Int Urol Nephrol (2007) 39:1055–1059
DOI 10.1007/s11255-007-9182-1
ORIGINAL PAPER
The effect of intravesical ketoprofen on acetylcholine-
evoked urinary bladder contractility and detrusor
overactivity in the anesthetized rabbit model
Eric Chieh-Lung Chou Æ Catherine Whitbeck Æ
Jeffrey Herz Æ Gregory A. Demopulos Æ
Robert M. Levin
Received: 12 December 2006 / Accepted: 12 January 2007 / Published online: 28 February 2007
Ó Springer Science+Business Media B.V. 2007
Abstract Intraurethral procedures such as the
transurethral resection of the prostate can gener-
ate detrusor overactivity and bladder irritability.
The rabbit model of detrusor overactivity has
proven to be an excellent model to study the
effects of drugs on detrusor overactivity. Using
this model, we evaluated the responses to intra-
vesical ketoprofen. In this model, each rabbit is
anesthetized and the right external carotid artery
is cannulated for blood pressure monitoring. A
catheter is inserted through the femoral artery and
This material is based upon work supported in part by
from grants from Omerous; in part by the Office of
Research and Development, Department of Veterans
Affairs, and in part by NIH grant RO-1-DK 067114.
E. C.-L. Chou
C. Whitbeck
R. M. Levin (&)
Albany College of Pharmacy, 106 New Scotland
Avenue, Albany, NY 12208, USA
e-mail: levinr@acp.edu
C. Whitbeck
R. M. Levin
Stratton VA Medical Center, Albany, NY, USA
E. C.-L. Chou
School of Medicine, China Medical College,
Taichung, Taiwan, ROC
E. C.-L. Chou
Department of Urology, China Medical College
Hospital Taichung, Taichung, Taiwan, ROC
J. Herz
G. A. Demopulos
Omerous Corporation, Seattle, WA, USA
is used for administration of acetylcholine (Ach).
The bladder is exposed and catheterized for
bladder pressure monitoring and drug addition
and the proximal urethra is ligated. Cystometry is
performed, the bladder drained, and 20 ml buffer
placed in the bladder. After 30 min Ach is injected
proximal to the vesical artery and the response of
the bladder and blood pressure is recorded. Ach
administration is repeated at 10-min intervals until
three consistent responses are obtained. The
bladder is drained and 20 ml of ketoprofen
(100 lM final concentration) is placed in the
bladder. Ach injections are repeated as given
above at 10 min intervals and observed for 4 h. At
the end of the experiment, a second cystometry is
performed. The following is a summary of the
results: Ketoprofen had no effect on either mic-
turition pressure or the intravesical volume at
micturition. Ketoprofen administration resulted in
a progressive 50% decrease in the response to
Ach. Ketoprofen mediated a progressive decrease
in detrusor overactivity amplitude and frequency,
reaching a maximum at 120–180 min.
Keywords Ketoprofen
Rabbits
Detrusor
overactivity
Acetylcholine
Introduction
One of the major problems with intraurethral
surgical procedures such as the transurethral
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1056
resection of the prostate (TURP) is the generation
of detrusor overactivity (unstable bladder con-
tractions), and the resultant bladder irritability,
frequency, urgency, and nocturia that follow this
operation [1–3]. These symptoms can remain for
several weeks following surgery. Treatment of the
detrusor overactivity by specific drugs would be
able to reduce the incidence and severity of
postsurgical detrusor overactivity. One such drug
which has proven to be effective in the treatment
of overactive bladder dysfunction is ketoprofen,
a relatively new orally aftive nonsteroidal anti-
inflammatory agent [4].
The rabbit model of detrusor overactivity has
proven to be an excellent model to study
the effects of drugs on unstable bladder contrac-
tions [5, 6]. Our experiments are based upon a
published technique in which each rabbit is
anesthetized and the carotid artery is cannulated
for blood pressure monitoring. A catheter was
inserted through the femoral until it is near the
vesical artery. This catheter is used for intra-
arterial administration of acetylcholine (Ach).
The bladder is exposed through a midline incision
and the bladder dome catheterized with a two-
channel 8 F catheter for both the monitoring of
bladder pressure and the administration of drugs
into the bladder. Approximately 50% of the
rabbits develop spontaneous detrusor overactivity
within approximately 30 min [7]. This model can
evaluate the effect of specific drugs on bladder
compliance, micturition pressure and capacity,
bladder instability, detrusor overactivity (ampli-
tude and frequency), and on the contractile
response to intra-arterial Ach.
Using this model, we evaluated the responses
to intravesical ketoprofen.
Methods
This study has been approved by the Institutional
Care and Use Committee of Stratton VA Medical
Centre, Albany, NY. Each mature New Zealand
white rabbit is anesthetized with an intramuscular
injection of ketamine/xylazine (25 mg ketamine,
10 mg xylazine/kg). Under anesthesia, the right
external carotid artery is cannulated for blood
pressure monitoring. A polyethylene catheter
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Int Urol Nephrol (2007) 39:1055–1059
(PE-90) is inserted through the rabbit’s right
femoral artery until it reaches the lower abdominal
aorta (approximately 0.5 cm above the bifurcation
of the aorta). A heparinized saline (30 IU ml–1)-
filled polyethylene catheter is used for intra-
arterial administration of Ach. To facilitate a
maximal dose of Ach getting access to the urinary
bladder, the rabbit’s left femoral artery is ligated.
The bladder is exposed through a midline
incision of the abdominal wall and the bladder
dome catheterized with an 8 F Foley catheter for
both the monitoring of bladder pressure and the
administration of test agents. The proximal ure-
thra is ligated to generate detrusor overactivity,
prevent micturition or leakage through the ure-
thra. The ureters are tied close to the bladder and
cut above the tie. The superpubic catheter is
connected via a three-way stopcock to an infusion
pump (Harvard Apparatus). One channel of the
stopcock is connected to a Statham pressure
transducer for monitoring bladder pressure.
Cystometry is performed with citric acid-sodium
citrate buffer (1 mM) using the infusion pump set
at an infusion rate of 1.40 ml/min. The bladder is
then emptied and filled with 20 ml with buffer.
Approximately 50% of these animal prepara-
tions develop detrusor overactivity within 30 min
of filling the bladder to 20 ml. After 30 min at
20 ml, 1 x 10–4 mole/kg of Ach is injected prox-
imal to the vesical artery and the response of the
bladder and blood pressure is recorded. Ach
administration is repeated three times at 10-min
intervals to ensure consistent responses.
After three consistent responses to Ach are
obtained, in the presence or absence of unstable
bladder contractions, the bladder is drained and
20 ml of ketoprofen (100 lM final concentration)
is placed in the bladder. Ach injections are
repeated as given above at 10 min intervals and
observed for 4 h. At the end of the experiment, a
second cystometry is performed.
Statistical significance was evaluated by analy-
sis of variance followed by a Neuman-Keuls test
for multiple comparisons.
Results
Figure 1 displays a representative example of a
control cystometric curve and micturition reflex
Int Urol Nephrol (2007) 39:1055–1059 1057

Cystometry Table 1 Data is given as percent of control

25
Intravesical Pressure

20
A Time (min) 0 (control) 30 60 120 180 240
(cm H2O)

15 Control BP 100 110 105 95 90 85

10 HR 100 105 110 98 94 86
5 Ketoprofen BP 100 114 108 100 97 83
HR 100 115 115 95 90 74
0
0 3 6 9 12 15 HR = heart rate = 98 +/– 14 beats per minute (control)
Time (minutes)
BP = blood pressure = 84 +/– 5 cm Hg (control)
Response to Intra-arterial Acetylcholine
120
B contractions remain fairly constant for each indi-
Intravesical Pressure Blood Pressure

100
(mm Hg)

80 vidual rabbit over the time course of the exper-

60 iment.
40 Table 1 presents the effects of ketoprofen on
25 blood pressure and heart rate. There was no
20 consistent or significant effect of ketoprofen on
(cm H2O)

15
blood pressure or heart rate. The minor decrease
10
observed at 240 min is due to the duration of
5
0
Ach Ach Ach anesthesia.
0 4 8 12 16 20 Figure 2 shows the effects of ketoprofen on
Time (minutes) cystometric values (micturition pressure and
Hyperreflexia bladder volume at micturition) performed prior
25
C to the drug administration and at the end of the
Intravesical Pressure

20
4 h study. Ketoprofen had no effect on either
(cm H2O)

15
parameter.
10
Figure 3 displays the effect of ketoprofen on
5
the contractile responses to intra-arterial Ach. In
0
0 3 6 9 12 15 the absence of drugs, the responses to Ach are
Time (minutes) consistent throughout the 4 h study. Ketoprofen
resulted in a progressive decrease in the response
Fig. 1 (A) Representative example of cystometric curve
and micturition reflex contraction. (B) Representative
to approximately 50 % of control.
example of three sequential intra-arterial injections of Figure 4 displays the effect of ketoprofen on
acetylcholine. (C) Representative example of detrusor detrusor overactivity amplitude and frequency.
overactivity (unstable bladder contractions)
25
Bladder Volume at Micturition (ml)

Pressure
(1A). From the cystometric curve, we calculate Capacity
Micturition Pressure (cm H2O)

20
the micturition pressure, functional bladder
capacity (volume at micturition), and compliance
15
(change in pressure per ml). Figure 1B displays a
representative responses to intra-arterial acetyl- 10
choline. It should be noted that Ach has effects on
both blood pressure and bladder pressure. These 5
responses can then be used to differentiate
between the cardiovascular and bladder effects 0
of ketoprofen. Figure 1C displays a representa- Pre-Drug Post-Drug
tive example of unstable bladder contractions. It
Fig. 2 Effects of ketoprofen on micturition pressure and
should be noted that, although the amplitude and
volume at micturition. Each bar is the mean +/– standard
frequency is different for individual rabbits, the error of the mean (SEM) of 4–6 rabbits. Asterisk indicates
amplitude and frequency of the unstable bladder significantly different from pre-drug

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1058 Int Urol Nephrol (2007) 39:1055–1059

140 Discussion
Response to Ach (% of Control)

120
This model of bladder instability is based on our
100 earlier studies in which penile ligatures (on
anesthetized rabbits) were used to generate
80 detrusor overactivity [5, 6]. In the current study
* * intra-abdominal ligation of the proximal urethra
60
* was used to generate the detrusor overactivity
*
* observed. This method produced a more consis-
40 Control
Ketaprofen tent detrusor overactivity in a larger percentage
20 of rabbits. Please note that in these studies, we
Control 30 60 120 180 240
Time after intravesical drug (min) use the terms unstable bladder contractions and
detrusor overactivity interchangeably.
Fig. 3 Effects of ketoprofen on the response to intra- Overactive bladder, the symptoms of fre-
arterial Ach. Each point is the mean + /– SEM of 4–6
quency, urgency, and urge incontinence are often
rabbits. Asterisk indicates significantly different from pre-
drug side effects of all transurethral procedures, and
can be quite severe [1, 2]. We believe that
urethral irritation, inflammation, and local dam-
140 age initiates the syndrome, in the same manner as
Hyperreflexia Amplitude
in our model, and thus drugs found useful in our
Amplitude of Hyperrreflexia

120
model should be translatable to use in man.
100 We chose ketoprofen to use in these initial
(% of Control)

studies because of its apparent effectiveness in

80
the treatment of unstable bladder contractions
60 * *
*
and detrusor overactivity [4, 9, 10]. In our studies,
intravesical ketoprofen had no significant effect
40 *
Control on cystometric values, which would indicate that
Ketaprofen
20 it would not interfere with micturition post
surgery, whereas it did significantly decrease the
140 phasic response to Ach and detrusor overactivity,
Hyperreflexia Frequency
thus being beneficial in the treatment of bladder
Frequency of Hyperrreflexia

120

100
instability. In regards to detrusor overactivity,
(% of Control)

ketoprofen displayed a consistent and effective

80
inhibition.
60 In conclusion, our model of detrusor overactiv-
40 *
ity is a useful model in evaluating the effectiveness
*
* * of intravesical drugs in the treatment of bladder
20 *
instability. The ability to quantify the effects on
0 blood pressure, micturition pressure and volume,
Control 30 60 120 180 240
and on the response to Ach are very useful in
Time after intravesical drug (min)
differentiating the effects of the drugs against
Fig. 4 Effects of ketoprofen on the amplitude and detrusor overactivity from effects on micturition,
frequency of bladder overactivity. Each point is the blood pressure, and muscarinic stimulation.
mean +/– SEM of 4–6 rabbits. Asterisk indicates signifi-
cantly different from pre-drug
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