Cardiovascular Research 67 (2005) 647 – 654

www.elsevier.com/locate/cardiores

Endothelin receptor—A blockade decreases ventricular arrhythmias after

myocardial infarction in rats
Giannis G. Baltogiannisa, Dimitrios G. Tsalikakisb, Agathokleia C. Mitsia,
Konstantinos E. Hatzistergosc, Dimitrios Elaiopoulosa, Dimitrios I. Fotiadisb,
Zenon S. Kyriakidesd, Theofilos M. Kolettisa,*
a
Department of Cardiology, University of Ioannina, 1 University Avenue, 45110 Ioannina, Greece
b
Department of Computer Sciences, University of Ioannina, 1 University Avenue, 45110 Ioannina, Greece
c
Department of Pathology, University of Ioannina, 1 University Avenue, 45110 Ioannina, Greece
d
Department of Cardiology, Hellenic Red Cross Hospital, 27 Venizelou Street, 17123 Athens, Greece

Received 5 February 2005; received in revised form 17 April 2005; accepted 20 April 2005
Available online 23 May 2005
Time for primary review 17 days

Abstract

Objective: Endothelin-1 (ET-1) production increases during acute myocardial infarction (MI) and may contribute to the genesis of
ventricular tachycardia (VT) and ventricular fibrillation (VF). However, the antiarrhythmic effects of ET-1 receptor blockade, examined
shortly after MI, have been debated. In the present study, we examined the effects of such treatment on VT/VF during the first 24 h post-MI.
Methods: Thirty-five Wistar rats (223 T 22 g) were randomly allocated to either the ET-1 receptor-A (ETA) antagonist BQ-123 (0.4 mg/kg,
BQ-123 group, n = 17), or normal saline (control group, n = 18) and were subjected to coronary artery ligation. A single-lead
electrocardiogram was continuously recorded for 24 h post-MI, using an implanted telemetry system, and episodes of VT/VF were
analyzed. Monophasic action potential (MAP) recordings were obtained from the left (LV) and right (RV) ventricular epicardium at baseline,
5 min after treatment and 24 h post-MI.
Results: There were 15.94 T 19.35 episodes/h/rat of VT/VF in the control group and 1.66 T 2.22 in the BQ-123 group ( p = 0.010), resulting in
a lower ( p = 0.030) arrhythmic mortality in treated animals. The mean episode duration was 7.40 T 7.16 s for the control group and 2.30 T 1.37
s for the BQ-123 group ( p = 0.011). The maximum decrease in VT/VF was observed during the 1st, 5th and 6th hours post-MI. In the control
group, LV MAP duration increased 24 h post-MI, displaying an increased beat-to-beat variation, but remained unchanged in the BQ-123
group.
Conclusion: Acute ETA blockade reduces the incidence of VT/V F during the first 24-h post-MI in the rat, through a decrease in the
dispersion of repolarization.
D 2005 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.

Keywords: Arrhythmia (mechanisms); Endothelins; Infarction

1. Introduction one-third of patients with MI die shortly after acute coronary

Acute myocardial infarction (MI) remains a common
cause of morbidity and mortality worldwide [1]. Early
reperfusion strategies decrease mortality, but approximately
* Corresponding author. Tel.: +30 265 1097533; fax: +30 265 1097017.
E-mail address: thkolet@cc.uoi.gr (T.M. Kolettis).
0008-6363/$ - see front matter D 2005 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.
doi:10.1016/j.cardiores.2005.04.020
occlusion, before receiving medical attention [1,2]. The vast
majority of these deaths are caused by polymorphic
ventricular tachycardia (VT) and ventricular fibrillation
(VF) [1,2].
In the constellation of mechanisms leading to ischaemia-
induced ventricular arrhythmias, endothelin-1 (ET-1) may
play a significant role. Experimental [3] and clinical [4,5]
studies have shown that the production of ET-1 increases
648 G.G. Baltogiannis et al. / Cardiovascular Research 67 (2005) 647 – 654
markedly during MI, but the pathophysiological signifi-
cance of this finding is not well understood. In addition to
its vasoconstrictive, pro-fibrotic and pro-inflammatory
actions, contributing to myocardial necrosis, [6,7] ET-1
has been implicated in the pathogenesis of VT and VF
during the acute phase of MI [3 – 5]. Based on these
observations, it has been hypothesized that blockade of
ET-1 receptors during MI may confer antiarrhythmic
actions. Previous studies examining this hypothesis pro-
duced conflicting results, [8 – 14] but the wide variation in
the experimental protocols utilized precludes firm conclu-
sions. Moreover, all previously published studies limited the
time window for the observation of ventricular arrhythmias
up to 2 h post-MI induction.
In the present study, we explored further the pathophys-
iological role of ET-1 during MI. We aimed to examine the
effects of acute ET-1 receptor blockade (a) on the infarct
size and (b) on ventricular arrhythmias and to provide
further insight into the possible electrophysiologic mechan-
isms of such intervention. To clarify the differences in the
previously reported results, we chose, first, to examine the
effects of selective ET-1 receptor-A (ETA) blockade
administered acutely at the maximum dosage known to
produce local myocardial pharmacologic action and second,
to avoid the confounding effects of reperfusion, since the
mechanisms of reperfusion-induced arrhythmias are likely
to differ from those responsible for ischemic arrhythmias
[15]. The animal model used in the present study was the rat
model, which offers clear-cut advantages in the study of
ventricular arrhythmias. The rat not only exhibits a high
frequency of VT and VF in response to myocardial
ischaemia, but also the time course of their occurrence
corresponds to the arrhythmia time course seen in humans
after acute MI [16,17]. In the present study, we extended the
previously studied time window and observed the total
arrhythmia burden during the first 24 h post-MI.
2. Methods
The study was conducted in 35 female Wistar rats, 20 T 2
weeks old, weighing 200 –250 g. The animals received
appropriate care and the investigation conforms to the Guide
for the Care and Use of Laboratory Animals published by
the US National Institutes of Health (NIH Publication No.
85-23, revised 1996). All rats were housed in individual
cages in a climate-controlled environment, with a 12 : 12-
h light– dark cycle and were given water and standard rat
chow ad libitum.
2.1. Implantation of telemetry transmitter
A continuous electrocardiogram (ECG) telemetry trans-
mitter (Dataquest, Data Sciences International, Transoma
Medical, Arden Hills, MN, USA) was implanted in the
abdominal cavity, using a previously described method
[16]. Under ether anaesthesia, the animals were intubated
and mechanically ventilated using a rodent ventilator
(model 7025, Ugo Basile, Comerio, VA, Italy). Anaesthe-
sia was maintained with a mixture of oxygen and 2%
isoflurane. The transmitter was secured in the abdominal
cavity, and the leads were tunnelled under the skin and
attached to the underlying tissue. The positive electrode
was placed in a V4 – V5 position and the negative electrode
was placed under the right axilla. All incisions were
sutured in two layers and the rat was then housed in an
individual cage, placed on a receiver that continuously
captured the signal, independently of animal activity. The
ECG signal was displayed in real-time with the use of a
computer program (A.R.T. 2.2, Dataquest, Data Sciences
International, Transoma Medical, Arden Hills, MN, USA).
After confirmation of a good quality ECG signal, the data
were recorded continuously for the following 48 h and
stored for analysis.
2.2. Monophasic action potential recordings
Twenty-four hours after the telemetry transmitter
implantation, the rats were re-anesthetized and a left
thoracotomy was performed, allowing dissection of the
pectoral muscles. The heart was exposed and the pericar-
dium was carefully removed. Monophasic action potential
(MAP) epicardial signals were recorded, as described
previously [18]. A MAP probe (model 200, EP Technol-
ogies, Sunnyvale, CA, USA) was placed on the lateral left
(LV) and right (RV) ventricular walls, exerting mild,
constant pressure against the epicardium to eliminate
electrical artifacts. The signal was amplified with the use
of a preamplifier (model 300, EP Technologies, Sunnyvale,
CA, USA) and filtered at 50 Hz, using a digital notch
filter. A continuous data stream was fed into a personal
computer equipped with an analog-to-digital converter
(BNC 2110, National Instruments Corporation, Dallas,
TX, USA) and 2-min recordings were obtained, as soon as
a clear, steady state signal was achieved. The recording
software utilized in this study has been developed at our
Institution, using Labview 6.1. This software has been
validated with the use of a pulse generator, capable of
producing a sequence of square waves with 1 mV
amplitude, as described previously [19]. Furthermore, this
software program facilitates the analysis of MAP record-
ings and details of these features are presented elsewhere
[19]. For purposes of this study, maximal signal amplitude
and action potential duration at 90% of repolarization
(APD90) were measured. These measurements were
performed as follows: at baseline, 5 min after drug
administration and 24 h after MI. During analysis of the
MAP recordings, non-sinus beats were excluded and 50
consecutive sinus beats per recording were analyzed. The
standard deviation of APD90 was calculated for each
recording, as a measure of beat-to-beat variation, indicating
electrical alternans [18,20,21].
 G.G. Baltogiannis et al. / Cardiovascular Research 67 (2005) 647 – 654
2.3. Drug administration and generation of MI
Following baseline MAP recordings, the rats were
randomized in 1 : 1 fashion to receive either normal saline
or BQ-123, the most selective ETA peptide antagonist [22]
(Cyclo (d-Asp-Pro-d-Val-Leu-d-Trp) I Na, C31H41N6O7 I
Na, molecular weight 632.7, EMD Biosciences, Inc, Merck
KGaA, Darmstadt, Germany). The dosage used was 0.4 mg/
kg, which corresponds to that previously found to produce
maximal pharmacological effects locally in the myocardi-
um, but without any significant systemic effects [8,23 – 25].
In order to achieve a fast onset of a pharmacologic action, a
slow (within 3 min) infusion, directly in the LV cavity was
performed [26]. Minor bleeding, occasionally seen at the
puncture site, stopped after light pressure was applied
locally.
Five minutes after drug administration, coronary artery
ligation was performed, as described previously [27]. In
brief, the heart was exteriorized from the thorax and the left
coronary artery was encircled and ligated using a 6-0 suture,
placed between the pulmonary artery cone and the left atrial
appendage. The heart was returned to its normal position
and the thorax was closed in two layers. The remaining air
was aspirated from the thorax, allowing the rats to resume
spontaneous respiration. A six-lead ECG was obtained and
ST elevation in 2 or more leads was considered a proof of
induced MI.
The animals were returned to their cage and ECG
recording was continued for another 24 h, or until
spontaneous death. Bradyarrhythmic death occurring
during the first 5 min after induction of MI was attributed
to the surgical procedure or to cardiogenic shock [16] and
these animals were excluded from the study. No
resuscitation attempts were allowed at any time during
the study.
Twenty-four hours after the generation of MI, the
survivors were re-anaesthetized and the site of previous
left thoracotomy was reopened. The heart was exposed
and MAP recordings were repeated at the same sites.
The rats were then sacrificed using a lethal dose of
potassium chloride and the heart was harvested for
measurement of infarct size. The study protocol is
depicted in Fig. 1.
2.4. Infarct size
The method used for measurement of infarct size has
been described previously [28]. In brief, the heart was
excised, frozen (in 20 -C for 1 h) and hand-cut
simultaneously in five 2 mm slices. They were incubated
(in triphenyltetrazolium chloride for 15 min at 37 -C) and
fixed (in 10% formalin for 20 min). The slices were scanned
with the use of a high resolution scanner (Scanjet 4570c/
5500c, Hewlett-Packard, Palo Alto, CA, USA). The areas of
the infarcted and non-infarcted myocardium were deter-
mined by planimetry, using a previously validated software
649
Fig. 1. Study protocol.
program (Image Tool, University of Texas, USA). Infarct
size, expressed as a percentage of the LV cross sectional
area, was defined as the ratio between the infarcted area,
divided by the total LV area.
2.5. Heart rate
Because heart rate (HR) in the conscious rat can be
variable, we analyzed continuous 5-min ECG recordings,
from which non-sinus beats were excluded. The mean value
of these RR intervals was used to determine HR. HR was
calculated at baseline, at the 5th, 30th and 60th minute post-
MI and hourly thereafter.
2.6. Arrhythmia analysis
The acquired single-lead ECG tracings were displayed
and analyzed off-line independently by two of the authors
(G.G.B., D.E.). The stored tracings were manually scrolled
on a computer screen and the observers recorded all
arrhythmic events. Ventricular arrhythmias were recorded
as single ventricular ectopic beats (VEB’s), couplets,
triplets, VT and VF, according to the guidelines provided
by the Lambeth Conventions for determination of experi-
mental arrhythmias [29]. Even with these guidelines,
separating VF from VT was often difficult, and this has
been the experience of others [16]. Therefore, in this study,
we report the sum of VT and VF episodes. The duration of
each VT and VF episode was measured using the time-scale
provided by the recording software. To account for the
censoring effect associated with differential survival, the
arrhythmia frequencies are reported as mean values per hour
per rat.
2.7. Statistical analysis
All values are given as mean T one standard deviation,
unless otherwise specified. Mortality rates were compared
using Yates’ corrected chi square. Differences in continuous
650 G.G. Baltogiannis et al. / Cardiovascular Research 67 (2005) 647 – 654
variables were compared using Student’s t-test, or the
analysis of variance for repeated measures, followed by
Tukey’s multiple comparisons test, as appropriate. The
continuous variables describing the arrhythmia frequencies
were not normally distributed and were compared using the
Mann –Whitney U-test. Statistical significance was defined
at an alpha level of 0.05.
3. Results
We studied female 40 Wistar rats, weighing 224 T 21 g.
Two rats (one had received BQ-123 and one normal
saline) died during the surgical procedure and were
excluded from the study. Three further rats (two had
received BQ-123 and one normal saline) died within the
first 5 min of complete atrioventricular block (probably
attributable to cardiogenic shock) and were also excluded.
Thirty five animals were included in the study, of which
17 rats (223 T 21 g) were randomized to receive BQ-123
(BQ-123 group) and 18 rats (224 T 22 g) to receive normal
saline (control group).
3.1. Mortality
During the 24-h period following MI, none of the
animals in the BQ-123 group (0 / 17, 0%) died due to
tachyarrhythmias, whereas 6 animals (6 / 18, 33%) in the
control group had a fatal episode of VF ( p = 0.030). Two
rats in the BQ-123 group (2 / 17, 11%) and one rat in the
control group (1 / 18, 5%) died due to bradycardia associated
with complete atrioventricular block. The overall mortality
did not differ significantly between the two groups
( p = 0.14).
3.2. Heart rate
HR was not significantly different between the two
groups during the entire 24-h observational period
( F = 1.34, p = 0.22, Fig. 2).
Fig. 2. Sinus heart rate in the two groups. No significant differences were
found.
Fig. 3. Distribution of ventricular tachycardia (VT) and ventricular
fibrillation (VF) expressed as mean (T standard error of the mean) duration
(number of episodes times duration of each episode) per hour post-
myocardial infarction. Shaded area indicates the presumed time-period of
maximum drug effect.
3.3. Infarct size
Infarct size was calculated for the 26 survivors, 15 in the
BQ-123 group and 11 in the control group. Mean infarct
size was 39.7 T 4.2% for the BQ-123 group and 38.6 T 5.0%
for the control group ( p = 0.56).
3.4. Ventricular arrhythmias
No statistically significant differences were found be-
tween the two groups in the number of VEBs, couplets or
triplets. There was a significant reduction in the incidence of
VT + VF episodes, from 15.94 T 19.35 episodes/h/rat in the
control group to 1.66 T 2.22 episodes/h/rat in the BQ-123
group ( p = 0.010). Furthermore, there was a significant
decrease in the mean duration of each episode, from
7.40 T 7.16 s in the control group to 2.30 T 1.37 s in the
BQ-123 group ( p = 0.011). The hourly distribution of total
VT + VF duration (number of episodes times duration of
each episode) is shown in Fig. 3. Significant differences
between the two groups were observed during the first
( p = 0.0033), fifth ( p = 0.0085) and sixth ( p = 0.017) hours
post MI. This timing reflects the time period of maximum
treatment action in conjunction with the highest incidence of
ventricular arrhythmias.
3.5. Monophasic action potential duration and amplitude
No changes in RV APD90 were found in either group
( F = 0.26, p = 0.77). There was a significant variance in LV
APD90 ( F = 71.7, p < 0.0001), that was due to a significant
increase in LV APD90 24 h post-MI in the control group,
compared to baseline (Table 1). In contrast, no significant
changes in LV APD90 were observed over time in the BQ-
123 group (Table 1, Fig. 4A and B). Furthermore, a
significant variance was found in beat-to-beat variability
of LV APD90 ( F = 34.0 p < 0.001). This variance was due to
a significant increase in beat-to-beat variability of LV
APD90 24-h post-MI in the control group, compared to
 G.G. Baltogiannis et al. / Cardiovascular Research 67 (2005) 647 – 654 651

Table 1 complicated with heart failure [4]. Animal studies suggest

Changes in amplitude and duration of action potential and beat-to-beat that ET-1 may contribute to the induction of VT and VF
variability in action potential duration over time in the two groups
independently of ischaemia, [8– 14,30,31] but these direct
BQ-123 Control
electrophysiologic actions of ET- 1 are still debated [8,13].
APD90. baseline (ms) 107 T 7 106 T 7 This issue is further complicated by the fact that conclusions
APD90. after injection (ms) 106 T 5 106 T 6
have been drawn from exogenously administered ET-1 in
APD90. 24-h post-MI (ms) 109 T 5 131 T10*
some studies [31] and from the actions of endogenous ET-1
in others [9,10].
Beat-to-beat variability‘ baseline (ms) 2.8 T 0.6 3.7 T 0.6 In the present study, we hypothesized that acute
Beat-to-beat variability‘ after injection (ms) 3.1 T 0.9 3.8 T 1.5 administration of BQ-123, an ETA antagonist, may exert
Beat-to-beat variability‘ 24-h post-MI (ms) 4.7 T 1.9 14.9 T 5.6*
antiarrhythmic effects, by inhibiting the arrhythmogenic
potential of endogenous ET-1. We tested this hypothesis in a
Amplitude baseline (mV) 7.7 T 0.4 7.8 T 0.4 rat model of MI; the rat exhibits a high frequency of
Amplitude after injection (mV) 7.7 T 0.4 7.8 T 0.4 ischaemic ventricular arrhythmias in a repetitive, self-
Amplitude 24-h post-MI (mV) 3.8 T 0.7* 3.5 T 0.4*
Data are given as mean T standard deviation.
* p < 0.05 compared to baseline.
. APD90: action potential duration at 90% of repolarization.
‘
Expressed as the standard deviation of APD90 in 50 consecutive sinus
beats.

baseline ( p = 0.0001). These findings were associated with

the occurrence of early afterdepolarizations (Fig. 4C). In
contrast, no significant changes in beat-to-beat variability
were observed over time in the BQ-123 group. LV MAP
amplitude decreased significantly 24 h post-MI in both
groups, with no significant differences between them. All
values are shown in Table 1.

4. Discussion

Sudden cardiac death accounts for approximately 50% of

the estimated 500,000 cardiovascular deaths that occur
annually in the United States, and similar figures apply in
Europe [2]. Acute MI is often responsible for sudden death
in patients without a prior history of heart disease, in whom
a fatal ventricular arrhythmia may be the first manifestation
of coronary atherosclerosis.
The pathophysiology of ischaemia-induced ventricular
arrhythmias is complex, and several aspects remain obscure.
Within seconds of coronary occlusion, metabolic and ionic
changes occur locally in the ischaemic ventricular myocar-
dium, resulting in dispersion of conduction and refractori-
ness, which, in turn, favour the onset of reentrant ventricular
arrhythmias [17].
ET-1 is a 21-amino acid peptide, acting on two specific
receptors, namely ETA and ETB, located in the vasculature
and cardiac muscle. Among other disease states, ET-1 levels
are elevated in acute coronary syndromes [3 –5]. Experi-
mental studies have shown increased ET-1 expression in the
infarcted area from 1 h up to 7 days after coronary occlusion
[3]. Similarly, elevated plasma concentrations of ET-1 have Fig. 4. Representative examples of monophasic action potential recordings
from the left ventricular epicardium in a rat at baseline (A), 24 h post-
been reported in patients with acute MI and have been
myocardial infarction in a rat in the BQ-123 group (B) and in the control
shown to predict 1-year mortality [4,5]. This rise peaks 6 group (C). Note the increase in duration and the beat-to-beat variability in
h after coronary artery occlusion and returns to normal the duration of the signal in C but not in B. Arrows indicate early after
values 24 h later, but it can be prolonged in infarcts depolarizations.
652 G.G. Baltogiannis et al. / Cardiovascular Research 67 (2005) 647 – 654
terminating manner, rendering this model ideal in the
assessment of various therapeutic interventions [16]. Fur-
thermore, the miniature telemetry recording system used in
our study facilitates the study of arrhythmias for extended
periods of time in the conscious rat, without the confound-
ing effects of anaesthesia [16].
We report a significant reduction in the incidence of
summed VT and VF episodes after acute ETA blockade and
a significantly shorter duration of each of these episodes.
This reduction in the total arrhythmic burden decreased
arrhythmic mortality. As expected from the comparable
infarct size, mortality due to cardiogenic shock was similar
in the two groups, resulting in a modest, non-significant
decrease in total mortality.
These findings cannot be explained by a decrease in the
necrotic myocardial mass, as no significant difference in
the infarct size was found in our study, and this is in
accordance with previously reported results [13,32].
Similarly, the antiarrhythmic effect observed in the present
report cannot be attributed to a lesser sympathetic
stimulation in treated rats, as suggested by studies
evaluating the effects of chronic ET-1 receptor blockade
post-MI [33]. This view is based on the absence of
significant differences in heart rate between the control
group and the BQ-123 treated group found in our study, as
well as in previous studies of acute ET-1 receptor blockade
post-MI [15,32].
The reduction in the incidence of VT and VF episodes
could be attributed to the haemodynamic effects of ETA
blockade. Indeed, a significant reduction in LV end-diastolic
pressure was reported after administration of a dual ET-1
receptor antagonist early post-MI [32]. In the present study,
we did not perform haemodynamic measurements; hence,
such a mechanism cannot be excluded. However, we feel
that this is unlikely, because the dose of BQ-123 used in this
study was carefully selected to produce effective ETA
blockade locally in the myocardium, without any systemic
effects [23 – 25]. Furthermore, Sharif et al. [8] using a total
dose of BQ-123 identical to ours, reported no effects on
blood pressure in the rat model of MI.
Two possible mechanisms may explain our findings.
First, ETA blockade may exert direct antiarrhythmic
effects, independently of ischaemia [30,31]. ET-1 activates
potassium currents and inhibits L-type calcium currents, an
effect mediated by ETA stimulation [34,35]. Furthermore,
infusion of ET-1 in dogs has been shown to induce
sustained polymorphic VT and VF [31]. Thus, acute
blockade of ETA, prior to MI induction, may ameliorate
the detrimental effects of the early surge in ET-1 that
occurs in the rat model [3] and in humans after MI [4,5].
Our results, in accordance with previous observations, [8–
10] point towards a direct antiarrhythmic effect of ETA
blockade post-MI. This antiarrhythmic effect of BQ-123
occurs within a very narrow dose range, [14] which may,
in part, explain the negative results reported previously
[11 – 13]. Other potential explanations include species
differences, variations in the anaesthesia and ischaemia–
reperfusion protocols, as well as in the route of adminis-
tration and nature of ET-1 receptor antagonists utilized [8–
14]. Of note, BQ-123 may exert a direct electrophysiologic
effect unrelated to its action at the ETA receptor. This agent
has unique antifibrillatory properties among ETA receptor
antagonists, as previously indicated from findings in
isolated rabbit ventricular cardiomyocytes, [36] and in
Langendorff-perfused rat hearts [37].
Second, the decreased arrhythmogenicity in the BQ-123
group may be due to a reduction of ischaemia within or
around the infarct zone. Previous work has indicated a
cardioprotective effect of ETA blockade, an effect mediated
by increased local release of nitric oxide [38]. In accordance
with these observations, we report a significant prolongation
of LV APD90 in the control group, indicative of myocardial
ischaemia, [17,18] which was eliminated in BQ-123-treated
rats. This prolongation may generate early afterdepolariza-
tions that are thought to be involved in the genesis of ET-1-
induced arrhythmias [30]. Furthermore, MAP duration in
the control group displayed a significant beat-to-beat
variation 24 h post-MI, an effect abolished by BQ-123.
This alternans in MAP duration is typical of ischaemia and
has been linked to the development of ischemic tachyar-
rhythmias [18,20,21]. Thus, a decreased dispersion of
ventricular repolarization, either via a direct antiarrhythmic
action of BQ-123, or secondary to a reduction in ischaemia,
may explain the antiarrhythmic effect of ETA blockade seen
in our study.
4.1. Limitations of the study
We feel that our study adds significantly to the current
understanding of the pathophysiological role of ET-1
during acute MI. However, apart from the absence of
haemodynamic data, discussed earlier, a few more limita-
tions may be apparent. First, our sample size was
relatively small, thus our study may be underpowered to
detect significant differences in total mortality. Second, we
did not assess the effects of acute ETA blockade on indices
of LV remodelling, which may correlate with the occur-
rence of ventricular arrhythmias. However, we believe that
measurement of such indices as early as 24 h post-MI
would be of lesser physiological significance. Third,
measurements of MAP recordings at some point during
the course of MI, e.g. 1 h post-MI induction, would have
permitted better assessment of the underlying pathophys-
iologic mechanisms of ET-1 receptor blockade. However,
these measurements were omitted, because they would
have interfered with the arrhythmia recording process.
Similarly, measurements of the ventricular effective refrac-
tory period would have added to the strengths of our study,
although no significant differences were found from our
group in a previous study in humans with coronary artery
disease [39]. Lastly, we did not address a long-standing
controversial issue, namely whether selective receptor-A,
 G.G. Baltogiannis et al. / Cardiovascular Research 67 (2005) 647 – 654
or mixed ETA and ETB blockade would result in better
effects post-MI.
In conclusion, acute administration of the selective ETA
antagonist BQ-123, in the rat model of acute MI, did not
affect infarct size, but decreased the incidence of malignant
ventricular arrhythmias during the first 24 h post-MI. In
keeping with previous suggestions, [35] this effect appears
to be the result of a reduction in dispersion of repolarization
across the ventricular myocardium. Whether this effect is
secondary to a decrease in ischaemia or whether it
represents a direct antiarrhythmic effect remains to be seen.
The clinical implications of these findings may be signif-
icant, as they provide further insight into the pathophysio-
logical mechanisms of sudden cardiac death. Additional
studies are needed to determine whether these favourable
results are sustained in the presence of reperfusion and to
assess the effects of chronic ET-1 receptor blockade.
References
[1] Rouleau JL, Talajic M, Sussex B, Potvin L, Warnica W, Davies R, et al.
Myocardial infarction patients in the 1990s—their risk factors,
stratification and survival in Canada: the Canadian Assessment of
Myocardial Infarction (CAMI) study. J Am Coll Cardiol 1996;27:
1119 – 27.
[2] Myerburg RJ, Kessler KM, Castellanos A. Sudden cardiac death:
structure, function, and time-dependence of risk. Circulation 1992;
85(suppl I):I2 – 10.
[3] Loennechen JP, Stoylen A, Beisvag V, Wisloff U, Ellingsen O.
Regional expression of endothelin-1, ANP, IGF-1, and LV wall stress
in the infarcted rat heart. Am J Physiol 2001;280:H2902 – 10.
[4] Miyauchi T, Yanagisawa M, Tomizawa T. Increased plasma concen-
trations of endothelin-1 and big endothelin-1 in acute myocardial
infarction. Lancet 1989;2:53 – 4.
[5] Omland T, Lie RT, Aakvaag A, Aarsland T, Dickstein K. Plasma
endothelin determination as a prognostic indicator of 1-year mortality
after acute myocardial infarction. Circulation 1994;89:1573 – 9.
[6] Ramires FJ, Nunes VL, Fernandes F, Mady C, Ramires JA.
Endothelins and myocardial fibrosis. J Card Fail 2003;9:232 – 7.
[7] Watanabe T, Suzuki N, Shimamoto N, Fujino M, Imada A.
Contribution of endogenous endothelin to the extension of myocardial
infarct size in rats. Circ Res 1991;69:370 – 7.
[8] Sharif I, Kane KA, Wainwright CL. Endothelin and ischemic
arrhythmias—antiarrhythmic or arrhythmogenic? Cardiovasc Res
1998;39:625 – 32.
[9] Raschack M, Juchelka F, Rozek-Schaefer G. The endothelin-A
antagonist LU135252 suppresses ischemic ventricular extrasystoles
and fibrillation in pigs and prevents hypoxic cellular decoupling. J
Cardiovasc Pharmacol 1998;31:145 – 8.
[10] Alberola Aguilar AM, Revert F, Moya A, Beltran J, Garcia J, San
Martin E, et al. Intravenous BQ-123 and phosphoramidon reduce
ventricular ectopic beats and myocardial infarct size in dogs
submitted to coronary occlusion and reperfusion. Gen Pharmacol
2000;35:143 – 7.
[11] Vago H, Soos P, Zima E, Geller L, Kekesi V, Andrasi T, et al. The ET-
A receptor antagonist LU135252 has no electrophysiological or anti-
arrhythmic effects during myocardial ischaemia/reperfusion in dogs.
Clin Sci (Lond) 2002;103(Suppl 48):223S – 7S.
[12] Richard V, Kaeffer N, Hogie M, Tron C, Blanc T, Thuillez C. Role of
endogenous endothelin in myocardial and coronary endothelial injury
and ischaemia and reperfusion in rats: studies with bosentan, a mixed
ETA – ETB antagonist. Br J Pharmacol 1994;113:869 – 76.
653
[13] Szabo T, Geller L, Merkely B, Selmeci L, Juhasz-Nagy A, Solti F.
Investigating the dual nature of endothelin-1: ischaemia or direct
arrhythmogenic effect? Life Sci 2000;66:2527 – 41.
[14] Garjani A, Wainwright CL, Zeitlin IJ, Wilson C, Slee SJ. Effects of
endothelin-1 and the ETA-receptor antagonist, BQ123, on ischemic
arrhythmias in anesthetized rats. J Cardiovasc Pharmacol 1995;25:
634 – 42.
[15] Curtis MJ, Hearse DJ. Ischaemia-induced and reperfusion-induced
arrhythmias differ in their sensitivity to potassium: implications for
mechanisms of initiation and maintenance of ventricular fibrillation.
J Mol Cell Cardiol 1989;21:21 – 40.
[16] Opitz CF, Mitchell GF, Pfeffer MA, Pfeffer JM. Arrhythmias and
death after coronary artery occlusion in the rat. Circulation 1995;92:
253 – 61.
[17] Horacek T, Neumann M, Mutius S, Budden M, Meesmann W.
Nonhomogeneous epicardial changes and the bimodal distribution of
early ventricular arrhythmias during acute coronary artery occlusion.
Basic Res Cardiol 1984;79:649 – 67.
[18] Franz MR. Current status of monophasic action potential recording:
theories, measurements and interpretations. Cardiovasc Res 1999;41:
25 – 40.
[19] Tsalikakis DG, Fotiadis DI, Kolettis T, Michalis LK. Automated
system for the analysis of heart monophasic action potentials. Comput
Cardiol 2003;30:339 – 42.
[20] Mohabir R, Franz MR, Clusin WT. In vivo electrophysiological
detection of myocardial ischaemia through monophasic action
potential recording. Prog Cardiovasc Dis 1991;34:15 – 28.
[21] Downar E, Janse MJ, Durrer D. The effect of acute coronary artery
occlusion on subepicardial transmembrane potentials in the intact
porcine heart. Circulation 1977;56:217 – 24.
[22] Davenport AP, Battistini B. Classification of endothelin receptors
and antagonists in clinical development. Clin Sci (Lond) 2002;
103(Suppl 48):1S – 3S.
[23] Spratt JC, Goddard J, Patel N, Strachan FE, Rankin AJ, Webb DJ.
Systemic ETA receptor antagonism with BQ-123 blocks ET-1 induced
forearm vasoconstriction and decreases peripheral vascular resistance
in healthy men. Br J Pharmacol 2001;134:648 – 54.
[24] Haynes WG, Webb DJ. Contribution of endogenous generation of
endothelin-1 to basal vascular tone. Lancet 1994;344:852 – 4.
[25] Kyriakides ZS, Kremastinos DT, Kolettis TM, Tasouli A, Antoniadis
A, Webb DJ. Acute endothelin-A receptor antagonism prevents
normal reduction of myocardial ischaemia on repeated balloon
inflations during angioplasty. Circulation 2000;102:1937 – 43.
[26] Harada K, Kawahara J, Okada Y, Uzumaki H, Kusaka M, Tokiwa T.
Effects of KRN4884 (a novel K+ channel opener) levromakalin,
nilvadipine and propanolol on endothelin-1-induced heart disorders in
anesthetized rats. Jpn J Pharmacol 1998;78:261 – 8.
[27] Pfeffer MA, Pfeffer JM, Fishbein MC, Fletcher PJ, Spadaro J, Kloner
RA, et al. Myocardial infarct size and ventricular function in rats. Circ
Res 1979;44:503 – 12.
[28] Ytrehus K, Liu Y, Tsuchida A, Miura T, Liu GS, Yang X, et al. Rat and
rabbit heart infarction: effects of anaesthesia, perfusate, risk zone, and
method of infarct sizing. Am J Physiol 1994;267:H2383 – 90.
[29] Walker MJ, Curtis MJ, Hearse DJ, Campbell RWF, Janse MJ, Yellon
DM, et al. The Lambeth Conventions: guidelines for the study of
arrhythmias in ischaemia, infarction, and reperfusion. Cardiovasc Res
1988;22:447 – 55.
[30] Yorikane R, Koike H, Miyake S. Electrophysiological effects of
endothelin-1 on canine myocardial cells. J Cardiovasc Pharmacol
1991;17(Suppl 7):S159 – 62.
[31] Szokodi I, Horkay F, Merkely B, Solti F, Geller L, Kiss P, et al.
Intraepicardial infusion of endothelin-1 induces ventricular arrhyth-
mias in dogs. Cardiovasc Res 1998;38:356 – 64.
[32] Clozel M, Qiu C, Qiu C-S, Hess P, Clozel J-P. Short-term endothelin
receptor blockade with tezosentan has both immediate and long-term
beneficial effects in rats with myocardial infarction. J Am Coll Cardiol
2002;39:142 – 7.
654 G.G. Baltogiannis et al. / Cardiovascular Research 67 (2005) 647 – 654
[33] Mulder P, Richard V, Derumeaux G, Hogie M, Henry JP, Lallemand
F, et al. Role of endogenous endothelin in chronic heart failure.
Effect of long-term treatment with an endothelin antagonist on
survival, hemodynamics and cardiac remodeling. Circulation 1997;
96:1976 – 82.
[34] Ono K, Tsujimoto G, Sakamoto A, Eto K, Masaki T, Ozaki Y, et al.
Endothelin-A receptor mediates cardiac inhibition by regulating
calcium and potassium currents. Nature 1994;370:301 – 4.
[35] Duru F, Barton M, Lüscher TF, Candinas R. Endothelin and cardiac
arrhythmias: do endothelin antagonists have a therapeutic potential as
antiarrhythmic drugs? Cardiovasc Res 2001;49:272 – 80.
[36] Kelso EJ, Spiers JP, McDermott BJ, Scholfield CN, Silke B.
Stimulation of L-type Ca2+ current by the endothelin receptor A-
selective antagonist, BQ-123 in ventricular cardiomyocytes isolat-
ed from the rabbit myocardium. Biochem Pharmacol 1998;55:
897 – 902.
[37] Crockett TR, Scott GA, McGowan NW, Kane KA, Wainwright CL.
Anti-arrhythmic and electrophysiological effects of the endothelin
receptor antagonists, BQ-123 and PD161721. Eur J Pharmacol 2001;
432:71 – 7.
[38] Gourine AV, Gonon AT, Pernow J. Involvement of nitric oxide in
cardioprotective effect of endothelin receptor antagonist during
ischaemia – reperfusion. Am J Physiol Heart Circ Physiol 2001;280:
H1105 – 12.
[39] Kolettis TM, Kyriakides ZS, Leftheriotis D, Papalambrou A,
Kremastinos DT, Webb DJ. Electrophysiologic effects of endothelin
receptor-A blockade in patients with coronary artery disease. J Interv
Card Electrophysiol 2003;8:173 – 9.