Digestive Diseases and Sciences, VoL 39, No. 6 (June 1994), pp.

1239-1248

Experimental Colitis Alters Visceromotor

Response to Colorectal Distension
in Awake Rats
OLIVIER MORTEAU, MS, THIERRY HACHET, MS, MARCEL CAUSSETTE, BS,
and LIONEL BUENO, DSc

The influence o f intermittent colorectal distension (CRD) on proximal colonic motility and
abdominal pain perception was investigated in awake rats equipped with intraparietal
electrodes on the cecum, proximal colon, and abdomen, before and three days after
rectocolitis induction by trinitrobenzene sulfonic acid (TNB)/ethanol. The normal myo-
electrical activities o f cecum and proximal colon [5. 2 + 0.5 and 9. 7 +_O. 7 long spike bursts
(LSB) p e r 5 min, respectively] were significantly (P < O.05) and gradually decreased by
control CRD, at diameters above 9 mm. A t the maximum CRD diameter (13. 7 mm), 1.6
+-- 0.6 cecal and 3.9 + 0.8 colonic spike bursts occurred p e r 5 min (respectively, 69 and
60% decreases). This upstream inhibition was accompanied by a significant (P < O.05) and
gradual increase in abdominal contractions (0. 4 +_ O.4 p e r 5 min in control vs 23. 4 ++-1.9
in response to 13.7 m m in diameter). Three days after TNB/ethanol, visceromotor and
abdominal responses were significantly (P < O.05) enhanced at the least CRD diameter o f
9 m m (cecum: 3.1 +- O.4 after TNB vs 5. 0 +- O. 7 in control; proximal colon: 5.1 +- O.9 vs
9.3 +- 2.2; abdomen: 7. 7 +- 1.5 vs 0.5 +- 0.4). We conclude that in awake rats, CRD evokes
both abdominal contractions in response to pain and inhibition o f cecal and proximal
colonic motility, which thresholds are both lowered by TNB-induced rectocolitis.

KEY WORDS: rectum; distension; trinitrobenzene sulfonic acid; colitis; colon; motility; rat.

Inflammatory bowel diseases (ulcerative colitis,
Crohn's disease) are characterized by severe in-
flammation of the colonic mucosa (1, 2), associated
with strong colonic motor disturbances and acute
abdominal pain (3-5). Currently, both the mecha-
Manuscript received February 25, 1993; revised manuscript
received June 4, 1993; accepted June 30, 1993.
From the Department of Pharmacology, INRA, Toulouse,
France.
This work was presented at the Sixth European Symposium on
Gastrointestinal Motility, July 19-24, 1992, Barcelona, Spain,
and has appeared in abstract form in the Journal o f Gastrointes-
tinal Motility 4(3):233, 1992.
The authors thank INRA for its financial support.
Address for reprint requests: Dr. L. Bueno, Department of
Pharmacology INRA, 180 chemin de Tournefeuille, BP3, 31931
Toulouse Cedex, France.
nisms of colonic inflammation and the processes of
acute abdominal pain are studied independently by
the use of experimental animal models.
In rats, chronic colitis has been induced by intra-
colonic administration of trinitrobenzene sulfonic
acid (TNB) in ethanol (6-8). Intracolonic adminis-
tration of TNB/ethanol induces an acute inflamma-
tion with ulceration and infiltration of polymorpho-
nuclear lymphocytes and neutrophils and a eventual
chronic inflammatory response with luminal nar-
rowing and smooth muscle hypertrophy (6). This
colitis persists up to eight weeks (6) and is charac-
terized by high tissue myeloperoxidase activity and
decreased glutathione levels (9). Moreover, the me-
diators involved in TNB/ethanol-induced colitis are

Digestive Diseases and Sciences, VoL 39, No. 6 (June 1994) 1239
0163-2116/94/0600-1239507.00/0© 1994PlenumPublishingCorporation

similar to those implied in IBD, n a m e l y prostaglan-
din E2, t h r o m b o x a n e s A 2 and B2, leukotrienes B4
and D4, platelet-activating factor, and interleukin-1
(7, 10-16). T h e last two h a v e b e e n s h o w n to play a
role in colonic m o t o r alterations induced b y T N B /
ethanol in rats (8).
Several models of visceral nociception h a v e b e e n
d e v e l o p e d , b a s e d on the s t u d y o f v i s c e r o m o t o r ,
cardiovascular, or behavioral r e s p o n s e s to gastro-
intestinal distension in animals (17-23). T h e s e reli-
able models allow pharmacological evaluation of
drugs (18, 20, 21, 23-25) and physiological investi-
gation of neuronal m e c h a n i s m s implied in abdomi-
nal nociception (22). A m o n g these models, colorec-
tal distension (CRD) has b e e n d e v e l o p e d in rats to
evaluate behavioral, cardiovascular, and v i s c e r o -
m o t o r reflexes associated with pain (24).
A recent s t u d y gives e v i d e n c e that inflamed co-
Ionic m u c o s a leads to an increased a v e r s i v e b e h a v -
ior in r e s p o n s e to C R D in rats (26). H o w e v e r , no
e x p e r i m e n t a l w o r k has y e t a d d r e s s e d the changes in
colonic m o t o r r e s p o n s e to visceral noxious stimulus
in inflammatory conditions.
In this work, w e adapted electromyography to a
CRD model in awake rats, in order to investigate: (1)
the cecocolonic m o t o r reflex, (2) the m o t o r activity of
abdominal striated muscles, as a noxious response,
and (3) the possible alterations of these two responses
during TNB-induced rectocolonic inflammation.
M A T E R I A L S AND M E T H O D S
Animal Preparation. Male Wistar rats weighing 250-350
g were surgically prepared for electromyography, accord-
ing to a previously described technique (27). Rats were
anesthetized with 0.3 ml of acepromazine (0.5 mg/kg) and
0.3 ml of ketamine (Imalgene 1000, Rhrne-Mrrieux,
Lyon, France) injected intraperitoneally. After laparot-
omy, two groups of three electrodes of nichrome wire
were implanted in the cecal wall and two others in the
proximal colonic wall at 1 and 3 cm from the cecocolonic
junction. In addition, one group of three electrodes was
implanted in the abdominal striated musculature at 3 cm
from the locus of laparotomy. Electrodes were exterior-
ized on the back of the neck and protected by a glass tube
attached to the skin.
Motility Recordings. Electromyographic recordings be-
gan five days after surgery. The electrical activity of the
cecum and colon and of the abdominal striated muscles
was recorded with an electroencephalograph machine
(Mini VIII, Alvar, Paris, France) using a short time con-
stant (0.03 sec) to remove low-frequency signals (<3 Hz).
Colitis Induction. Trinitrobenzene sulfonic acid (TNB)
(80 mg/kg in 0.3 ml of ethanol 50%) was administered
intrarectally through a silicone catheter introduced at 1
cm from the anus.
1240
BUENO ET AL
Distension Protocol. Distensions were performed before
and three days after TNB/ethanoi administration. The rats
were placed in plastic tunnels, where they could move or
escape but not turn around, in order to prevent damage to
the balloon. They had been accustomed to this procedure in
order to prevent a stress reaction to the tunnel during
experiments. The balloon used for distension was an arterial
embolectomy catheter (Fogarty, Edwards Laboratories,
Inc., Santa Ana, California) having a 2 mm diameter and a
2 cm length (Figure 1A). It was rectally inserted in a mini-
mally invasive manner at 1 cm from the anus and fixed to
the tail. In order to control for possible effects due to
balloon insertion on motility, we waited until the recovery
of a normal myoelectrical activity before distension. The
balloon was then increasingly inflated in an intermittent
way, each step of inflation lasting 5 min (Figure 1B). The
choice of intermittent distension was based on clinical ob-
servations (28). The choice of a 5-min duration was based
on technical and ethical considerations. It was the minimum
duration required to evaluate eventual modification of the
cecocolonic motility on myoelectrical recordings while lim-
iting the pain duration for the animal. In order to detect an
eventual leakage, a syringe filled with water was used for
inflation and the volume withdrawn was controlled for each
distension. Between two increased steps of inflation, we
waited for a minimum of 5 min and until the motor activity
had returned to normal. The distension was interrupted
each time the rats exhibited a behavior that could indicate
excessive pain, like attempts to turn around or escape from
the tunnel, or intense vocalization. Balloons were calibrated
using an electronic caliper gauge, in order to define the
mean diameters corresponding to different volumes of infla-
tion (Figure 1C). The maximum pressure corresponding to
the maximum diameter of distension (13.7 mm) was evalu-
ated: it was 69.8 - 2.0 mm Hg. This maximum pressure was
in the range of the pressures generally used in visceral pain
studies and below to that commonly used (80 mm Hg) in the
previously described CRD model in rats (24).
According to the length of the colon (15-20 cm), the
closer group of colonic electrodes (3 cm from the ceco-
colonic junction) was located at 9-14 cm from the site of
distension (3 cm from the anus) (Figure 1D).
This protocol was approved by our local animal care
and use committee (certificate No. 91123, 22/03/91).
Measurement of Colonic Intraluminal Basal Pressures in
Response to Balloon Inflation Before and After TNB/
Ethanol Administration. A flaccid polyvinyl balloon (2 cm
length, 4 ml maximum air capacity) offering low resis-
tance to distension was built for pressure measurements.
The maximal air volume for which the resistance of the
balloon to inflation remained negligible was evaluated to
2.5 mi (Figure 2). The balloon was connected to an
electronic pressure transducer built in the laboratory
(Barostat, INRA, Toulouse, France). In order to estimate
the error due to air compression in the pressure trans-
ducer, we measured the compliance of the whole system
without the balloon (Figure 2). As the compliance curve
appeared to be linear and the variations of the volume
negligible with regard to the corresponding pressures, we
found no need to correct the values of the volume in-
jected. Six rats weighing 250-350 g were placed by turns
in a tunnel and the balloon was rectally inserted in a
Digestive Diseases and Sciences, Vot. 39, No. 6 (June 1994)
C O L O N I C MOTOR R E S P O N S E TO C O L O R E C T A L D I S T E N S I O N A N D I N F L A M M A T I O N

A
Arterial Embolectomy Catheters (Fogarty®)
........./ / .....

syringe filled with water balloon inflation hub balloon

2.0 ml
B

1.5 ml
ip,~m.lmq

IJ
1.0 ml I I
0.8 ml

0 °li
I I ......
! ! I
5 mln 5 mln

C D
14

12,

10
[ I
8.
|
| 6.

4.
2 cm

2' x -x- x electrodel ~ Jlcrn

~m
0 ~ ...................... (~
o.o 1.o l'.s .... b,,o,,
Volume (ml) U
Fig 1. Protocol of colorectal distension. A/ Balloon used for distension. 13/
Intermittent CRD protocol: increasing 5 min steps of distension (from 0.5 to 2.0
ml of water) alternate with 5 min steps of recuperation. CI Diameter/volume
relationship of the balloons. D/Position of electrodes and balloon in the intes-
tinal tract.

minimally invasive manner at 1 cm from the anus. The Statistical Analysis of Results. Statistical analysis of the
balloon was increasingly inflated with air in an intermit- curves was performed using computerized (personal com-
tent manner during 10 sec separated by 30 sec, up to 2.4 puter, XT 286, IBM) analysis of variance (ANOVA, pro-
ml of air injected, and the basal pressure response of the gram Mystat) followed b y an individual analysis of the
colorectal wall, expressed in millimeters of mercury was curve plots (Student's t test, paired values). Differences
read on the pressure transducer. In order to avoid exces- were considered significant for P < 0.05 and the values
sive pain due to an excessive increase in pressure, the expressed as mean -+ SEM.
pressure transducer was programmed to stop as soon as
the maximum value of 80 mm Hg was reached. Basal
values were taken as the lowest pressure values obtained
in response to one given volume of inflation. One series of RESULTS
five measurements was conducted daily in each rat for
four days running. TNB/ethanol was then intrarectally Basal Cecocolonic and Abdominal Myoelectrical
administered to the rats according to the described pro-
Activity. The myoelectrical activity of the cecum
tocol, m second series of pressure measurements was
performed under the same conditions on the treated rats and proximal colon was characterized by the pres-
from day 2 to day 5 after TNB/ethanol administration. e n c e o f s p i k e b u r s t s l a s t i n g 4.2 ___ 0.3 a n d 7.1 --- 0.4

Digestive Diseases and Sciences, Vol. 39, No. 6 (June 1994) 1241

60-
pressure transducer
55- D
50-
I balloon
45
A
O~
40
3:
E
s 35 f
8 30
w
25
D.
20
15
10
5
O_ ¢ ~,
0.0 0.5 1.0
Fig 2. Compliance curves of the balloon and of the pressure transducer used
to measure intracolonic pressures. Volumes are expressed in milliliters of air.
sec, respectively, and occurring at a mean rate of
5.2 --- 0.5 and 9.7 -.+ 0.7/5 min.
Abdominal striated muscles generally exhibited
no electrical signal in the absence of distension
stimulus. We just noticed from time to time (0.4 _+
0.4/5 min) the occurrence of short-lasting (3.3 -+- 0.3
sec) low-amplitude spike bursts (20, 50 I~V) due to
movements of the rat contracting its abdomen.
Visceromotor and Abdominal Responses to Con-
trol CRD. A significant influence of the CRD diam-
eter on cecocolonic motility and on the number of
abdominal contractions was observed, as given by
analysis of variance. The gradual increase in disten-
sion diameter induced a linear decrease in the num-
ber of cecocolonic spike bursts (Figure 4A), which
was significant (P < 0.05, N = 11) when a minimum
diameter of 10.8 mm was applied and remained
significant for higher diameters of distension, ie,
11.4, 12.7, and 13.7 mm (Table 1). The minimum
numbers of spike bursts (1.6 ± 0.6 and 3.9 +- 0.8/5
min for cecum and proximal colon, respectively)
were obtained for the maximum diameter of disten-
sion (13.7 mm) and corresponded to a 69 and 60%
inhibition for cecum and proximal colon, respec-
tively.
In response to CRD, we noted an increase in the
occurrence of spike bursts of the abdominal striated
muscles (Figure 4B). Like the cecocolonic re-
sponse, the abdominal response became significant
(P < 0.05) when the diameter of the rectal balloon
1242
BUENO ET AL
., , , , ,
1.5 2.0 2.5 3.0 3.5 4.0
Volume (ml)
reached 9 mm. For upper diameters, the increase in
abdominal spike bursts was positively related to the
increase in balloon diameter. The number of ab-
dominal spikes was 23.4 - 1.9/5 min at the maxi-
mum step of distension (diameter of the balloon:
13.7 mm) (Fig 4B, Table 1).
An example of the myoelectrical profiles of the
cecum, colon and abdominal musculature observed
in response to CRD is illustrated in Figure 3.
Visceromotor and Abdominal Responses to CRD
After TNB Treatment. Three days after rectal ad-
ministration of TNB/ethanol, no change was de-
tected in the myoelectrical activity of the cecum
and proximal colon, while the statistical (ANOVA)
influence of TNB/ethanol treatment was observed
on cecocolonic response to CRD. In response to
CRD after TNB/ethanol treatment, we noted a pro-
gressive decrease in the number of cecal and co-
lonic spike bursts (Figure 5A and B). However,
after rectocolitis induction, this decrease in the
number of cecal and colonic spike bursts was sig-
nificantly (P < 0.05) enhanced for a given diameter
of distension (Figure 5A and B, Table 1), Indeed, a
significantly (P < 0.05) lower number of colonic
spike bursts was observed for a diameter of 9 ram,
while no significant inhibition was detected before
TNB treatment. For a distension of 9 mm in diam-
eter before and after TNB/ethanol treatment, the
number of spike bursts per 5 min for cecum and
proximal colon were 3.1 -+ 0.4 vs 5.0 -+ 0.7 and 5.1
Digestive Diseases and Sciences, Vol. 39, No. 6 (June 1994)
COLONIC MOTOR RESPONSE TO COLORECTAL DISTENSION AND INFLAMMATION

Cecum , j-.-lOOlaV

0.0 ml Colon
r r 1 ! ! f F IOOpV
(~-- 2 mm) ...~ _,aL ~t .,.. ......... ~,. .., ~ kw ~-o
][ '~" 'l'r ~ "vW
Abdomen [-lO0~V
L 0

- [ t - ' t ° t° 'j v t - k It _ ,_o

0.5 ml
r T "f ; T ~ ! [-,oo.v
(~ = g mm) ,. k . & j..,t ,It . ~ ~. it t-.o
FlOO~tV
v

" ~ II l,~ , , ,~L_

L
~, ,
i I- ' ° ° " v
I t.-0
t, I vT , v ,1, ~,, ,,v ,y

0.8 ml 1,| : ~ J~i ,JI, ,, ~ F lO01av

( ~ = 11 ram) ;¢ 4 '~' l , v • ' -'It ,, ~ "L"O
-lOOpV
i,i ~4 , , , L L.,O

F100pV
.J.t 111 l.,tll=l l llJ.--a J lL
'.Tr TTI I'll IVY y V ITT, "- v TT JJ " ; I t..-o
T! "
1.5 ml J~ .* [-loo~,v
( ~ = 14 m m ) •. . , ,,, ,,, I :,v'" i Vl ,W.f i L.o
iJ JJl J [-loo~v
B ~ ~!V'.yw! r 11 wry y ,--o

Id I I 1 1 1 I I ~ J | I J ~l , , , lllllJ, d,dl F1001aV

Wll I l I I t 1 | I ~T y I vI ,1 ,11111 1, TlVVl I L~O

2.0 rnl ,,~.J L i i III J Ill I Jl . , lllJll, ,J4,ll F IOOpV

( ~ = 15 m m ) wq v v ~ I 11 I IT I I ~1 vl v T I v T I T T vl v n | L"O

~ i l J l l J t . t t d ~ l Jill,, dLd~ t-.ol-l°°~v

~yvvvryvT WIT v! ~TvyyTy, vTv~!
I * rain I
Fig 3. Responses of cecum and proximal colon, expressed in n u m b e r of spike bursts per 5 min,
to increasing diameters of distension, before (control) and after TNB/ethanol treatment. Signif-
icantly different (P -< 0.05) from values before CRD (diameter of 2 mm) (*) or from control values
(÷).

___ 0.9 v s 9.3 + 1.0, respectively. Similarly, at the
last step of distension (diameter: 13.7 mm), the
n u m b e r of spike bursts for c e c u m and proximal
colon w e r e 1.2 ± 0.4 v s 1.6 ± 0.6 and 1.9 ± 0.5 v s
3.9 ± 0.8 before and after T N B / e t h a n o l t r e a t m e n t ,
respectively. At this diameter, values w e r e not sig-
nificantly different (P < 0.05) f r o m controls (Figure
5A and B).
T h r e e d a y s after rectal administration of T N B /
ethanol, no significant change w a s detected on the
basal n u m b e r of abdominal contractions, while the
statistical ( A N O V A ) influence of the t r e a t m e n t w a s

Digestive Diseases and Sciences, Vol. 39, No, 6 (June 1994) 1243
 BUENO ET AL

TABLE 1. NUMBER OF CECAL, COLONIC, AND ABDOMINALSPIKE BURSTS PER 5 MIN BEFORE AND AFTER TNB/ETHANOL TREATMENT
IN RESPONSE TO VARIOUS DIAMETERS OF DISTENSION*

Control CRD CRD after TNB treatment

Diameter
(mm) Cecum Proximal colon Abdomen Cecum Proximal colon Abdomen
2.0 5.2 -+ 0.5 9.7 +-- 0.7 0.4 __- 0.4 4.7 +-- 0.5 8.5 --- 0.9 0.9 --- 2.4
9.0 5.0 -+ 0.7 9.3 - 1.0 0.5 -+ 0.4 3.1 _+ 0.4b 5.1 -+ 0.9b 7.7 -+ 1.5b
10.8 4.0 +-- 0.5at 6.6 -+ 0.7a 9.3 -+ 2.2a 2.6 -+ 0.7a 4.1 -4- 0.9a 12.7 -+ 1.4a
11.4 3.5 -+ 0.5a 5.5 -+ 1.1a 13.1 -+ 2.3a 2.2 --- 0.7a 3.4 -+ 0.7a 14.2 --- 1.5a
12.7 2.7 +- 0.Sa 4.5 -+ 1.1a 19.7 -+ 2.3a 1.5 -+ 0.4a 2.4 _+ 1.0a 15.1 --- 1.5a
13.7 1.6 -+ 0.6a 3.9 -+ 0.8a 23.4 +- 1.9a 1.2 +-- 0.4a 1.9 _ 0.5a 17.4 +- 1.3a

*Data given as mean _+ SEM ( N = 11).

t a , significantly different (P <- 0.05) from values before CRD (diameter of 2 mm) or b, from control values.

observed on the abdominal response to CRD. After
TNB/ethanol administration, a significant (P <
0.05) enhancement of the increase in abdominal
spike bursts in response to CRD was noted for a
lower diameter of distension (9 mm): 7.7 - 1.5 vs
0.5 -+ 0.4 in control CRD (Figure 5C). For upper
diameters of distension, the occurrence of abdomi-
nal contractions was no more significant when com-
pared to control values.
Intracolonic Basal Pressure Measurements Before
and After TNB/Ethanoi Treatment. In control con-
ditions~ the six rats exhibited reproducible basal
pressure responses to increasing volumes of infla-
tion. No significant intraindividual nor interindivid-
ual variations were observed during the four days of
control experiments (Figure 6A). At days 2, 3, 4,
and 5 after TNB/ethanol administration, the six rats
exhibited characteristic symptoms of previously de-
scribed (6) colorectal inflammation (weight loss,
diarrhea), confirmed by macroscopic examination
after killing (day 5). The pressure response of each
rat did not vary significantly during the test period
(two to five days after TNB/ethanol administration).
None of the animals exhibited a statistically in-
creased basal pressure response to balloon inflation
(P > 0.05), when compared to control experiments
(Figure 6A and B). However, the amplitude of the
colorectal contractions was enhanced after colitis
induction and the highest measurable cutoff value of
80 mm Hg was often reached before the final infla-
tion step (2.4 ml). Figure 6C illustrates the mean
basal pressure response of the six rats before and
after colitis induction. No statistical difference (P >
0.05) was detected after point by point comparison
of the two curves.
Behavioral Observations. Rats often stayed still in
the tunnels during distension. Minor changes in
behavior were observed for the highest values of
CRD: washing and moderate writhing. In two rats,
1244
which showed intense writhing, the experiment was
interrupted.
DISCUSSION
In this work, we provide evidence that in rats
distension of the rectum and of the terminal part of
the colon inhibits the motor activity of the proximal
colon by reducing the number of spike bursts. This
response is very reproducible with low intra- or
interindividual variations and is correlated to the
diameter of distension. An ascending intestinointes-
tinal inhibitory reflex to distension had been de-
scribed previously in both animals (17) and humans
(29) at jejunal and gastroduodenal levels. In rats,
behavioral, cardiovascular, and visceromotor re-
sponses (defined as contractions of abdominal and
hindlimb musculature and a relaxation of the anal
sphincters) to colorectal distension have been stud-
ied (24). However, to our knowledge, no previous
study performed in rats reported a visceromotor
reflex similar to the one we describe here. In hu-
mans, a motor response to rectal distension has
been observed (28), but it was limited to an increase
in rectal pressure occurring at the site of mechanical
stimulation. In animals, distension of the proximal
colon of anesthetized rats induces an increase in
proximal colonic pressure at the same locus (20).
Due to the distance separating the site of disten-
sion from that of recordings, ie, 9-14 cm, the inhib-
itory response we observe here is taken as a long
intestinointestinal inhibitory reflex. In dogs, Frantz-
ides et al (30) demonstrated an intrinsic neural path-
w a y for the long intestinointestinal inhibitory reflex.
However, the stimulus they used was arterial ad-
ministration of neostigmine, which involves imme-
diate contractions locally. In all the studies con-
cerning long-distance inhibition of intestinal
motility by a distending balloon (29, 31-33), the
Digestive Diseases and Sciences, Vol. 39, No. 6 (June 1994)
COLONIC MOTOR RESPONSE TO COLORECTAL DISTENSION AND INFLAMMATION

A 30

control ~
._~ 25 *
E
E
gm 4 20 **
JO *
15 ,
Q,
m
"~ 2 10 "~
iE .= T
E
Z
z= 5

4
I
6
1 I
8 10
I
12
I 1
14 0 ....... 1°°°i*°'
0 2 4 6 8 10 12 14

12_ B

~,~¢ontrol
Diameter (ram)

~_~
Fig Diameter (mm)
5. Intracolonic pressure response to increasing volumes of
inflation. (A) Pressure/volume relationship of six rats in control
conditions. Each curve is the mean representation of 4 days of
A 10_ measurements. (B) Pressure-volume relationship of the same
animals after TNB/ethanol administration. Each curve is the
g 8-
mean representation of measurements at days 2, 3, 4, and 5 after
TNB/ethanol treatment. (C) Mean pressure-volume relationship

TNBlet~
II
of the rats before (control) and after TNB/ethanol administra-
+ * • tion.
o
6-

4_ ered as a somatic event related to pain. Abdominal

! contractions may consist of involuntary attempts to
E expel the rectal balloon as a cause of visceral dis-
z 2-
0 I I I I I
0 4 6 8 10 12
Diameter (ram)
Fig 4. Typical alteration of cecal, colonic, and abdominal elec-
trical activity in response to increasing diameters of distension,
observed in a fasted rat in control conditions. The spike bursts of
short duration observable on cecal and colonic recordings are
artifactual signals of an abdominal nature.
neural pathway involved is considered to be the
extrinsic sympathetic pathway with either a prever-
tebral ganglionic or spinal relay, rather than an
intrinsic nervous pathway. In view of these data, it
could be suggested that the inhibitory visceromotor
reflex we observe here is extrinsically mediated.
Moreover, in our study, the inhibitory reflex on the
colon was always accompanied by a stimulatory
effect on the abdomen. This perfect match of the
responses supports the involvement of extrinsic
nervous system in the colonic inhibitory reflex we
observe.
The contractile response of the abdominal wall
we identify by electromyography may be consid-
comfort or pain. However, these abdominal con-
tractions are also observed in other abdominal pain
I models such as intraperitoneal acetic acid adminis-
14
tration (34), suggesting that they correspond to in-
voluntary movements associated with pain. Fur-
thermore, these abdominal contractions have
previously been described as accompanying other
symptoms of visceral pain such as "tonic increases
in respiration, heart rate, and blood pressure" (35),
and electromyographic recordings of abdominal
contractions have been used previously as a crite-
rion of pain in response to CRD (24). Consequently,
the abdominal response appears to be related to
pain perception, and the number of movements may
serve as an index of pain intensity. The threshold
diameter required to induce significant abdominal
motor activity may occur at a point at which ab-
dominal discomfort gives rise to acute pain.
The use of TNB/ethanol to induce chronic inflam-
mation of the rectocolonic mucosa was developed
by Morris et al (6), who observed sites of inflam-
mation extending from the perirectal region to 7 cm
from the anus. While the presence of inflammation
in the proximal colon is associated with altered

Digestive Diseases and Sciences, Vol. 39, No. 6 (June 1994) 1245
 BUENO ET AL

": ) ,.'
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Fig 6. Response of the abdominal striated muscles, expressed in number of spike
bursts per 5 min, to increasing diameters of distension, before (control) and after
TNB/ethanol treatment. Significantly different (P _< 0.05) from values before
CRD (diameter of 2 ram) (*) or from control values (+).

myoelectrical activity (8), no motor change in the
proximal colon was observed in the present study,
confirming that inflammation was limited to the
distal colon (6). Our results show that, after recto-
colitis induction by TNB/ethanol, colonic motor
inhibition and abdominal cramps occur for a lower
diameter of distension than in control rats. Our
observation agrees with a recent behavioral study in
rats where abdominal pain evoked by CRD was
increased after the colorectal mucosa had been in-
flamed with turpentine (26). Furthermore, a very
recent work gives evidence that in cats the distal
esophagus inflamed with turpentine is more sensi-
tive to distension than a noninflamed one (36). In
our study, the important point is that experimental
colitis induces both colonic and abdominal re-
sponses at a diameter of distension that normally
induces no response. This fact clearly leads to the
conclusion that recal inflammation turns normal co-
Ionic perception into an abnormal one.
Since it has been reported that rats treated with
TNB/ethanol develop some areas of bowel thicken-
ing (6), the lowering of the colonic inhibitory and
abdominal response threshold we observed after
TNB administration may be attributed to a decrease
in the diameter of the colonic lumen. However, we
detected no significant change in the basal pressure
response of the inflamed colonic wall to increasing
volumes of distension when compared to normal
colonic wall. Consequently, we exclude the hypoth-
esis that alteration of the visceromotor and abdom-
inal responses evoked by colitis is due to a purely
mechanical phenomenon. Among the hypotheses to
explain the hypersensitivity, we retain the possibil-
ity of the release of mediators involved in visceral
pain, motor alterations, and inflammation, such as

1246 Digestive Diseases and Sciences, Vot. 39, No. 6 (June 1994)
COLONIC MOTOR RESPONSE TO COLORECTAL DISTENSION AND INFLAMMATION
serotoninergic compounds, tachykinins, or brady-
kinin. Another hypothesis is that experimental rec-
tocolitis may involve changes in nervous afferent
endings at the enteric level, as has been shown in
the IBD-induced loss of some mucosal neuropep-
tidic innervation (37).
From a clinical point of view, rectal distension
has been shown to induce increased anorectal sen-
sitivity in patients with ulcerative colitis when com-
pared to normal subjects (38). Moreover, our obser-
vation that in rats inflamed rectocolonic mucosa
may give rise to inhibition of the proximal colon is
in agreement with the fact that in some patients with
active proctocolitis, fecal matter accumulates in the
uninflamed colon, above an area of active colitis
(39-42), and that proximal colonic transit is slowed
in patients with ulcerative colitis (43).
In conclusion, we developed a reproducible
model of colorectal distension in rats, which may
allow pharmacological evaluation of the mecha-
nisms mediating abdominal pain and colonic motor
alterations in response to a distending stimulus.
This model may also provide valuable information
on the mechanism by which colonic mucosal in-
flammation alters viscerosensitivity in some patho-
physiological conditions.
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