Digestive Diseases and Sciences, VoL 40, No. 9 (Septenlber 1995), pp.

1913--1921

Influence of Bradykinin in Gastrointestinal

Disorders and Visceral Pain Induced by
Acute or Chronic Inflammation in Rats
VI~RONIQUE JULIA, MS, THI~ODORA MEZZASALMA, MS, and LIONEL BUI~NO, DSc

This work investigated the role of bradykinin in viscerosensitivity before and during inflam-
mation in two models of visceral pain induced by rectal distension (RD) or "abdominal
distension" (AD) in rats. RD induced both inhibition of colonic motility and an increase of
abdominal spike bursts. Bradykinin receptor antagonist, Hoe 140 did not affect any of the
RD-induced responses. After TNB-induced rectal inflammation, colonic inhibition and the
number of abdominal contractions were enhanced. Hoe 140 selectively reduced the abdom-
inal response to the highest distension volume, without affecting the colonic response. In AD
group, acetic acid inhibited gastric emptying and increased the number of abdominal
contractions, whereas the same volume of saline did not affect any of the responses. Before
inflammation, Hoe 140 (1-5 mg/kg, intraperitoneally) did not affect p e r se abdominal and
gastric emptying responses; in contrast, at 5 mg/kg, intraperitoneally, it reduced significantly
(P < 0.05) both acetic acid-induced responses. We conclude that bradykinin is involved in
viscerosensitivity changes related to abdominal and rectal distension in inflammatory
conditions.

KEY WORDS: rectal distension; motility; gastric emptying; rat; inflammation; visceral pain.

Bradykinin (BK), a nonapeptide of the kinin family,
plays a major role in various inflammatory processes
such as edema (1), vasodilatation (2), and is gener-
ated in damaged tissues during inflammatory states
(3, 4). In several animal models, it has been shown
that BK is involved in the mediation of pain and
hyperalgesia caused by irritant substances (5, 6).
Moreover, application of BK to a blister base causes
pain in human volunteers (7, 8).
The biological actions of BK are mediated via two
receptor types named B1 and B2 (4). However, BK
has a higher affinity for the B2 receptor than for the
B1 receptor, and the later is known to bind
Manuscript received January 12, 1995; revised manuscript re-
ceived May 2, 1995; accepted May 8, 1995.
From the Department of Pharmacology, INRA, Toulouse,
France.
Address for reprint requests: Dr. L. Budno, Department of
Pharmacology, INRA, 180 chemin de Tournefeuille, BP 3, 31931
Toulouse cedex, France.
[desArgg]BK, an active metabolite of BK (9). Dray
and Perkins (10) suggested that B2 receptors play a
more significant role in the mediation of pain during
the earlier stages of inflammation, whereas B1 recep-
tors are likely to be involved in chronic inflammation
by maintaining an hyperalgesic state. Using autora-
diographic techniques, Steranka et al (11) showed
that BK receptors are localized on nociceptive sen-
sory neurons corresponding to small diameter unmy-
elinated fibers with cell bodies in the dorsal root
ganglia that terminate in the superficial layers of the
dorsal spinal cord. The advent of a novel B2 receptor
antagonist, Hoe 140, (12, 13) has allowed study of the
involvement of BK and B2 receptor in the genesis of
hyperalgesia related to tissue inflammation in differ-
ent animal models. However, the role of BK and of
B2 antagonist receptors in the mediation of visceral
pain is unknown.

Digestive Diseases and Sciences, Vol. 40, No. 9 (September 1995) 1913
11163-21lb/95i09(Rl-19135117.50/09 1995Plcnun*tPublishingCorporation

Several approaches have been validated to evaluate
viscerosensitivity and pain, and different criteria such
as cardiovascular reflexes or behavioral changes have
been used to evaluate pain threshold and intensity
(14). Recently, we developed a model of rectal dis-
tension ( R D ) for assessment of viscerosensitivity
c h a n g e s in awake rats using e l e c t r o m y o g r a p h i c
records from the colonic and abdominal muscles,
which allows the examination of two responses re-
lated to viscerosensitivity: (1) an inhibition of proxi-
mal colon motility and (2) an increase in the fre-
quency of abdominal cramps as an index of visceral
pain (15). Moreover, this model allows the study of
changes in both abdominal and colonic responses
during rectal inflammation induced by local applica-
tion of trinitrobenzene sulfonic acid (TNB) in ethanol
(50%) (15). A n o t h e r model o f hyperalgesia is associ-
ated with pain and inhibition of gastric emptying by
intraperitoneal injection of acetic acid (16). This
model also may be considered as a suitable prepara-
tion to evaluate the role of visceral afferents in pain
and inhibitory m o t o r reflex during the early stage of
inflammation (peritonitis). Consequently, the aim of
this work was to determine whether B2 receptors and
bradykinin play a role in viscerosensitivity changes
and the hyperalgesia induced by either acute or
chronic inflammation in two different experimental
models.
M A T E R I A L S AND METHODS
Rectal Distension
Animal Preparation. A first group of eight male Wistar
rats (Le Gen6t Saint Isle, France), weighing 250-350 g and
housed individually, was used. Animals were premedicated
with 0.3 ml of acepromazine (0.5 mg/kg) injected intraperi-
toneally and anesthetized by intramuscular administration
of 0.3 ml of ketamine (Imalg6ne 1000, Rh6ne M6rieux,
Toulouse, France). They were prepared for electromyo-
graphic recordings using a previously described technique
(17). Three groups of nichrome-wire electrodes (60 cm in
length and 80/xm in diameter) were implanted: two on the
proximal colon (2 and 3 cm from the cecum), and one in the
striated muscle of the abdomen at 3 cm laterally from the
site of laparotomy (15). In all animals, the free end of
electrodes were exteriorized on the back of the neck and
were protected by a glass tube attached to the skin. In
addition, a silicone (Silastic) catheter (30 cm) was inserted
into the intraperitoneal cavity and exteriorized together
with the electrodes. Then, the animals were allowed to
recover for eight days before the beginning of experiments.
Distension Procedure. Rectal distension (RD) was per-
formed before and three days after TNB/ethanol adminis-
tration. Three days before distension, rats were accustomed
to staying in tunnels, in order to prevent artifacts due to
1914
JULIA ET AL
movement during distension. An arterial embolectomy
catheter (Fogarty, Edwards Laboratories, Inc., Santa Ana,
California) used for distension was introduced into the
rectum at 1 em from the anus and fixed at the base of the
tail. The balloon (2 mm diameter; 2 cm long) was increas-
ingly inflated with water starting from 0.4 ml up to 1.6 ml.
Each step of inflation (0.4 ml) lasted 5 min, which was the
minimal duration permitting a relevant measurement of an
eventual colonic motor inhibition. At the end of the disten-
sion, the measurement of the volume of water withdrawn
permitted to detect any leakage.
Rectal Inflammation. Rectal inflammation was per-
formed by injection of trinitrobenzene sulfonic acid (TNB)
at a dose of 80 mg/kg (in 0.3 ml of ethanol 50%) through a
silicone catheter inserted into the rectum at 1 cm from the
anus (15).
Hoe 140 dissolved in saline (NaCI 9%c~), was infused
intraperitoneally, from 15 rain before rectal distension until
its end (ie, 30 min), at a rate of 0.15 mg/kg/min, before or
after rectal inflammation.
Abdominal Distension
Animal Preparation. A second group of rats was pre-
pared for abdominal distension (AD). In this AD group,
only two pairs of electrodes were implanted into the striated
muscles of the abdomen as described for the RD group.
Gastric Emptying Measurements. Gastric emptying was
evaluated by gavage with 2 ml of test meal containing 1
/,tCi/ml of [- Cr]sodmm chromate. Thirty minutes after the
51 9 - .
test meal, animals were killed by cervical dislocation. The
stomach and small intestine were excised, and the small
intestine was divided into 10 segments of equal length. Each
of the pieces, as well as the stomach, were placed into tubes
and radioactivity was determined in a gamma counter (MR
252C, Kontron, Basel, Switzerland) for 5 min. Gastric emp-
tying was expressed as the percentage of the administered
marker found in the small intestine.
Peritonitis Induction. Peritonitis was induced by intra-
peritoneal injection of acetic acid (0.6%) at a dose of 10
ml/kg, 10 min before the test meal.
Hoe 140 (1 and 5 mg/kg) or its vehicle (NaCI 9%e) was
injected intraperitoneally, 15 min before the test meal, ie, 5
min before intraperitoneal administration of 10 ml/kg of
acetic acid 0.6% or saline (NaCI 9% ). In addition, Hoe 140
(5 mg/kg intraperitoneally) was administered 5 min after
acetic acid 0.6% (10 ml/kg intraperitoneally) and 5 min
before test meal.
The doses used (1 and 5 mg/kg intraperitoneally) were in
agreement with studies establishing their efficacy in perito-
nitis in mice (18) and in hyperalgesia induced by adminis-
tration of carrageenin or LPS in rat paw (19).
Motility Recordings
Electromyographic recordings started eight days after
surgery. Electrical activity of the colon and abdominal con-
tractions were recorded with an electroencephalograph ma-
chine (Reega VIII, Alvar, Paris, France) at a paper speed of
2.4 cm/min. In both the RD and AD groups, electromyo-
graphic recordings started 30 min before the injection of the
antagonist. A short time constant of amplification (0.03 sec)
was used to record selectively spike bursts.
Digestive Diseases and Science's, Vol. 40, No. 9 (September 1995)
BRADYKININ AND VISCEROSENSITIVITY

TABI.E 1, EFFECT OF RECTAl. DISTENSION ON ABDOMINAL AND distension; the number of spike bursts during Hoe
COLONIC RI-SPONSES BEFORE AND AFTER RECTAL INFLAMMATION
140 infusion was not significantly different from that
Colon Abdomen observed under vehicle administration (Figure 2A).
Distension After TNB After TNB Inflamed Animals. Three days after intrarectal ad-
volumes (ml) Before (80 mg/kg) Before (80 mg/kg) ministration of TNB (80 mg/kg in ethanol 50%), the
Control 7.3 _ 0.2 6.8 _+ 0.7 0.1 • 0.1 3.3 +- 1.4"
colonic inhibition induced by rectal distension was
0.4 7.1 _+ 0.5 6.8 + 0.7 4.7 _+ 1.47 7.8 +_ 1.6"? enhanced. At a distension volume of 0.8 ml, the
0.8 6.4 • 0,4? 4.8 _+ I).5"1 1(I.4 _+ 1.37 14.9 + 1.2"? number of colonic spike bursts per 5 rain was signif-
1.2 5.4 __+0.5? 4.3 +_ (/.77 14.9 +_ 2.3? 15.6 _+ 2.2?
icantly lower (P < 0.05, N = 8) than that obtained at
1.6 4.7 = 0.4I 4.0 +_ 0.5? 17,0 + 2.5? 18.6 • 2.7?
the same volume of distension before inflammation
*P < 0.05 significantly different from c o r r e s p o n d i n g value before (Table 1). Hoe 140 (5 mg/kg intraperitoneal infusion)
T N B (N = 8).
? P < 0,05 significantly different from control values (N = 8). did not significantly change the effect of rectal disten-
sion after TNB-induced inflammation at any volume
All animal procedures included in the present study were of distension (Figure 1B).
approved by the local INRA Animal care and use Commit- Rectal inflammation induced a significant increase
tee. in the number of abdominal spike bursts compared
Chemicals with controls for the two lowest volumes of distension
(ie, 0.4 and 0.8 ml; Table 1: P < 0.05, N = 8). Hoe 140
Hoe 140 (D-Arg-[Hyp ~, Thi 5, D-TicTOicX]BKwas kindly
(5 mg/kg intraperitoneal infusion) significantly re-
provided by Dr K. J. Wirth (Hoesch, Frankfurt, Germany).
Acetic acid (98%) was purchased from Prolabo (Paris, duced the number of abdominal spike bursts for all
France). Hoe 140 and acetic acid were dissolved in saline. the volumes of distension higher than 0.4 ml (ie, 0.8
and 1.6 ml; Figure 2B).
Statistical Analysis
Statistical analysis of the number of colonic and abdom-
inal spike bursts occurring during each 5-rain period during Abdominal Distension
rectal distension was performed using analysis of variance Control Rats. Intraperitoneal injection of saline
(ANOVA) followed by Student's t test, adapted for paired
values. In the AD group, comparison of abdominal spike (10 ml/kg) did not affect gastric emptying, which was
bursts, occurring from the time of acetic acid injection until not significantly different from untreated animals
animal sacrifice, in periods of 5-min, was analyzed using t (51.9 _+ 2.8% vs 52.3 ___ 3.8%; Table 2). Hoe 140 (1
test for unpaired values. Similarly, gastric emptying values and 5 mg/kg intraperitoneally) did not influence sig-
were compared using analysis of variance (ANOVA) fol- nificantly the percentage of gastric emptying observed
lowed by unpaired Student's t test. Values were expressed
as mean -+ SEM and differences were considered significant during AD with saline (Figure 3A).
for P - 0.05. Similarly, the number of abdominal spike bursts
after intraperitoneal distension with saline (10 ml/kg)
RESULTS was not significantly different from that observed in
control animals (Table 2). Hoe 140 (1 and 5 mg/kg
Rectal Distension intraperitoneally) did not affect significantly the num-
Control Animals. In the colon, rectal distension ber of abdominal contractions (Figure 4A).
inhibited the frequency of colonic spike bursts. The Inflamed Rats. Compared with an isovolumic ad-
number of colonic spike bursts, which was 7.3 _ 0.2 ministration of saline, intraperitoneal acetic acid ad-
per 5 rain before distension, decreased to 4.7 _+ 0.4 ministration (10 ml/kg) significantly reduced (P <
for the greatest volume of distension ie, 1.6 ml (Table 0.05, N = 8) the percentage of gastric emptying (27.7
1). Hoe 140 (5 mg/kg intraperitoneal infusion) did not _+ 3.6% vs 51.9 _+ 2.8%; Table 2). Administered
significantly affect the inhibition of colonic motility before acetic acid, Hoe 140 (5 mg/kg intraperitone-
induced by rectal distension regardless of the volume ally) attenuated the acetic acid-induced slowing of
of distension used (Figure 1A). gastric emptying, which was 45.8 _ 4.4% (P < 0.05, N
In the abdomen, rectal distension increased the = 8) (Figure 3B). At a lower dose (1 mg/kg intra-
frequency of abdominal spike bursts. At the maximal peritoneally), Hoe 140 did not change significantly
volume of distension, ie, 1.6 ml, the number of ab- the inhibitory effect of IP injection of acetic acid on
dominal spike bursts reached 17.0 _+ 2.6 per 5 min gastric emptying (23.7 _ 3.1%). In contrast, when
(Table 1). Hoe 140 (5 mg/kg intraperitoneal infusion) administered after intraperitoneal injection of acetic
did not change the abdominal response to rectal acid 0.6%, Hoe 140 (5 mg/kg intraperitoneally) did

Digestive Diseases and Sciences. Vol. 40, No. 9 (September 1995) 1915
 JULIA ET AL

Before inflammation After rectal inflammation

8 8 z
r- A I I al e-
o~ 3
E o"
r
m 7 7 "o
.....IN
L_
r

J~
o
o
m 6 6 =m .
r

m m
(J
E
'-
o
5 s g
o
o r"
r

"6
4 r
B
m 3
E
z 3 I I I I I I I I I I I I

0,0 0,4 0,8 1,2 1,6 2,0 0,0 0,4 0,8 1,2 1,6 2,0
Volume (ml) Volume (ml)

-- Saline

i
w
HOE 140 5 mg/kg IP infusion
Fig 1. Effect of intraperitoneal infusion of Hoe 140 on the frequency of colonic spike bursts before (A) and after rectal
inflammation (B) (mean -2_SEM:N = 8). Values correspond to the number of spike bursts occurring during a 5-min period at each
volume of distension. *Significantlydifferent from control value (P < 0.05).

not prevent the inhibition of gastric emptying (18.3 __+
4.3% vs 27.7 ___ 3.6; Figure 5A).
Abdominal Response. Administration of acetic acid
(10 ml/kg intraperitoneally) increased significantly (P
< 0.05, N = 8) the number of abdominal spike bursts
from 5 to 35 min when compared with a similar
volume of saline (Table 2). When injected before
acetic acid, Hoe 140 (5 mg/kg intraperitoneally) sig-
nificantly reduced (P < 0.05, N = 8) the increase of
abdominal spike bursts induced by IP administration
of acetic acid, but only for the two first periods of 5
min (Figure 4B). In contrast, at the lower dose (1
mg/kg intraperitoneally), Hoe 140 was unable to in-
fluence the abdominal response to intraperitoneal
acetic acid (Figure 4B). As for gastric emptying, Hoe
140 (5 mg/kg intraperitoneally) injected 5 min after
acetic acid administration did not significantly reduce
1916
the number of abdominal spike bursts whatever the
5-rain period considered from 5 to 35 min after acetic
acid (Figure 5B).
DISCUSSION
This work shows that blockade of bradykinin B2
receptor type affects both the gastric inhibitory reflex
and visceral perception only if it is performed after
induction of rectal inflammation, suggesting that BK
does not play an important role in viscerosensitivity
under physiological conditions. Furthermore, these
effects seem to depend upon the experimental model
used.
In the first series of experiments, the effects of B2
receptor antagonist Hoe 140 on the threshold and
degree of visceral pain and colonic motor inhibition
Digestive Diseases and Sciences, Vol. 40, No. 9 (September 1995)
BRADYKININ AND VISCEROSENSITIVITY

Before inflammation After rectal inflammation

A B
20 20
Z

._c 3
o"

(/)
/ 15
D"
0
.o
3

'~ 10 10

!-

5
"6

E
Z
0 I I I I I I I I I I I 0
0,0 0,4 0,8 1,2 1,6 2,0 0,0 0,4 0,8 1,2 1,6 2,0
Volume (ml) Volume (ml)

--
i
Saline

v" HOE 140 5 mg/kg IP infusion

* p<0,05 significantly different from the control value n=8

Fig 2.Influence of intraperitoneal infusion of Hoe 140 on the frequency of abdominal spike bursts before (A) and after rectal
inflammation (B) (mean _+ SEM; N = 8). Values correspond to the number of spike bursts occurring during a 5-rain period at each
volume of distension. *Significantly different from control value (P < 0.05).

induced by rectal distension before and after inflam-

mation (15) was tested. The B2 receptor antagonist TABLE 2. EFFECT OF ABDOMINAL DISTENSION ON ABDOMINAL
AND GASTRIC EMPTYING RESPONSES (SALINE VS ACETIC ACID)
did not affect either colonic or abdominal response to
rectal distension in control animals, suggesting that Saline (10 Acetic acid
Time (rain) Control ml/kg 1P) (10 ml/kg IP)
bradykinin, or at least B2 receptors are not involved
in the normal sensitive pathways. BK is known to Number of abdominal contractions
5 9.5 +_ t.7 9.7 _+ 2.t 16,3 _+ I.I*
stimulate the release of sensory neuropeptides in
10 10.8 _+ 2.4 10.7 +_ 1.4 14.5 _+ 1.2"
various tissues (20); the vascular extravasation caused 15 9.0 _+ 2.3 6.5 -+ 1.5 12.5 +_ 1.5"
by local application of BK on nasal mucosa is dose- 20 7.3 + 2.1 4.8 +_ 1.7 13.2 + 2.0*
25 4.2 -+ 0.8 3.8 +_ 1.7 10.0 _+ 1.5"
dependently prevented by tachykinin NK1 receptor
30 4.4 + 0.9 3.3 + 1.3 10.0 -+ 0.9*
antagonist, indicating that this vascular response to 35 2.0 + 1.7 2.3 + 1.8 8.6 -+ 1.7"
BK is mediated indirectly through the release of Gastric emptying
30 51.9 + 2.7 52.3 -+ 3.8 27.7 -+ 3.6*
substance P, which increases vascular permeability
through NK1 receptors (21). Tachykinins are differ- *P < 0.05 significantly different from the control value (N = 8).

Digestive Diseases and Sciences, VoL 40. No. 9 (September 1995) 1917

Saline 10 ml/kg Acetic acid 10 mlkg
70"
I A ~i
60-
== T ~-- T to the brain. BK may be also a selective releaser of
E= ;,;,; ,;,;,
ca 4 0 -
+ seems more selectively involved in pain perception
30-
i!ii! ::::: !!ili
20"
10-
Saline Hoe 140 Hoe 140 Saline Hoe 140
lmg/kg 5mg/kg 1 mg/kg
IP IP IP
[] Saline (10ml / kg IP)
[] Acatlc acid (10 ml / kg IP)
+ significantly different from corresponding control values (p<0.05. n=8)
9 significantly d~srent from acetic acid treated rats + saline (p<0.05. n=8)
Fig 3. Effect of intraperitoneal administration of Hoe 140 on the
gastric emptying during abdominal distension before (A) and aftcr
peritoneal inflammation (B) (mean _+ SEM: N = 8). *Signiticantly
differcnt from control value (P < 0.05).
entially involved in the mediation of both colonic and
abdominal responses to rectal distension in rats. In-
deed, NK1 receptors mediate the rectocolonic inhib-
itory reflex, whereas NK2 receptors are involved in
the mediation of visceral pain (22). It may be con-
cluded that, in rectal distension, tachykinins act inde-
pendently of the BK influence since BK and B2
receptor are not involved in either response induced
by rectal distension.
As previously described, rectal inflammation in-
creases the sensitivity to rectal distension for both
inhibitory reflex and pain (15). After rectal inflamma-
tion, the B2 receptor antagonist considerably reduces
visceral pain but did not affect the rectocolonic inhib-
itory reflex. Hoe 140 reduces the effects of inflamma-
tion in different models due to its B2 receptor antag-
onist properties (6, 23). Moreover, BK and B2
receptors are present in the colon (24) and seem
involved in its physiopathology. Indeed, BK via B2
receptors activates CI- secretion in the rat colon (25)
similarly to neurokinin A in guinea pig colon (26).
Several hypotheses can be proposed to explain such a
dichotomy between the nature of the mediators in-
volved in rectocolonic inhibitory reflex and pain per-
ception, already described for the effects of NK1 and
NK2 receptor agonists (22). One may speculate that
BK selectively sensitizes silent nociceptive fibers or
1918
JULIA ET AL
that BK released from the inflammatory site may
B I
diffuse into the blood stream to reach spinal or su-
praspinal structures to amplify nociceptive messages
neurokinin A in inflamed tissue, rather than sub-
stance P, and neurokinin A through NK2 receptors
resulting from rectal distension (22).
Abdominal distension under either saline or Hoe
140 did not affect either gastric emptying or the
number of abdominal contractions, suggesting that it
does not correspond to a noxious stimulus. In con-
Hoe 140 trast, the B2 receptor antagonist partially reverses the
5 mg/kg
IP gastric emptying inhibition and abdominal responses
to intraperitoneal administration of acetic acid, sug-
gesting that bradykinin and the B2 receptor antago-
nist are involved in the genesis of noxious stimuli
resulting from acetic acid-induced peritoneal inflam-
mation, as has been observed in the RD model.
However, these effects are of short duration, since the
B2 receptor antagonist reduced the number of ab-
dominal spike bursts only during the first 10 min. This
short duration cannot be explained by Hoe 140 deg-
radation since this new B2 receptor antagonist is
known to be highly stable against enzymatic degrada-
tion and it is not a substrate for kininase II and
carboxypeptidases (13). Moreover, the fact that, at
the same dose (5 mg/kg), the B2 receptor antagonist
is active when administered before but not after ace-
tic acid suggests that BK is the first mediator involved
in peritonitis, rapidly triggering the release of other
active substances. The fact that nonsteroidal antiin-
flammatory drugs are active against acetic acid-
induced slowing of gastric emptying suggests that
inflammatory mediators are involved in this effect
(16). In contrast to the RD model, Hoe 140 is active
in reversing the inhibition of the visceromotor com-
ponent resulting from acetic acid-induced peritonitis.
In TNB-treated animals, RD is performed three days
after TNB injection, and we therefore speculate that
bradykinin plays a more selective role in pain percep-
tion in the chronic inflammatory phase (rectal inflam-
mation), while it has a major specific role in the
sensitization of primary afferents in the acute phase of
inflammation. Several studies have shown that BK
receptor antagonists have antinociceptive effects
against BK-induced and urate-induced hyperalgesia
(11) and, similar to our results, in acetic acid-induced
abdominal constrictions in mice (5). More recently,
Heapy et al 1993 (18) showed that Hoe 140 produces
a 70% inhibition of acetic acid-induced abdominal
constrictions. They suggest that BK is a major and
Digestive Diseases and Sciences, |/ol. 40, No. 9 (September 1995)
BRADYKININ AND VISCEROSENSITIVITY

Saline (10 ml / kg IP) Acetic acid (10 ml / kg IP)

2O 2O
I A I I BI z
e.
E 18 18 3
g~ T O"

16 I -
16 0.9,.

,-, 14 O"
14 ,',
0
3
"m 12 12 5"
t~
.E 10 10 -~X"
E
0 0
9oe~ 8 8 mr
e-
Ill
"6 6 6 tn
r
E 4 4 3
B ,

z
2 2

0 I 1 I I I I I I 0

0 10 20 30 40 0 10 20 30 40
Time (min) Time (min)

m
9. Saline

A
W Hoe 140 (5 m g / k g IP)

A Hoe 140 (1 mg/kg IP)

* p<0.05 significantly different from the corresponding control values (n=8).

Fig 4. Influence of intraperitoneal administration of Hoe 140 on the frequency of abdominal spike bursts during abdominal distension
before (A) and after peritoneal inflammation (B) (mean _+ SEM; N = 8). Values correspond to the number of spike bursts occurring
during a 5-rain period. *Significantly different from control value (P < 0.05).

potent, but not the only, mediator of abdominal re-
sponse and is restricted to the early phase of perito-
nitis, which is in agreement with our present results.
Moreover, Davis and Perkins (27) showed a greater
involvement of B1 receptors than B2 receptors in
development of mechanical hyperalgesia induced by
cytokines suggesting a greater role of [desArgg]BK
than BK in this model.
Our results strongly suggest that, in the two disten-
sion models used, BK is involved in pain transmission
only during inflammatory states. The involvement of
BK as an effective agent has been largely demon-
strated in different inflammatory models. BK induces
a release of different inflammatory agents such as
TNF-o~ and IL-1/3 in cultures of murine macrophage
cell lines (29). Moreover, excitation and sensitization
of articular mechanonociceptors induced by BK are
potentiated by coadministration of prostaglandin
(PG) E2 (30), and prostanoid EP 3 receptors are in-
volved in the PGE2-induced sensitization of the BK
responses of visceral nociceptors (31). Moreover
prostanoid EP 2 receptors are also involved in PGE 2

Digestive Diseases and Sciences, Vol. 40, No. 9 (September 1995) 1919
 JULIA ET AL

25 70
i- A

r 60
20
r- 50
J~
D.
15 E
r 40
U
c
'r: r
o O~ 30
"~ 10

20
,,,,
E
-t
S
Z 10

| | | | 0
i:i:i:?
10 20 30 4O
T i m e (min)
[] Acetic A c i d 0.6% (10ml/kg IP)
Acetic A c i d 0,6% (10ml/kg IP)

Hoe 140 (Smg/kg IP) injected Hoe 140 (Smg/kg IP) injected
before IP administration of Acetic [] before IP administration of Acetic
Acid (10ml/kg IP) Acid (10ml/kg IP)

,r H o e 140 (5mg/kg IP) Injected [] Hoe 140 (Smg/kg IP) injected

a f t e r IP a d m i n i s t r a t i o n of after IP administration of
Acetic A c i d (10ml/kg IP) Acetic Acid (10ml/kg IP)

~ p<0.05 significantly different from the acetic acid values (n=8).

Fig 5. Comparison of intraperitoneal administration of Hoe 140 (5 mg/kg intraperitoneally) before and after acetic acid on the fre-
quency of abdominal spike bursts (A) and gastric emptying (B) (mean • SEM; N = 8). Values correspond to the number of spike
bursts occurring during a 5-min period (A) and percentage gastric emptying (B). *Significantly different from control value (P < 0.05).

induced-sensitization of heat responses of polymodal
receptors (32) and the production of PGE 2 induced
by BK in gingival fibroblasts is coupled to an increase
in [Ca2+]i and is mediated by the B2 receptor (33). BK
is synthetized in damaged tissues during inflammatory
processes (4). Consequently, we can hypothetize that
in both RD and AD models, blockade of B2 receptor
inhibits PGE2 synthesis, which is responsible for the
sensitization of visceral nociceptors. The participation
of BK and kinin receptors in inflammatory process is
largely described. For example, the inflammatory
edema associated with the late phase of formalin-
induced pain is mediated by endogenous BK via ac-
tivation of B2 receptors (6), and Hoe 140 has been
proposed to prevent edema in acute pancreatitis (34).
However, the short time latency of Hoe 140 efficacy in
our model is probably unrelated to such antiinflam-
matory properties.
In summary, whatever the model used, BK appears
to be involved in the mediation of visceral pain during
both early and late stages of inflammation, tn con-
trast, BK or at least B2 receptors are not involved in
colonic disorders induced by rectal inflammation, but
contribute to the gastric emptying inhibition induced
by peritoneal inflammation, suggesting that the BK
involvement depends upon the localization (peritone-
um vs rectum) and/or the phase (acute vs chronic) of
inflammation.
ACKNOWLEDGMENTS
The authors thank Colette Betouliere for helpful techni-
cal assistance, Dr. Million Mulugeta and Dr. R. Garcia-
Vilar for reviewingthe manuscript, and Institut National de
la Recherche Agronomique for financial support.
REFERENCES
1. Starr MS, West GB: Bradykinin and oedema formation in
heated paws of rats. Br J Pharmacol 31:178-187, 1967
2. Hargeaves KM, Troullos ES, Dionnes RA, Schmidt EA, Scha-
fer SC, Jorris JL: Bradykinin is increased during acute and

1920 Digestive Diseases and Sciences, Vol. 40, No. 9 (September 1995)
BRADYKININ AND VISCEROSENSITIVITY
chronic inflammation: therapeuthic implications. Clin Pharma-
col Ther 44:613-621, 1988
3. Barthon JM, Proud D: Bradykinin antagonists. Annu Rev
Toxicol 31 : 129-162, 1991
4. Regoli D, Barabc J: Pharmacology of kinins. Pharmaeol Rev
32:1-46, 1980
5. Steranka LR, Dehaas CJ, Vavrek RJ, Stewart JM, Enna SJ,
Snyder SH: Antinociceptive effects of bradykinin antagonists.
Eur J Pharmacol 136:261-262, 1987
6. Corrca CR, Calixto JB: Evidence for participation of B1 and
B2 kinin receptors in formalin-induccd response in the mouse.
BrJ Pharmacol 110:193-198, 1993
7. Whalley ET, Clegg S, Stcwart JM, Vavrek RJ: The effect of
kinin agonists and antagonists on the pain response of the
human blister base. Naunyn-Schmcidebergs Arch Pharmacol
336:652-655, 1987
8. Chau TT, Lewin AC. Walter TL, Carlson RP, Weichman BM:
Evidence for a role of bradykinin in experimental pain models.
Agents Actions 34:235-238, 1991
9. Bhoola KD, Figucra CD, Worthy K: Bioregulation of kinins:
Kallikreins, kininogens and kininases. Pharmacol Rev 44:1-44,
1992
10. Dray A. Perkins M: Bradykinin and inflammatory pain. Trends
Neurosci 16:99-104, 1993
11. Stcranka LR. Manning DC, Dehaas CJ, Ferkany JW, Borosky
SA, Ryder JR, Vavrek RJ, Stewart JM, Snyder SH: Bradykinin
as a pain mediator: Receptors are localized to sensory neurons
and antagonists have analgesic actions. Proc Natl Acad Sci
USA 85:3245-3252, 1988
12. Hock FJ, Wirth K, Albus U. Linz W, Gerhards H J, Wiemer G,
Henke St, Breipohl G, Konig W, Knolle J, Scholkens BA: Hoe
14(1 a new potent and long acting bradykinin antagonist; in
vitro studies. Br J Pharmacol 102:769-773, 1991
13. Wirth K, Hock FJ, Albus U, Linz W. Anagnostopoulos H,
Hcnke St, Breipohl G, Konig W, Knolle J, Scholkens BA: Hoe
140, a new potent and long acting bradykinin antagonist; in vivo
studies. Br J Pharmacol 102:774-777, 1991
14. Ness TJ, Gebhart GF: Colorectal distension as a noxious
visceral stimulus: Physiologic characterization of pseudo-
affective reflexes in the rat. Br Res 450:153-169, 1988
15. Morteau O, Hachet T, Caussette M, Bueno L: Experimental
colitis alters viseeromotor response to colorectal distension in
awake rats. Dig Dis Sci 39(61:1239-1248, 1994
16. Pairet M, Ruckebusch Y: On the relevance of non-steroidal
anti-inflammatory drugs in the prevention of paralytic ileus in
rodents. J Pharm Pharmacol 205:547-555, 1989
17. Ruckebusch Y, Fioramonti J: Electrical spiking activity and
propulsion in small intestine in fed and fasted rats. Gastroen-
terology 69:1045-1047, 1975
18. Heapy C, Shaw JS, Farmer SC: Differential sensitivity of an-
tinociceptive assays to the bradykinin antagonist Hoe 140. Br J
Pharmacol 108:209-213, 1993
Digestive Diseases and Sciences, Vol. 40. No. 9 (September 19951
19. Ferreira SH, Lorenzetti BB, Poole S: Bradykinin initiates cy-
tokine-mediatcd inflammatory hyperalgesia. Br J Pharmacol
110:1227-1231, 1993
20. Geppctti P, Maggi CA, Perretti F, Frilli F, Manzini S: Simul-
tancous release by bradykinin of substance P and CGRP like
immunoreactivities from capsaicin-sensitive structures in
guinea pig heart. Br J Pharmacol 94:288-290, 1988
21. Bertrand C, Geppetti P, Piedimonte G, Baker J, Petersson G,
Nadel JA: Bradykinin increases permeability in the rat nasal
mucosa: Contribution of angiotensin-converting enzyme and
neutral peptidase 24.11 Am Rev Respir Dis 154(4):A44, 1992
22. Julia V, Morteau O, Bueno L: Involvement of neurokinin 1 and
2 receptors in viseerosensitive response to rectal distension in
rats. Gastroenterology 107:94-102. 1994
23. Chapman V, Dickenson AH: The spinal and peripheral roles of
bradykinin and prostaglandins in nociceptive processing in the
rat. Eur J Pharmacol 219:427-433, 1992
24. Rangachari PK, Berezin M, Prior T: Effects of bradykinin on
the canine on the proximal colon. Regul Pept 46:511-522, 1993
25. Tien XY, Wallace I_J, Kachur JF, Won-Kim S, Gaginella TS:
Characterization of Ile, Ser-bradykinin-induced changes in
short-circuit current across rat colon. J Pharmacol Exp Ther
254(3):1063-1(167, 1990
26. Kuwahara A, Cooke HJ: Tachykinin-induced anion secretion
in guinea-pig distal colon: Role of neural and inflammatory
mediators. J Pharmacol Exp Ther 252:1-7, 1990
27. Davis AJ, Perkins MN: The involvement of bradykinin B1 and
B2 receptor mechanisms in cytokine-induced mechanical hy-
peralgesia in the rat. Br J Pharmacol 113:63-68, 1994
29. Tiffany CW, Bursch RM: Bradykinin stimulates tumor necrosis
factor and interleukin 1 release from macrophages. FEBS Lett
247:189-192, 1989
30. Birell G J, Mc Queen DS, Igoo A, Grubb BD: Prostanoid-
induced potentiation of the excitatory and sensitizing effects of
bradykinin on articular mechanoreceptors in the rat ankle
joint. Neuroscience 54(2):537-544, 1993
31. Kumazawa T, Mizumura K, Koda H: Involvement of EPs
subtype of prostaglandin E receptors in PGE2-induced en-
hancement of bradykinin response of nociceptors. Brain Res
632:321-324, 1993
32. Kumazawa T, Mizumura K, Koda H: Possible involvement of
the EP 2 receptor subtype in PGE2-induced enhancement of the
heat response of nociceptors. Neurosei Lett 175:71-73, 1994
33. Lerner UH, Brunius G, Anduren I, Berggren PO, Juntti-
Berggren L, Modeer: Bradykinin induces a B2 receptor-
mediated calcium signal linked to prostanoid formation in
human gingival fibroblasts in vitro. Agents Actions 37:43-52,
1992
34. Griesbacher T, Lembeck F: Effects of the bradykinin antago-
nist, Hoe 140, in experimental acute pancreatitis. Br J Phar-
macol 107:356-360, 1992
1921