Neurogastroenterol Motil (2006) 18, 949–956
Involvement of cannabinoid receptors in inflammatory
hypersensitivity to colonic distension in rats
M. SANSON, L. BUENO & J. FIORAMONTI
Neurogastroenterology and Nutrition Unit, INRA, Toulouse, France
Abstract Activation of cannabinoid CB1 and CB2
receptors is known to attenuate nociception and
hyperalgesia in somatic inflammatory conditions. The
aim of this study was to determine whether cannabi-
noids modulate colonic sensitivity in basal and in-
flammatory conditions. The effects of CB1 and CB2
receptor agonists and antagonists on the abdominal
contractile response to colorectal distension
(CRD) in basal conditions and after 2,4,6-trini-
trobenzenesulphonic acid-induced colitis were
investigated. As previously described, colitis triggered
a hypersensitivity to CRD. In basal conditions, both
CB1 (WIN 55212-2) and CB2 (JWH 015) agonists
reduced the abdominal response to CRD at a dose of
1 mg kg)1, i.p. Both compounds were active at a lower
dose (0.1 mg kg)1) abolishing the hypersensitivity
induced by colitis. Administered alone, CB1
(Rimonabant) and CB2 (SR 144528) receptor antago-
nists (10 mg kg)1) had no effect on basal sensitivity.
In contrast, the CB1, but not the CB2, receptor
antagonist enhanced colitis-induced hyperalgesia. It is
concluded that colonic inflammation enhances the
antinociceptive action of CB1 and CB2 receptor
agonists, and activates an endogenous, CB1 receptor
mediated, antinociceptive pathway.
Keywords cannabinoids, CB receptors, colorectal
distension, inflammatory hyperalgesia.
INTRODUCTION
Two cannabinoid receptor subtypes, both G protein-
linked, have been isolated and cloned to date. CB1
Address for correspondence
Jean Fioramonti, Neurogastroenterology and Nutrition Unit,
INRA, 180 chemin de Tournefeuille, BP 3, F-31931 Toulouse
Cedex 9, France.
Tel.: +33 561 28 51 49; fax: +33 561 28 53 97;
e-mail: jfioramo@toulouse.inra.fr
Received: 18 January 2006
Accepted for publication: 11 May 2006
Ó 2006 The Authors
Journal compilation Ó 2006 Blackwell Publishing Ltd
doi: 10.1111/j.1365-2982.2006.00819.
receptors are mostly expressed by central and periph-
eral neurones,1 but also by epithelial cells,2 while CB2
receptors are expressed mostly by immune cells.3
Endogenous lipid ligands for these receptors have been
isolated and identified. The most known are ananda-
mide4 and 2-arachidonoyl glycerol (2-AG)5,6 that can
bind both receptors.
Activation of CB receptors, mainly CB1 receptors,
is known to induce a large range of effects on the
gastrointestinal tract. For example, CB1 receptor
agonists delay gastric emptying in rats,7 inhibit the
occurrence of transient lower oesophageal sphincter
relaxations (TLOSR) induced by gastric air insuffla-
tion in dogs,8 attenuate pentagastrin-induced gastric
acid secretion in rats,9 inhibit the electrically evoked
acetylcholine and the subsequent contraction in
intestinal myenteric plexus-longitudinal muscle pre-
paration in guinea-pig,10 and reduce the ileal secre-
tion induced by electrical stimulation in rats.11 The
anandamide congener palmitoylethanolamide inhibits
small intestine motility12 and its levels are enhanced
in colon biopsies from patients with ulcerative
colitis.13
Among the roles of the endocannabonoid system, its
analgesic action at spinal and peripheral levels is
largely documented.14 However, these data are mainly
related to somatic pain and the role of cannabinoids in
the control of visceral nociception has been poorly
investigated. It has been shown that anandamide, via
CB1 receptors, attenuates hyperalgesia-induced urinary
bladder inflammation in the rat.15 Concerning the
gastrointestinal tract, putative actions of cannabinoids
on visceral perception have been proposed,16 but no
experimental data are available. Several observations
support the hypothesis of a control of gastrointestinal
perception by the endocannabinoid system. Ananda-
mide has been found to act at the receptor for
capsaicin, the transient receptor potential vanilloid
receptor 1 (TRPV1),17 which plays a key role in visceral
hyperalgesia. CB1 receptors have been found localized
on extrinsic primary afferent nerves in guinea-pig
949
M. Sanson et al.
submucosal plexus18 and in the rat dorsal root gan-
glia.19
Moreover, the role of cannabinoids seems to be
enhanced in inflammatory conditions. Besides the
anti-inflammatory effects of cannabinoid ligands,20
activation of either CB1 or CB2 receptors is able to
attenuate the induction and maintenance of inflam-
matory pain in rats.21 At the gastrointestinal level,
expression of CB1 receptors has been found to be
upregulated in rats during an experimental inflamma-
tion,22,23 and anandamide levels have been found to be
increased in 2,4,6-trinitrobenzenesulphonic acid
(TNBS)-induced colitis and in ulcerative colitis.24
Consequently, the aim of this study was to determine
whether activation of CB1 and CB2 receptors can
reduce the sensitivity to colorectal distension (CRD) in
the rat and the hyperalgesia induced by TNBS colitis. A
role of the endogenous cannabinoid system, in basal
and inflammatory conditions, was also investigated by
using selective antagonists for CB1 and CB2 receptors.
MATERIALS AND METHODS
Animals and surgical procedure
Male Wistar rats (Janvier SA, Le Genest St Isle,
France) were surgically prepared for electromyogra-
phy, according to a previously described technique.25
They were premedicated with acepromazine
(0.6 mg kg)1, Calmivet; Vetoquinol, Lure, France)
intraperitoneally (i.p.) and anaesthetized by i.p.
administration of ketamine (120 mg kg)1, Imalgene
1000; Rhône-Merieux, Lyon, France). Three pairs of
nichrome wire electrodes (60 cm in length and 80 lm
in diameter) were implanted bilaterally into the
abdominal external oblique muscle, 2 cm laterally
from the midline, just superior to the inguinal
ligament. The free ends of the electrodes were
exteriorized on the back of the neck and protected
by a glass tube attached to the skin. Rats were
individually housed in polypropylene cages and kept
in a temperature-controlled room (21 °C). They were
allowed free access to water and food (pellets; UAR
Epinay, France). All protocols were approved by the
Local Animal Care and Use Committee of Institut
National de la Recherche Agronomique.
Distension procedure and electromyographic
recordings
Rats were placed in polypropylene tunnels (diameter
7 cm, length 20 cm), where they could not move,
escape or turn around. They were accustomed to this
950
Neurogastroenterology and Motility
procedure for 3 days before any CRD, in order to
achieve familiarization with that environment.26 The
balloon used for CRD was 4 cm long and made from a
latex condom. It was fixed on a semi-rigid catheter
(2 mm in diameter). CRDs were performed by insertion
of the tip of the balloon 4 cm from the anus. The
catheter was fixed at the base of the tail and connected
to a computerized barostat.27 The balloon was inflated
progressively in steps of 15 mmHg, each step of
inflation lasting 5 min. Colorectal pressure and balloon
volume were continuously monitored on a potentio-
metric recorder (L6517; Linseis, Selb, Germany) with a
paper speed of 1 cm min)1. The electrical activity of
the abdominal striated muscle was recorded with an
electroencephalograph machine (Mini VIII; Alvar,
Paris, France) using a paper speed of 4 cm min)1, and
a short time constant (0.03 s) to remove low-frequency
signals (<3 Hz).
Experimental protocols
Experiments were performed in groups of eight rats.
Colitis was induced in seven groups. After an overnight
fast rats were treated with TNBS at a dose of
80 mg kg)1 in 0.3 mL 50% ethanol. TNBS was infused
for 30 min through a silicone rubber catheter intro-
duced 3 cm into the anus under acepromazine–keta-
mine anaesthesia, as described previously.28
Each rat was submitted to CRD 3 days before and
3 days after TNBS administration. Vehicle (10%
DMSO/water solution, 0.2 mL) or CB1 and CB2 recep-
tor agonists and antagonists were administered i.p.
20 min before CRD. Group 1 received vehicle, groups 2
and 3 WIN 55212-2 (CB1 receptor agonist) at 0.1 and
1 mg kg)1, respectively, groups 4 and 5 JWH 015 (CB2
receptor agonist) at 0.1 and 1 mg kg)1, respectively,
and groups 6 and 7 received at a dose of 10 mg kg)1,
Rimonabant (CB1 receptor antagonist) and SR 144528
(CB2 receptor antagonist) respectively. Group 8 was a
control group receiving saline instead of TNBS-eth-
anol, and vehicle of agonists and antagonists. Accord-
ing to the relative low selectivity of the CB1 receptor
agonist WIN 55212-2 for CB1 vs CB2 receptors,29 we
attempted to block the effects of WIN 55212-2 by the
highly selective CB1 and CB2 receptor antagonists,
Rimonabant and SR 144528 respectively. Similarly,
attempts were made to block the effects of the CB2
agonist JWH 015 with the CB1 and CB2 antagonists. To
assess the selectivity of the effects of the CB1 and CB2
receptor agonists these assays were performed in
supplementary groups of rats in the absence and
presence of colitis, the antagonist being administered
i.p. 20 min before the agonist.
Ó 2006 The Authors
Journal compilation Ó 2006 Blackwell Publishing Ltd
Volume 18, Number 10, October 2006 Role of cannabinoids in inflammation

15 mmHg, and these correlated with increases in

Chemicals
colonic pressure. Compared with vehicle, both the
WIN 55212-2 [R-(+)-[2,3-dihydro-5-methyl-3-[(morpho- CB1 receptor agonist, WIN 55212-2, and the CB2
linyl)-methyl]pyrrolo [1,2,3-de]-1,4-benzoazinyl]- receptor agonist, JWH 015, each at 1 mg kg)1 i.p.,
(1-naphtalenyl)methanone mesylate] and JWH 015 significantly reduced (P < 0.05) the number of abdom-
[(2-methyl-1-propyl1H-indol-3-yl)-1-cyclohexanol] were inal contractions evoked during CRD at pressures of 30
purchased from Tocris Cookson Inc. (Bristol, UK). and 45 mmHg (Fig. 1). No significant effect (P < 0.05)
Rimonabant [N-piperidino-5-(4-chlorophenyl)-1-(2,4-di- was observed at the dose of 0.1 mg kg)1 for both
chlorophenyl)-4-methyl-3- pyrazole-carboxamide] and compounds (data not shown). Both the CB1 receptor
SR 144528 [N-(1S)-endo-1,3,3,-trimethyl bicyclo antagonist, Rimonabant, and the CB2 receptor antag-
[2.2.1]heptan-2yl-5-(4-chloro-3-methyl-phenyl)-1(4- onist, SR 144528, at the dose of 10 mg kg)1 i.p., did not
methylbenzyl)- pyrazole-3-carboxamide] were gifts modify significantly (P < 0.05) the number of abdom-
from SANOFI Research Centre (Montpellier, France). inal contractions, whatever the CRD pressure, in
All compounds were dissolved in a 10% DMSO/water comparison with vehicle treatment (Fig. 1). None of
solution. 2,4,6-trinitrobenzenesulfonic acid was pur- the compounds tested modified the balloon volume,
chased from Sigma (St Louis, MO, USA). irrespective of the distending pressure.

Data analysis Inflammatory conditions

The number of spike bursts (corresponding to abdom- Three days after TNBS, the number of abdominal
inal contractions) was determined per 5-min period, contractions evoked by CRD at the pressure of
from the onset of balloon inflation to the beginning of 15 mmHg was significantly increased (P < 0.05) in
deflation. Colorectal volumes were determined on a comparison with the first CRD session performed
potentiometric recorder as the maximal volume ob- 3 days after intracolonic administration of saline
tained for each stage of distension. Statistical analysis (Fig. 2). This increase was abolished by either the
of the number of abdominal contractions and colorec- CB1 receptor agonist, WIN 55212-2, or the CB2 recep-
tal volumes was performed using a one-way analysis of tor agonist, JWH 015, at a dose of 0.1 mg kg)1 i.p.,
variance (ANOVA) followed by post hoc test. Data are which was ineffective in basal conditions. Moreover,
expressed as mean ± SEM. the number of abdominal contractions after WIN
55212-2 or JWH 015 was significantly lower (P < 0.05)
than in the absence of inflammation for the distending
RESULTS
pressure of 30 mmHg (Fig. 2). The CB1 receptor antag-
onist Rimonabant, at the dose of 10 mg kg)1 i.p., did
Basal conditions
not affect per se the basal sensitivity, but significantly
Under basal conditions (in the absence of inflamma- enhanced (P < 0.05) the increase in the number of
tion), increasing CRD progressively increased the abdominal contractions observed at the pressure of
number of abdominal contractions, starting from 15 mmHg after TNBS. On the contrary, the CB2

Vehicle
Number of abdominal contractions per 5 min

WIN 55212-2 JWH 015

60

Rimonabant SR 144528

40
Figure 1 Effect of CB1 (WIN 55212)
and CB2 (JWH 015) receptor agonists
(1 mg kg)1, i.p.), and CB1 (Rimonab- 20
ant) and CB2 (SR 144528) receptor
*
antagonists (10 mg kg)1, i.p.) on the *
number of abdominal contractions * *
induced by increasing pressures of 0
colorectal distensions. *P < 0.05 vs 0 15 30 45 60 0 15 30 45 60
vehicle treatment. Pressure (mmHg) Pressure (mmHg)

Ó 2006 The Authors

Journal compilation Ó 2006 Blackwell Publishing Ltd 951
M. Sanson et al. Neurogastroenterology and Motility

Saline TNBS
+ Vehicle + vehicle
TNBS + TNBS +
WIN 55212-2 JWH 015
Number of abdominal contractions per 5 min

60 TNBS + TNBS +
Rimonabant SR 144528 Figure 2 Reversal of 2,4,6-trinitroben-
zenesulphonic acid (TNBS)-induced
*† allodynia by the CB1 receptor agonist
40
(WIN 55212, 0.1 mg kg)1, i.p.) and
enhancement by the CB1 receptor
* * antagonist (Rimonabant, 10 mg kg)1,
† † i.p.). The CB2 receptor agonist (JWH
20 †
† 015, 0.1 mg kg)1, i.p.) also reversed
*† TNBS-induced allodynia, but the CB2
*†
† † receptor antagonist (SR 144528,
0 10 mg kg)1, i.p.) had no effect. *P <
0 15 30 45 60 0 15 30 45 60 0.05 vs vehicle, vehicle; P < 0.05 vs
Pressure (mmHg) Pressure (mmHg) TNBS, vehicle.

Vehicle
WIN 55212-2 JWH 015
Number of abdominal contractions per 5 min

WIN 55212-2 JWH 015

60
+ Rimonabant + SR 144528

WIN 55212-2 JWH 015

+ SR 144528 + Rimonabant Figure 3 Reversal of the effect of the
40
CB1 receptor agonist (WIN 55212) by
the CB1 (Rimonabant), but not the CB2
* (SR 144528) receptor antagonist. Simi-
* larly, the effect of the CB2 receptor
20
* agonist (JWH 015) was reversed by the
*
* * CB2, but not the CB1, receptor antag-
* * onist. Agonists and antagonists were
0 administered i.p. at the doses of 1 and
0 15 30 45 60 0 15 30 45 60 10 mg kg)1 respectively. *P < 0.05 vs
Pressure (mmHg) Pressure (mmHg) vehicle treatment.

receptor antagonist SR 144528, at a dose of 10 mg kg)1 (1 mg kg)1), was blocked by the CB2 receptor antagon-
i.p., did not induce any significant change in the ist, but not by the CB1 receptor antagonist (Fig. 3).
number of abdominal contractions after TNBS Identical antagonisms were observed in the presence of
(P > 0.05), irrespective of the distending pressure colitis, where agonists were tested at the dose of
(Fig. 2). The balloon volume was significantly reduced 0.1 mg kg)1 and antagonists at 1 mg kg)1 (Table 1).
(P < 0.05) after TNBS, in comparison with saline, for
distending pressures of 45 and 60 mmHg. The com-
DISCUSSION
pounds tested did not modify these reduced volumes.
Our results indicate that activation of either CB1 or
CB2 receptors reduces the basal sensitivity and the
Selectivity of CB1 and CB2 receptor agonists
colitis-induced hypersensitivity to CRD in rats, both
In the absence of inflammation, the decrease in the agonists being more active in the presence of colitis.
number of abdominal contractions induced by the CB1 However, the endogenous cannabinoid system is only
receptor agonist, WIN 55212-2, at the dose of involved in the inflammatory hyperalgesia and only
1 mg kg)1, was blocked by the CB1 receptor antagonist through CB1 receptors, as the CB1 receptor antagon-
SR 141718A (10 mg kg)1, i.p.), but not by the CB2 ist, but not the CB2, receptor antagonist, was able to
receptor antagonist SR 144528 (10 mg kg)1, i.p.). Con- enhance the abdominal response to CRD during
versely, the effect of the CB2 receptor agonist, JWH 015 colitis.

Ó 2006 The Authors

952 Journal compilation Ó 2006 Blackwell Publishing Ltd
Volume 18, Number 10, October 2006
Table 1 Selective reversal of the effect of WIN 55212-2 by
Rimonabant, and JWH 015 by SR 144528, under inflammatory
conditions
Vehicle Rimonabant SR 144528
WIN 55212-2 22.9 ± 3.3 47.4 ± 4.1* 28.2 ± 4.4
JWH 015 26.0 ± 3.6 22.3 ± 3.7 43.2 ± 4.1*
Data are reported as number of abdominal contractions during
a 5-min period of colorectal distension at 60 mmHg
(mean ± SEM, n ¼ 8). Agonists were administered i.p. at the
dose of 0.1 mg kg)1, and antagonists at 1 mg kg)1. *P < 0.05
in comparison with agonist/vehicle.
The decrease in sensitivity to CRD induced by the
CB1 receptor agonist is supported by several anatom-
ical data. CB1 receptors are expressed in the gastroin-
testinal tract, with the highest level in the stomach
and in the colon, at least in mice.30 In rats and guinea-
pigs, besides an expression in cholinergic myenteric
neurones, a close association between CB1 receptor
immunoreactivity and fibres labelled for a synaptic
protein has been described, suggesting a role in the
modulation of transmitter release. Interestingly, very
few calcitonin gene-related peptide (CGRP)- and sub-
stance P-positive neurones are co-localized with CB1
receptor immunoreactivity, suggesting independent
pathways for cannabinoid and CGRP and/or substance
P transmission at the digestive level.31 CB1 receptors
are expressed in primary sensory neurones of dorsal
root ganglia, with some degrees of co-localization with
CGRP and TRPV1 ion channel.19,31,32 Finally, CB1
receptors are expressed in the dorsal horn of the spinal
cord,33 at both pre- and postsynaptic levels.34
CB2 receptors are known to be mostly located on
immune cells, and there is no clear anatomical
evidence to support the decrease in sensitivity to
CRD by the CB2 receptor agonist in basal conditions.
In humans, CB2 receptors have been identified in
normal colonic epithelial cells,2 as well as in colonic
tissues of inflammatory bowel diseases35 and colorec-
tal carcinomas.2 CB2 receptors have been also found on
skin afferent nerve fibres,36 but there is no evidence for
CB2 expression in gastrointestinal afferents. No CB2
receptor has been identified in the mouse dorsal root
ganglia or spinal cord, in basal conditions.37 However,
there are some data available indicating an antinoci-
ceptive action of CB2 receptor activation. An analgesic
action of CB2 receptor agonists has been shown in
inflammatory or neuropathic pain, but only few data
provide evidence for these compounds in basal condi-
tions. A CB2 receptor-selective agonist, AM 1241,
produces antinociception to skin thermal stimuli.38
Ó 2006 The Authors
Journal compilation Ó 2006 Blackwell Publishing Ltd
Role of cannabinoids in inflammation
The same compound was also able to suppress C-fibre
activity in the absence of inflammation.39
We found an enhanced activity of both CB1 and CB2
receptor agonists after induction of colitis. A colonic
anti-inflammatory action of CB1 receptor has been
suggested. Expression of CB1 receptors has been found
to be upregulated in rats during an experimental
intestinal inflammation.22,23 CB1-deficient mice or
wild-type mice treated with the CB1 receptor antag-
onist Rimonabant display a stronger experimental
colitis than controls.23 At the somatic level, the
endocannabinoid palmitoylethanolamide and the syn-
thetic cannabinoid nabilone are able to reduce a
carrageenan-induced acute hindpaw inflammation.40
However, the anti-inflammatory action of cannabinoid
is not well established, and a pro-inflammatory role
has been suggested. The CB1 receptor antagonist
Rimonabant has been found to reduce an intestinal
inflammation induced by indomethacin or lipopoly-
saccharide (LPS).41 Similarly, intraluminal administra-
tion of anandamide or 2-AG causes an iletis in the
rat.42 Nevertheless, the anti-hyperalgesic action of CB1
or CB2 receptor agonists that we observed after TNBS
treatment cannot be attributed to a possible anti-
inflammatory action, according to the short delay
(20 min) between administration of the compounds
and the measurement of colonic sensitivity to disten-
sion.
An action of both CB1 and CB2 receptor activation
in inflammatory conditions, at least at the somatic
level, has been largely documented.14 The excitatory
potency of anandamide on TRPV1 receptor is increased
by inflammatory mediators such as bradykinin or
prostaglandin E2, but the consequence in terms of
hyperalgesia or hypoalgesia has not been established.43
This is in agreement with other data indicating a
pronociceptive or an antinociceptive effect of cannabi-
noids in inflammatory conditions. Local administra-
tion of anandamide increases c-fos expression in the rat
inflamed urinary bladder and contributes to the devel-
opment of hyperalgesia during cystitis.44 On the con-
trary, peripheral activation of either CB1 or CB2
receptors has been found to suppress spinal c-fos
expression induced by intraplantar carrageenan.45
Other studies clearly indicate an antinociceptive effect
of CB1 or CB2 activation in inflammatory hypersensi-
tivity.39,46 We have shown that the efficacy of CB1 and
CB2 agonists was greater in reducing the abdominal
response to CRD during colitis than in the basal state.
Interestingly, a similar enhanced activity has been
described for the CB2 receptor agonist AM 1241 in
reducing C-fibre activity induced by stimulation of
carrageenan-inflamed paw, in comparison with the
953
M. Sanson et al.
non-inflamed paw.39 At the level of the gastrointesti-
nal tract, the effects of CB receptor agonists have been
found enhanced in inflammatory conditions. For
example, the CB1 receptor agonist CP 55940 is more
active in delaying intestinal transit in croton oil-
treated mice, where CB1 receptors have been found
to be upregulated, than in controls.22 Similarly, the
CB2 receptor agonist JWH 133 increases gastrointesti-
nal transit in LPS-treated rats, but does not modify
basal transit.47
Our results indicate that endogenous cannabinoids
are involved through CB1 receptors as a feedback
mechanism limiting colitis-induced hypersensitivity,
as in these conditions the CB1 receptor antagonist
Rimonabant administered alone enhances the abdom-
inal response to CRD. This is in agreement with a
recent finding showing that anandamide, but not
2-AG, levels are strongly increased in the colon
submucosa of TNBS-treated rats and in biopsies of
patients with ulcerative colitis.24 This increase in
anandamide level, which displays a very low efficacy
at CB2 receptors,29 and the upregulation of CB1
receptors during intestinal inflammation,22,23 may
explain the effect of the CB1 receptor antagonist in
inflammatory conditions. On the other hand, it has
been recently shown in mice that activation of TRPV1
receptors reduces the inflammation-induced irritation
of colonic smooth muscle activities.48 It can be
speculated that the abdominal response to CRD after
TNBS may be modulated through TRPV1 receptors in
two ways. If the elevated anandamide levels associated
with TNBS-induced colitis still activate TRPV1 recep-
tors when CB1 receptors are blocked, the enhancement
of the abdominal response to CRD induced by Rimo-
nabant may be underestimated. On the contrary, high
doses of Rimonabant have been found to inhibit some
TRPV1-mediated effects of anandamide.49 A blockade
of TRPV1 receptors by Rimonabant may explain the
enhanced abdominal response.
Using the antagonist Rimonabant, an involvement
of CB1 receptors has also been shown in the regulation
of gastrointestinal functions, in the absence of inflam-
mation. For example, Rimonabant enhances the cho-
linergic and NANC contractile response of the guinea-
pig ileum elicited by electrical stimulation,50 but does
not affect the response of the human ileum.51 It
increases defecation and accelerates upper gastrointes-
tinal transit in basal conditions in rats,52 and increases
the occurrence of TLOSR induced by gastric air
insufflation in dogs.8 On the contrary, it does not
modify gastric emptying in rats.7 The endogenous
involvement of CB1 receptors in the attenuation of
hyperalgesia to CRD during TNBS-induced colitis is in
954
Neurogastroenterology and Motility
agreement with their overexpression observed in the
same inflammatory model, at least on myenteric
neurones,23 and in croton oil-induced inflammation.22
Interestingly, CB1 receptors have been found to be
expressed and functional in mast cells, which are
involved in several models of visceral hyperalgesia.53
Moreover, nerve growth factor (NGF) is known to play
a key role in the colonic inflammatory hyperalgesia,54
and to maintain visceral hyperalgesia over long peri-
ods.55 CB1 receptors are expressed in NGF-responsive
neurones, but their expression is not regulated by
NGF.56 On the contrary, visceral hyperalgesia induced
by NGF (local application of NGF in the urinary
bladder) is attenuated by activation of CB1 receptors.57
Another possible link between CB1 receptors and
inflammatory nociception can be found at the level of
afferent colonic neurones expressing TRPV1 receptors,
which are sensitized by inflammatory mediators.58
CB1 and TRPV1 receptors have been found co-localized
in rat dorsal root ganglia,19 anandamide is able to act at
TRPV1 receptors,17 TRPV1 receptors have been pro-
posed as a molecular target for the antihyperalgesic
action of CB1 agonists in a rat model of acute somatic
inflammation,59 and CB1 receptor antagonists have
been found to inhibit some TRPV1 receptor-mediated
effects of anandamide.49
Finally, our results have shown an enhanced activity
of CB1 and CB2 receptors, and an endogenous involve-
ment of CB1 receptors in the attenuation of inflamma-
tory colonic hyperalgesia, but do not permit to identify a
site of action. A body of evidence suggests an action at
the level of afferent neurones, dorsal root ganglia or
dorsal horn of the spinal cord, but an action at the level of
the central nervous system cannot be excluded.
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