., 1996; Hadesman et al., 1995;
Wikell et al., 1998). The notion whether the change in brain
dopamine level is involved in cholestasis, should be tested later.
Based on some solid evidence, the cholestasis-induced opioid
tone occurs through opioid receptor stimulation by endogenous
opioid peptides (Bergasa et al., 1994; Swain et al., 1992; Thornton
and Losowsky, 1988). Our previous study indicated that opioi-
dergic system (mu opioid receptor) involves in anxiolytic-like
behaviors induced by cholestasis (Eslimi et al., 2011). Moreover,
there are some investigations pointing out the role of dopami-
nergic systems in morphine induced behaviors, namely the
typical anxiolytic-like effects (Eslimi et al., 2011; Radke et al.,
2011; Rezayof et al., 2009), memory impairments (Zarrindast
et al., 2006; Zarrindast and Rezayof, 2004) and reward (Ma et al.,
2009; Narita et al., 2010). Therefore, it can be also inferred that
opioids possibly exert their anxiolytic-like effects through dopa-
mine release and dopaminergic system activation.
On the other hand, cholestasis may alter the activity of different
enzymatic pathways such as the monoamine oxidase (MAOB).
According to literature, the hippocampal activity of this enzyme
in astrocytes and radial glial cells (Mallajosyula et al., 2008) is
decreased and increased two and six weeks after cholestasis,
respectively (Zimatkin et al., 2008). It has also been found that in
thioacetamide-induced hepatic encephalopathy, the level of dopa-
mine decreases in the frontal cortex and hippocampus (Yurdaydin
et al., 1990). Moreover, in porto-systemic drug induced hepatic
encephalopathy, the level of dopamine decreases in the striatum
(Zeneroli et al., 1991).
The subthreshold dose of apomorphine (dopamine D1 and D2
receptor agonist) induces anxiolytic-like behaviors 10 days post bile
duct ligation (which did not alter anxiety-like behaviors by itself).
This response was block by SCH23390 and sulpiride. Furthermore,
the antagonists block the cholestasis-induced anxiolytic-like beha-
viors in rats; 13 days post bile duct ligation. Taken all these together,
we may conclude that the anxiolytic-like behaviors induced by bile
duct ligation are possibly mediated through dopamine D1 and D2
receptor mechanisms.
Acknowledgments
The authors thank the Iran National Science Foundation (INSF)
for providing the nancial support for the project.
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