Am J Physiol Heart Circ Physiol 300: H2288H2299, 2011.
First published April 1, 2011; doi:10.1152/ajpheart.00140.2011.
Effect of dietary omega-3 fatty acids on the heart rate and the heart rate
variability responses to myocardial ischemia or submaximal exercise
George E. Billman1,2 and William S. Harris3
1
Department of Physiology and Cell Biology, 2Davis Heart and Lung Research Institute, Ohio State University, Columbus,
Ohio, and 3Sanford Research Institute, University of South Dakota, Sioux Falls, South Dakota
Submitted 9 February 2011; accepted in final form 24 March 2011
Billman GE, Harris WS. Effect of dietary omega-3 fatty acids
on the heart rate and the heart rate variability responses to myo-
cardial ischemia or submaximal exercise. Am J Physiol Heart Circ
Physiol 300: H2288 H2299, 2011. First published April 1, 2011;
doi:10.1152/ajpheart.00140.2011.The consumption of omega-3
polyunsaturated fatty acids (n-3 PUFAs) has been reported to decrease
resting heart rate (HR) and increase heart rate variability (HRV).
However, the effects of n-3 PUFAs on these variables in response to
a physiological stress (e.g., exercise or acute myocardial ischemia),
particularly in postmyocardial infarction (MI) patients, are unknown.
Therefore, HR and HRV (high frequency and total R-R interval
variability) were evaluated at rest, during submaximal exercise, and
during a 2-min coronary artery occlusion at rest and before and 3 mo
after n-3 PUFA treatment in dogs with healed MI (n 59). The dogs
were randomly assigned to either placebo (1 g/day corn oil, n 19)
or n-3 PUFA supplement (docosahexaenoic acid eicosapentaenoic
acid ethyl esters; 1 g/day, n 6; 2 g/day, n 12; or 4 g/day, n 22)
groups. The treatment elicited significant (P 0.01) dose-dependent
increases in right atrial n-3 PUFA levels but dose-independent reduc-
tions in resting HR and increases in resting HRV. In contrast, n-3
PUFAs did not attenuate the large changes in HR or HRV induced by
either the coronary occlusion or submaximal exercise. These data
demonstrate that dietary n-3 PUFA decreased resting (i.e., preexercise
or preocclusion) HR and increased resting HRV but did not alter the
cardiac response to physiologic challenges.
parasympathetic nervous system; exercise; fish oil; docosahexaenoic
acid; eicosapentaenoic acid
ALTERATIONS IN CARDIAC AUTONOMIC regulation have been impli-
cated in an increased risk for sudden cardiac death (5). In
particular, reduced cardiac parasympathetic activity coupled
with an enhanced sympathetic activation in both patients and
animals following myocardial infarction is associated with an
increased incidence of malignant ventricular tachyarrhythmias
and sudden cardiac death (5). Therefore, considerable effort
has been expended in the search for therapeutic interventions
that improve cardiac balance and could thereby protect against
sudden death in high-risk patient populations.
To be effective, these interventions must not only improve
baseline or resting cardiac autonomic balance but also maintain
these changes when the heart is challenged by a physiologic
stress such as acute myocardial ischemia or exercise. For
example, low doses of cholinergic muscarinic antagonists (at-
ropine or scopolamine) have, paradoxically, been shown to
enhance resting cardiac vagal regulation in both humans (14,
19, 32) and animals (23, 29). It was therefore proposed that the
continuous administration of low doses of muscarinic antago-
Address for reprint requests and other correspondence: G. E. Billman, Dept.
of Physiology and Cell Biology, Ohio State Univ., 304 Hamilton Hall, 1645
Neil Ave., Columbus, OH 43210-1218 (e-mail: billman.1@osu.edu).
H2288 0363-6135/11 Copyright 2011 the American Physiological Society
nists could improve cardiac vagal regulation and, as a conse-
quence, would also reduce the incidence of sudden cardiac
death in myocardial infarction patients (14, 19). However,
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these interventions failed to prevent ventricular fibrillation
induced by myocardial ischemia (23, 29) despite significant
increases in resting time and frequency domain markers of
heart rate variability (HRV; indexes of cardiac parasympathetic
regulation; Refs. 2, 48). It was subsequently shown that low
doses of atropine did not alter the cardiac autonomic response
elicited by either submaximal exercise or acute myocardial
ischemia (23). Thus the search for effective interventions that
improve autonomic balance both at rest and during stress
condition remains elusive.
A number of experimental and clinical studies report that
dietary omega-3 polyunsaturated fatty acids (n-3 PUFAs; Ref.
16) or the acute intravenous administration (9) of these lipids
can both lower resting heart rate (HR) and increase resting
HRV, data consistent with an enhanced baseline cardiac para-
sympathetic tone. It has been proposed that the cardiovascular
benefits ascribed to dietary n-3 PUFAs could result, at least in
part, from these reductions in HR and the corresponding
putative improvements in cardiac autonomic balance (16).
However, this concept has been challenged (11, 22, 24) and, to
date, the effects of dietary n-3 PUFA on HR and HRV during
physiological stress have received little attention. In a similar
manner, several lines of evidence suggest that the modulation
of cardiac autonomic function may not be solely responsible
for the n-3 PUFA-induced reductions in HR. For example, the
acute application of n-3 PUFA reduced the contraction rate of
neonatal rat myocyte cultures (31) while dietary n-3 PUFA
supplements decreased HR of isolated rat atria (33) and cardiac
transplant patients (25). A reduced pacemaker current (If)
density has also been recorded in sinus nodal myocytes from
rabbits fed n-3 PUFAs compared with cells from the hearts of
animals given similar amounts of sunflower oil (50). However,
whether these changes in intrinsic pacemaker rate are main-
tained during physiological stress also remains to be deter-
mined.
It was, therefore, the purpose of the present study to
evaluate the effects of dietary n-3 PUFAs (1 4 g/day for 3
mo) on the HR and the HRV responses to physiological
stressors [exercise (submaximal exercise, assessed at its
onset and at its termination) and acute myocardial ischemia
(2-min coronary artery occlusion)] in dogs with healed
myocardial infarctions. In particular, the hypothesis that the
dietary n-3 PUFA-mediated reductions in HR and increases
in HRV would be maintained during a physiologic challenge
was tested.
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 n-3 FATTY ACIDS INCREASE HRV AT REST BUT NOT DURING METABOLIC STRESS
METHODS
All the animal procedures were approved by the Ohio State Uni-
versity Institutional Animal Care and Use Committee and conformed
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).
Surgical Preparation
A timeline for the studies may be found in Fig. 1. Eighty heart-
worm free mixed breed dogs (2- to 3-yr-old male, n 21, and female,
n 59) weighing 19.3 0.2 kg (range: 13.6 25.9 kg) that were part
of an ongoing investigation of the cardiovascular effects of dietary n-3
PUFA (11) were used in this study. The animals were anesthetized
and instrumented to measure a ventricular electrogram (from which
HR and HRV were subsequently determined) and left circumflex
coronary artery blood flow as previously described (3, 11). A hydrau-
lic vascular occluder (model OC3; In Vivo Metric, Healdsburg,
CA) was placed around the left circumflex coronary artery and
used to induce acute myocardial ischemia for the coronary occlu-
sion experiments described below (see HRV Protocols). The left
anterior descending coronary artery was also isolated during the
instrumentation surgery, and a two-stage occlusion of this artery
was then performed approximately one-third the distance from its
origin to produce an anterior wall myocardial infarction (16% of
left ventricular mass; Ref. 3). This vessel was partially occluded
for 20 min and then tied off. Similar instrumentation surgery was
performed on six additional sham infarction dogs (i.e., the left
anterior descending artery was isolated but not ligated). The dogs
were given analgesic, antibiotic, and anti-arrhythmic therapy to
alleviate postoperative pain, to prevent postoperative infection, and
to reduce acute arrhythmias associated with the myocardial infarc-
tion as has been previously described (3, 11).
HRV Protocols
HRV was calculated using a Delta-Biometrics vagal tone monitor
triggering off the ECG R-R interval (Urbana, Champaign, IL). This
device employs the time-series signal processing techniques as devel-
oped by Porges to estimate the amplitude of respiratory sinus arrhyth-
mia [the high frequency (HF) component of R-R interval variability;
Ref. 42]. Details of this analysis have been described previously (6,
28). Data were averaged over 30-s intervals either during exercise or
the coronary occlusion. The following indexes of HRV were deter-
mined: vagal tone index, the HF (0.24 to 1.04 Hz) component of R-R
interval variability, and standard deviation (SD) of the R-R intervals
(a marker of total variability) for the same 30-s time periods.
The studies began 3 4 wk after the production of the myocardial
infarction. Fifteen (18.8%, 6 males, 9 females) died within 72 h of the
myocardial infarction, and reliable ECG recordings could not be
obtained in six dogs (1 male and 5 females). Thus HRV studies were
completed on 59 (14 male and 45 female) dogs following the myo-
cardial infarction and all six sham-operated dogs. First, over the
period of 35 days, the dogs learned to run on a motor-driven
treadmill. The cardiac response to submaximal (i.e., 60 70% of
maximal HR) exercise was then evaluated as follows: exercise lasted
a total of 18 min with workload increasing every 3 min. The protocol
began with a 3-min warm-up period, during which the dogs ran at
Fig. 1. Study timeline. Pre- and posttreatment data collection studies consist of
the following: three submaximal exercise studies and one 2-min coronary
artery occlusion at rest. Studies were performed at least 24 h apart. MI,
myocardial infarction.
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4.8 kph at 0% grade. The speed was then increased to 6.4 kph, and the
grade was increased every 3 min (0, 4, 8, 12, and 16%). The
submaximal exercise test was repeated three times (1/day). On a
subsequent day, with the dogs lying quietly unrestrained on a table, a
2-min left circumflex coronary occlusion was made. Left circumflex
blood flow, HR, and HRV were monitored continuously throughout
the exercise or occlusion studies. The submaximal exercise and the
coronary occlusion at rest studies were performed both before and
after 3 mo of treatment with either placebo (1 g/day corn oil) or daily
n-3 PUFA capsules (1 4 g/day).
Dietary Omega-3 Polyunsaturated Fatty Acid Protocol
The dogs were place on a diet that did not contain any n-3 PUFAs
(Harlan Teklad; Harlan Laboratories, Indianapolis, IN) beginning 1
wk before the instrumentation surgery and were maintained on this
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diet until the end of the study (4 mo). After the pretreatment data
collection (3 4 wk after the surgery) had been collected, the dogs
were then randomly assigned to the following groups: placebo (n
19, 3 male and 16 female); 1 g/day n-3 PUFA (n 6, all female); 2
g/day n-3 PUFA (n 12, 7 male and 5 female); 4 g/day n-3 PUFA
(n 22, 4 male and 18 female); and sham 4 g/day sham (n 6, 1
male and 5 females). The dogs were given supplements similar to
those used in the GISSI-Prevenzione study (20). The n-3 PUFA group
received 465 mg ethyl eicosapentaenoate (EPA) 375 mg ethyl
docosahexaenoate (DHA) (375 per capsule; Lovaza; GlaxoSmithK-
line, Research Triangle Park, NC). The placebo was corn oil (1 g, 58%
linoleic acid 28% oleic acid). The capsules were given per os before
the daily feeding (between 8:00 and 10:00 AM each day, 7 days per
wk for 3 mo).
Red Blood Cell and Cardiac Tissue Fatty Acid Analysis
Fasting blood samples (5 ml) were drawn into EDTA tubes from a
cephalic vein between 8:00 and 9:00 AM 1 day before the initiation of
the treatment (placebo or n-3 PUFA) and when tissue was harvested
at the end of the study (14 wk after the treatment began). Right atrial
and left ventricular tissues were obtained when the hearts were
harvested; the tissue and red blood cells (RBC) were flash frozen in
liquid nitrogen and stored at 80C for future analysis.
RBC and phospholipids from cardiac tissue were analyzed for fatty
acid composition using previously described techniques (12, 35). The
samples were analyzed by gas chromatography using a GC2010-FID
(Shimadzu, Columbia, MD) equipped with a 100-mm capillary col-
umn (SP-2560; Supelco, Bellefonte, PA). Fatty acids of interest were
identified by comparison with known standards and expressed as a
percentage of total fatty acids. The coefficient of variation for the
RBC EPA DHA assays was 5%.
Data Analysis
All data are reported as means SE. The data were digitized (1
kHz) and recorded using a Biopac MP-100 data acquisition system
(Biopac Systems, Goleta, CA). The HR and HRV data were averaged
over 30-s intervals either during exercise or the coronary occlusion.
The following time points were evaluated for the exercise trials: the
last 30 s before the onset of exercise, from 0 to 30 s, from 30 to 60 s,
and from 90 to 120 s after exercise onset; and for recovery from
exercise: the last 30 s of exercise, from 0 to 30 s, from 30 to 60 s, and
from 90 to 120 s after the termination of exercise. The values averaged
over these time periods are reported as time 0, 30, 60, and 120 s,
respectively. ECG variables were averaged over the last five beats
before and 60 s after the onset of the coronary occlusion. QT interval
was corrected for changes in HR using Van de Waters correction
formula [QTc QT-87(60/HR-1)] (49).
The data were compared using ANOVA for repeated measures
(NCSS Statistical Software, Kaysville, UT). For example, the effects
the n-3 PUFAs on resting HR and HRV were analyzed using a
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H2290 n-3 FATTY ACIDS INCREASE HRV AT REST BUT NOT DURING METABOLIC STRESS

Table 1. Red blood cell omega-3 polyunsaturated fatty acid content

EPA DHA Omega-3 Index

Pre Post Pre Post Pre Post

Placebo (n 12) 0.16 0.01 0.25 0.04 0.28 0.07 0.32 0.06 0.43 0.08 0.54 0.08
1 g/day (n 6) 0.19 0.02 1.20 0.13* 0.17 0.02 2.15 0.22* 0.36 0.03 3.34 0.26*
2 g/day (n 12) 0.16 0.01 1.71 0.11* 0.29 0.08 2.01 0.13* 0.56 0.13 3.72 0.22*
4 g/day (n 22) 0.20 0.02 3.92 0.21* 0.21 0.03 3.02 0.17* 0.41 0.05 7.11 0.27*
Sham (n 5) 0.24 0.03 4.41 0.48* 0.17 0.02 3.421 0.33* 0.40 0.05 7.49 0.45*
All values are means SE and are expressed as %total lipid content. Sham (i.e., no myocardial infarction) received 4 g/day; EPA eicosapentaenoic acid;
DHA docasahexaenoic acid; omega-3 index EPA DHA. *P 0.01, pre vs. post.

two-way ANOVA [pre-post (2 levels) dose (4 levels)] with re- PUFA doses elicited similar reductions in HR, and increases
peated measures on one factor (pre-post) measures. In a similar HRV such that there were no significant differences noted

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fashion, the effects of n-3 PUFAs on the HR and HRV response to between these treatments (i.e., no significant dose effect: HR,
either submaximal exercise or the coronary artery occlusion at rest P 0.51; HRV: HF, P 0.66, SD, P 0.80 or dose
were analyzed using a three-factor ANOVA [pre-post (2 levels)
dose (placebo vs. n-3 PUFA) time (exercise 7 levels or occlusion
pre-post interaction: HR, P 0.28; HRV: HF, P 0.42, SD,
6 levels) with repeated measures on two factors (pre-post and time)]. P 0.10 ).
The effect of n-3 PUFA on ECG variables before and 60 s after Sham (no infarction) dogs: n-3 PUFA treatment (4 g/day)
coronary occlusion were evaluated using a three factor [pre-post (2 elicited reductions in HR (pre 124.8 6.3 vs. post 103.2
levels), dose (2 levels), and occlusion time (2 levels)] ANOVA with 10.8 beats/min) and increases in HRV (HF: pre 5.2 0.3 vs.
repeated measures on two (pre-post and occlusion time) factors. post 7.2 0.7 ln ms2; SD: pre 40.2 7.2 vs. post 78.4 20.8
Homogeneity of covariance (sphericity assumption, equal correlates ms) that were similar to those induced in the infarcted dogs.
between the treatments) was tested using Mauchleys test and, if Thus n-3 PUFA provoked changes in resting HR and HRV that
appropriate, adjusted using Huynh-Feldt correction. RBC and cardiac were consistent with an enhanced cardiac vagal regulation (2,
tissue lipid compositions were compared using a two-factor ANOVA 48) in both infarcted and noninfarcted animals.
with repeated measures on one factor (pre-post) or a one-factor
ANOVA, respectively. If the F value exceeded a critical value (P
Effect of n-3 PUFA on the ECG Variables, HR, and HRV
0.05), post hoc comparisons of the data were then made using
Tukey-Kramer multiple comparison test. Response to Acute Myocardial Ischemia
As with resting values, each dose of n-3 PUFA elicited
RESULTS
similar changes in the HR and HRV responses to the 2-min
Effect of n-3 PUFA on RBC and Cardiac Tissue Fatty Acid coronary artery occlusion (no significant dose effect: HR, P
Content 0.63; HRV: HF, P 0.54, SD, P 0.10). Therefore, the n-3
PUFA data for all three doses have been combined in the
In agreement with our previous study (11), n-3 PUFA subsequent analyses. The effects of placebo and the n-3 PUFA
supplements elicited a dose-dependent (dose effect, pre-post, treatment on the HR and HRV (HF only) response to the acute
and dose pre-post interaction all; P 106) increases in myocardial ischemia are displayed in Fig. 3, while the effects
RBC membrane EPA, DHA, and the omega-3 index (EPA of the coronary occlusion on ECG variables are listed in
DHA) compared with the placebo-treated animals (Table 1). Table 3. The coronary occlusion provoked significant increases
Thus n-3 PUFA treatment (all 3 doses) increased RBC n-3 in HR (P 106) and the descending portion of the T wave
PUFA content while lipid composition did not change during
the 3-mo study in the placebo-treated dogs. In a similar
manner, cardiac tissue n-3 PUFA content was significantly Table 2. Cardiac tissue omega-3 polyunsaturated fatty acid
higher (P 106, all 3 doses) in n-3 PUFA compared with the content
placebo-treated animals (Table 2).
Omega-3
EPA DHA Index
Effect of n-3 PUFA on Resting HR and HRV
Right atrium
As n-3 PUFA have been reported to elicit different responses placebo (n 12) 0.32 0.11 0.56 0.14 0.85 0.22
in males than in females (17), the effect of gender was first 1 g/day (n 6) 1.24 0.11* 3.38 0.20* 4.63 0.18*
2 g/day (n 12) 1.45 0.23* 2.89 0.41* 4.34 0.64*
evaluated. There were no significant gender differences in HR 4 g/day (n 22) 2.76 0.44* 3.54 0.50* 6.31 0.88*
(P 0.69) or HRV (HF, P 0.91; SD, P 0.30) before or Sham (n 5) 2.80 0.43* 3.83 0.23* 6.63 0.31*
after the 3-mo treatment period. Therefore, these data have Left ventricle
been combined in all subsequent analyses. The effects of placebo (n 12) 0.31 0.06 0.49 0.08 0.80 0.11
placebo and the n-3 PUFA treatment on resting (i.e., obtained 1 g/day (n 6) 1.25 0.09* 3.08 0.17* 4.33 0.21*
2 g/day (n 12) 1.69 0.20* 2.67 0.21* 4.35 0.64*
while the animals were standing waiting for the onset of 4 g/day (n 22) 3.81 0.30* 3.43 0.20* 7.24 0.45*
exercise) HR and HRV are displayed in Fig. 2. HR decreased Sham (n 5) 3.71 0.43* 3.65 0.28* 7.07 0.70*
(pre-post effect, P 0.006) and HRV [either HF, P 0.008,
All values are means SE and are expressed as %total lipid content. Sham
or SD, P 0.0006] increased after 3 mo of treatment while (i.e., no myocardial infarction) received 4 g/day; EPA eicosapentaenoic
these variables did not change during the 3-mo duration of the acid; DHA docasahexaenoic acid; omega-3 index EPA DHA. *P
study in the placebo (P 0.98) treated group. All three n-3 0.01, omega-3 treated vs. placebo.

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 n-3 FATTY ACIDS INCREASE HRV AT REST BUT NOT DURING METABOLIC STRESS
Fig. 2. Effect of increasing doses of dietary omega-3 polyunsaturated fatty
acids (n-3 PUFAs) on baseline heart rate and heart rate variability. Data were
obtained at rest and the last 30 s before the onset of a submaximal exercise test.
Vagal tone index is the high frequency component of the R-R interval
variability (0.24 to 1.04 Hz). SD, standard deviation of the R-R interval; pre,
before placebo or n-3 PUFA treatment began; post, after 3 mo of treatment.
Placebo (n 19), 1 g/day n-3 PUFA (n 6), 2 g/day n-3 PUFA (n 12), and
4 g/day n-3 PUFA (n 22) are shown. Each 1-g capsule of n-3 PUFA
contained 465 mg ethyl eicosapentaenoate (EPA) ethyl docosahexaenoate
(DHA) 375 mg while the placebo contained corn oil (1 g, 58% linoleic acid
28% oleic acid). *P 0.01, posttreatment vs. the corresponding pretreatment
values.
(P 105), a marker of the dispersion of repolarization (41,
51) while eliciting decreases in PR interval (P 106). No
other ECG variable was affected by the ischemia. The n-3
PUFA treatment (but not placebo) elicited significant reduc-
tions in HR (P 0.004) and increases in the PR interval (P
0.0005), changes that were maintained during the coronary
occlusion. Interestingly, preocclusion QTc interval increased in
both the placebo and n-3 PUFA treatment (P 105) at the
end of the 3-mo treatment period compared with values ob-
tained before the treatment began.
In agreement with previous studies (3, 18), the coronary
occlusion significantly increased HR (occlusion time effect,
P 106) and decreased both indexes of HRV (occlusion time
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H2291
effect, P 106) in the placebo and in the n-3 PUFA-treated
animals. HR was significantly lower (pre-post, P 0.0005),
and both HF (pre-post, P 0.00004) and SD (data not shown
pre-post, P 0.0008) were higher during the coronary occlu-
sion after n-3 PUFA treatment compared with values obtained
before the treatment. However, the absolute change in these
variables induced by myocardial ischemia (HR pretreatment
31.8 4.1 vs. posttreatment 28.1 4.5 beats/min; HF pre
3.6 0.4 vs. post 2.9 0.4 ln ms2; SD pre 43.8 5.1
vs. post 30.3 4.8 ms) was not altered by the n-3 PUFA
treatment (pre-post time interaction: HR, P 0.70; HF, P
0.11; SD, P 0.15). Thus n-3 PUFA treatment elicited a
downward shift in resting HR and an upward shift in the resting
HRV but did not alter the magnitude of the change in these
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variables that was induced by the coronary artery occlusion
(i.e., the response to the coronary occlusion per se was not
affected by the treatment). Similar results were obtained for the
sham n-3 PUFA-treated dogs (data not shown).
In contrast to the results from the n-3 PUFA treatment,
neither HR (pre-post, P 0.83), nor the HF component of R-R
interval variability (pre-post, P 0.20), nor the SD of R-R
interval (pre-post, P 0.14) was different before and at the
end the placebo treatment period. The change in these variables
was also similar before and at the end of the placebo treatment
(pre-post time interaction: HR, P 0.98; HF, P 0.57; SD,
P 0.67).
Effect of n-3 PUFA on the HR and HRV Response to
Submaximal Exercise
As with resting values, each dose of n-3 PUFA elicited
similar changes in the HR and HRV responses to exercise
(no significant dose effect: HR, P 0.88; HRV: HF, P
0.75, SD, P 0.83). Therefore, the n-3 PUFA data for all
three doses have been combined in all subsequent analyses.
As one would predict, exercise provoked large increases in
HR (exercise level effect, P 106) that were accompanied
by large reductions in HRV (exercise level effect, P
106). As these results confirm previous studies (3), these
data are not presented. Rather, the major focus in the sub-
sequent sections has been placed on the analysis of the effects
of n-3 PUFA treatment on the HR and HRV response to
exercise.
Submaximal exercise. The HR and HRV response to sub-
maximal exercise before and after 3 mo of placebo or n-3
PUFA treatment are displayed in Fig. 4. HR was significantly
lower (HR, pre-post effect P 0.03), and both HF (pre-post
P 0.02) and SD (data not shown pre-post P 0.00004) were
higher during exercise after n-3 PUFA treatment compared
with values obtained before the treatment. In contrast, neither
HR (pre-post HR, P 0.41) nor the SD of R-R interval
(pre-post SD, P 0.62) was altered while HF increased at the
end the placebo treatment period (pre-post, P 0.02). The HF
increase noted in the placebo-treated animals was similar to
that noted in the n-3 PUFA-treated dogs (i.e., HF were not
significantly different in placebo and n-3 PUFA-treated ani-
mals, P 0.66) while HR was significantly lower in the n-3
PUFA-treated as compared in the placebo-treated dogs (P
0.03). The changes in HR (pre 64.1 4.1 vs. post 63.3 45
beats/min) and HF (pre 5.1 0.2 vs. post 5.0 0.2 ln ms2)
provoked by exercise were not affected by the n-3 PUFA
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Fig. 3. Effect of n-3 PUFAs on the heart rate and heart rate variability response to acute myocardial ischemia (2-min left circumflex coronary artery
occlusion). As there were no significant differences noted between the various n-3 PUFA doses, these data have been combined. Dietary n-3 PUFA, but
not the placebo, produced a significant downward shift of the heart rate response that was accompanied by an upward shift in the high frequency
component of the R-R interval variability (vagal tone index: 0.24 1.04 Hz; i.e., significant pre-post treatment effect). Despite the shifts in the curves, the
absolute change in these variables induced by the occlusion was not altered before or after the treatment (i.e., there was no significant pre-post treatment
ischemia interaction). Placebo (n 15) and n-3 PUFA (1 4 g/day, n 39) are shown. Pre, before treatment began; post, after 3 mo of treatment; C,
control preocclusion; R, recovery postocclusion.

treatment (no significant pre-post exercise level interaction:
HR, P 0.72; HF, P 0.97). In contrast, exercise elicited a
significantly (pre-post exercise level interaction P 0.02)
greater reduction in SD; following n-3 PUFA treatment resting
SD increased (pre 54.4 3.6 vs. post 72.9 5.4 ms) but
declined to a similar value at the final stage of exercise both
before and after the n-3 PUFA treatment (pre 13.2 1.1 vs.
post 15.6 1.3 ms). Thus n-3 PUFA produced a shift in the
preexercise values of HR, HF, and SD that was maintained
throughout the exercise period. This treatment either did not
alter (HR and HF) or actually increased (greater change in SD,
pre 41.2 3.5 vs. post 57.3 5.0 ms) the magnitude of
the change in these variables that was induced by exercise. In
other words, the response to exercise per se was not affected by
the treatment. These data suggest that the HR reductions
induced by n-3 PUFA did not result solely from changes in
cardiac parasympathetic regulation. Similar results were ob-
tained for the sham n-3 PUFA-treated dogs. HR was decreased
and both HF and SD were increased at rest and during exercise
by n-3 PUFA treatment (data not shown).
Exercise onset. The changes (from preexercise levels) in HR
and HRV (HF component) provoked by onset of exercise
before and after 3 mo treatment with either the placebo or n-3
PUFA are displayed in Fig. 5. HR was significantly lower (HR,
pre-post effect P 0.04), and both HF (pre-post, P 0.02)
and SD (data not shown pre-post, P 0.0002) were higher
during the onset of exercise after n-3 PUFA treatment com-
pared with values obtained before the initiation of the treat-
ment. In contrast, neither HR (pre-post, P 0.27), HF (pre-
post, P 0.77), nor the SD of R-R interval (pre-post, P
0.74) was different before and at the end of the placebo
treatment period. In a similar manner, the change in HR, HF,
and SD provoked by exercise were not affected by either the
n-3 PUFA treatment (no significant pre-post exercise level
interaction: HR, P 0.39; HF, P 0.16; SD, P 0.10) or the
placebo treatment (no significant pre-post exercise: HR, P
0.28; HF, P 0.70; and SD, P 0.78). Thus, although n-3
PUFA produced a shift in the baseline (preexercise onset)
values of HR, HF, and SD, this treatment did not alter the
change in these variables induced by the onset of exercise.

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 n-3 FATTY ACIDS INCREASE HRV AT REST BUT NOT DURING METABOLIC STRESS H2293
Table 3. Effect of dietary omega-3 fatty acid on ECG reached before the treatment began). However, the absolute
parameters at rest and during coronary artery occlusion magnitude of the change in HR and HRV provoked by either
exercise or acute myocardial ischemia was not altered by
Pretreatment Posttreatment
n-3 PUFA treatment and was similar to that recorded for the
Control Occlusion Control Occlusion placebo. In other words, the n-3 PUFA treatment produced
Heart rate, beats/ parallel shifts in the response to either exercise or the
min coronary occlusion due to changes in resting (prechallenge)
Placebo 114.8 4.9 154.4 9.7* 115.0 6.0 152.8 11.1* HR and HRV. These data suggest that reductions in intrinsic
n-3 PUFA 122.0 3.5 153.8 5.0* 109.3 3.2 137.3 5.1* pacemaker rate rather than enhanced cardiac parasympa-
PR interval, ms
Placebo 95.1 3.2 83.6 4.1* 100.4 3.4 91.1 3.3* thetic activity may be responsible for the n-3 PUFA-induced
n-3 PUFA 97.2 2.4 87.8 2.5* 104.2 2.3 96.1 2.7* changes in HR. Fourth, a more rapid HR recovery following
QRS duration, ms the termination of exercise was noted after n-3 PUFA
Placebo 80.1 2.7 79.6 2.8 83.7 2.4 82.4 1.7 treatment compared with placebo-treated animals. However,
n-3 PUFA 81.4 1.3 79.2 1.5 82.2 0.9 81.5 1.1
QTc interval, ms
this faster HR recovery was not accompanied by corre-

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Placebo 248.1 4.3 251.3 3.8 270.4 3.7 258.9 3.7
n-3 PUFA 254.6 2.4 253.8 2.6 271.3 2.6 265.5 3.0
Tpeak-Tend
(corrected), ms
Placebo 88.0 3.8 95.4 4.0 88.8 2.9 101.4 5.5*
n-3 PUFA 93.1 2.3 105.4 2.6* 89.2 2.6 105.6 3.2*
Values are means SE. Heart rate correction: QTc, QT-87(60/HR-1);
Tpeak-Tend(corrected), Tpeak-Tend-87(60/HR-1) (Ref. 49). *P 0.01, con-
trol vs. coronary artery occlusion (60 s after occlusion onset). P 0.01,
pretreatment vs. posttreatment.
Similar results were obtained for the sham n-3 PUFA-treated
dogs (data not shown).
Exercise recovery. The changes in HR and HRV (HF)
recovery following the termination of exercise before and after
3-mo treatment with either the placebo or n-3 PUFA are
displayed in Fig. 6. Placebo treatment did not affect the
recovery of HR (pre-post, P 0.89), HF (P 0.11), or SD
(P 0.74) following the termination of exercise. In contrast,
HR declined more rapidly (i.e., a faster recovery, HR pre-post
P 0.001) after n-3 PUFA treatment compared with values
obtained before the initiation of the treatment. The faster HR
recovery from exercise after n-3 PUFA treatment was accom-
panied by a greater increase in SD (pre-post, P 0.001) but
not HF (pre-post, P 0.10). In a similar manner, absolute
change in HR (pre-post time interaction P 0.05) and in SD
(pre-post time interaction, P 0.03) following the termi-
nation of exercise were greater, while HF (no significant
pre-post time interaction, P 0.99) was not altered by n-3
PUFA treatment compared with values obtained before n-3
PUFA treatment. Similar results were obtained for the sham
n-3 PUFA-treated dogs (data not shown).
DISCUSSION
The present study investigated the effects of dietary n-3
PUFA (1 4 g/day for 3 mo) on HR and HRV both at rest
(baseline conditions) and during physiological stress (exer-
cise or acute myocardial ischemia). The major findings of
the study are as follows: first, and in agreement with
previous studies (11, 26), the n-3 PUFA treatment elicited
dose-dependent increases in both RBC and cardiac (right
atrial and left ventricular) tissue DHA and EPA content.
Second, n-3 PUFA treatment induced reductions in resting
HR that were accompanied by increases in HRV. Third,
these changes were maintained during a physiological chal-
lenge (the peak values obtained during the stimulus were
lower after n-3 PUFA treatment compared with values
sponding increases in the HF component of R-R interval
variability, data that would also suggest that cardiac vagal
regulation was not affected by the n-3 PUFA treatment.
Finally, similar HR and HRV changes were noted in sham
(noninfarcted animals), and as such, the n-3 PUFA changes
are not restricted to individuals with preexisting cardiac
damage.
Effect of n-3 PUFA on Resting HR and HRV
In agreement with the present study, a number of clinical
(1517) and experimental studies (11, 31, 33, 34) report that
n-3 PUFA ingestion or acute intravenous administration (9)
lower HR and increase HRV, suggestive of an increase in
cardiac parasympathetic regulation. The PR interval also in-
creased following n-3 PUFA treatment in the present study,
data also consistent with a parasympathetically mediated re-
duction in atrioventricular nodal conduction. However, there
are also studies in which n-3 PUFA failed to alter either HRV
or other measures of autonomic function (22, 24, 45), such as
baroreceptor sensitivity (22). Furthermore, even in the studies
that reported a positive action of n-3 PUFAs on HR or HRV,
the effect was often quite small (36 38). For example, a
meta-analysis of 30 trials found that fish oil supplements (3.5
g/day of EPA DHA) reduced baseline HR by 2.5 beats/min
(38), while Mozaffarian et al. (37) reported that individuals
with the highest fish consumption (5 meals per weak) only
exhibited 1.5-ms greater HRV compared with those with the
lowest fish consumption. Although this difference was statis-
tically significant, such a small change in resting HRV is not
likely to be physiologically relevant. Indeed, these investiga-
tors calculated that only a 1.1% reduction in the relative risk
for sudden cardiac death could be associated with this very
modest increase in HRV (37). However, these small changes
could have important consequences if they are maintained
during a physiological stressor such as exercise or acute myo-
cardial ischemia. Reductions in HR would reduce metabolic
demand placed on the heart particularly when oxygen supply is
compromised by coronary artery lesions/obstructions. The re-
sulting better match between oxygen supply and oxygen de-
mand would, indirectly, decrease the risk for adverse cardiac
events associated with myocardial ischemia. In fact, individu-
als with the lowest resting HRs also exhibited the lowest
long-term (20 yr) mortality rate (30). Furthermore, the ben-
eficial effects of -adrenergic receptor antagonists, the most
effective anti-arrhythmic medication, have been attributed to
the negative chronotropic actions of these drugs (27).

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Fig. 4. Effect of n-3 PUFAs on the heart rate and heart rate variability response to submaximal exercise. As there were no significant differences noted between
the various n-3 PUFA doses, these data have been combined. Dietary n-3 PUFA, but not the placebo, produced a significant downward shift of the heart rate
response curve that was accompanied by an upward shift in the high frequency component of the R-R interval variability (vagal tone index: 0.24 to 1.04 Hz;
i.e., significant pre-post effect). Despite the shifts in the curves, the absolute change in these variables induced by the exercise was not altered before or after
the treatment (i.e., there was no significant pre-post treatment exercise interaction). Data were averaged over the last 30 s of given exercise level. Exercise
levels were as follows: 1 0 kph and 0% grade; 2 4.8 kph and 0% grade; 3 6.4 kph and 0% grade; 4 6.4 kph and 4% grade; 5 6.4 kph and 8% grade;
6 6.4 kph and 12% grade; 7 6.4 kph and 16% grade. Pre, before placebo (n 16) or n-3 PUFA (1 4 g/day, n 35) treatment began; post, after 3 mo
of treatment.

Effect of n-3 PUFA on HR and Heart Variability Response
to Myocardial Ischemia
As far as we are aware, the present study provides the first
description of the effects of dietary n-3 PUFAs on the HR
and HRV response to acute myocardial ischemia in intact
unanesthetized preparations. As expected from previous
studies (3, 18), the coronary artery occlusion elicited a
robust HR increase that was accompanied by a rapid with-
drawal in parasympathetic regulation as indicated by the
decline in both total R-R interval variability (SD) and the
HF component of R-R interval variability. However, despite
alterations in preocclusion HR and HRV, the cardiac re-
sponse to coronary artery occlusion was not altered by the
n-3 PUFA treatment. The n-3 PUFA treatment produced
parallel shifts in the coronary occlusion response curves
(due to changes in the preocclusion HR and HRV), but the
magnitude of the change in these variables was not altered
by the n-3 PUFA treatment. As the robust autonomic re-
sponse to the coronary occlusion was not altered, the
changes in preischemic HR and HRV induced by the n-3
PUFA may be insufficient to prevent malignant changes in
the cardiac rhythm. Indeed, the results from the present
study are analogous to those obtained following treatment
with low doses of the cholinergic antagonist atropine (23).
With the use of the same canine model of myocardial
infarction, low-dose atropine decreased baseline HR and
increased HRV but, as in the present study, did not alter the
response to myocardial ischemia (23). This intervention also
failed to prevent the induction of ventricular fibrillation
(23). In marked contrast, however, exercise training not only

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Fig. 5. Effect of n-3 PUFAs on the change in heart rate and heart rate variability in response to the onset of submaximal exercise. As there were no significant
differences noted between the various n-3 PUFA doses, these data have been combined. As indicated in Fig. 4, dietary n-3 PUFA, but not the placebo, produced
a significant downward shift of the heart rate response that was accompanied by an upward shift in the high frequency component of the R-R interval variability
(vagal tone index: 0.24 1.04 Hz; i.e., significant pre-post treatment effect). Despite the shifts in the curves, the absolute change in these variables induced by
the exercise was not altered before or after the treatment (i.e., there was no significant pre-post treatment exercise onset interaction). Placebo (n 16) and
n-3 PUFA (1 4 g/day, n 35) are shown. Pre, before treatment began; post, after 3 mo of treatment.

improved resting HR and HRV but also dramatically re-
duced the response to coronary occlusion and completely
suppressed the formation of malignant ventricular tachyar-
rhythmias (7). When considered together, these results
strongly suggest that, to be effective, an intervention must
enhance cardiac parasympathetic regulation during myocar-
dial ischemia; resting changes alone may be insufficient to
protect against malignant arrhythmias.
Effect of n-3 PUFA on HR and Heart Variability Response
to Exercise
In the present study, dietary n-3 PUFA treatment also
elicited similar parallel shifts in the HR and HRV in the
exercise HR and HF response curves as was noted during the
coronary occlusion. However, exercise elicited a greater reduc-
tion in the SD in R-R interval following n-3 PUFA treatment,
and preexercise SD increased but declined to a similar value at
the final stage of exercise both before and after the n-3 PUFA
treatment. Interestingly, the HF component of R-R interval
variability exhibited a small but significant increase at the end
of the 3-mo placebo period, an increase similar to that noted
after n-3 PUFA treatment. Despite these similar changes in HF,
the HR response to exercise was reduced in n-3 PUFA com-
pared with placebo-treated dogs. These data suggest that the
HR reductions induced by n-3 PUFA did not result solely from
changes in cardiac parasympathetic regulation. In contrast to
these n-3 PUFA findings, an endurance exercise training pro-
gram (10-wk treadmill running) both improved resting HR and
HRV and attenuated the cardiac response to submaximal ex-
ercise in the same canine model as was used in the present
study (7, 8) .
There are relatively few studies that evaluated the effects of
n-3 PUFA on the response to exercise in humans (13, 39, 40,
43). In agreement with the present study, dietary fish oil

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Fig. 6. Effect of n-3 PUFAs on the change in heart rate and heart rate variability following the termination of submaximal exercise. As there were no significant
differences noted between the various n-3 PUFA doses, these data have been combined. Heart rate more rapidly declined following the termination of exercise
after the n-3 PUFA but not in the placebo-treated dogs. In contrast to what was noted during exercise onset, the absolute changes in heart rate following the
termination of exercise were greater after dietary n-3 PUFA treatment (i.e., significant pre-post treatment exercise recovery interaction). However, the more
rapid decline in heart rate was not accompanied by an upward shift in the high frequency component of the R-R interval variability (vagal tone index: 0.24 1.04
Hz; i.e., no significant pre-post treatment exercise recovery interaction). Placebo (n 16) and n-3 PUFA (1 4 g/day, n 35) are shown. Pre, before treatment
began; post, after 3 mo of treatment. *Posttreatment vs. the corresponding pretreatment time interval.

supplements (8-wk, 8 g/day) produced HR reductions both at
rest and at peak workload in well-trained men (bicyclists)
during submaximal exercise (43). In a similar manner, fish oil
(5-wks, 6 g/day) reduced HR during exercise in a group of
Australian rule football players (13). Finally, fish oil supple-
ments (12 wk, 6 g/day DHA-rich tuna oil) produced small
reductions in baseline HR and larger reductions in HR with
increasing workload in sedentary overweight adults (39). Un-
like the previous clinical studies (13, 40, 43), these investiga-
tors also evaluated the effects of the interventions on HRV
(39). The fish oil supplement also increased the HF component
of HRV both at rest and during exercise. The changes in HR
and HRV induced by the fish oil supplements were similar to
those recorded in a group of exercise-trained subjects (39). The
authors concluded that fish oil supplements reduced HR and
modulated HRV in keeping with an improved parasympathetic-
sympathetic balance in overweight adults (39). These latter
results contrast somewhat with the present study, in that the
tuna oil supplements induced modest changes in baseline HRV
and HR but also attenuated both the HR and the HRV re-
sponses to exercise. Species (dog vs. human) may account for
the seemingly disparate results.
Analysis of the HR and HRV response to the onset and
recovery from exercise can also provide insight into cardiac
autonomic regulation (4, 47). Although preexercise onset HR
decreased and HRV increased at the end of the 3-mo n-3 PUFA
treatment, the changes in these variables that were induced by
exercise onset were similar both before and after treatment. As
far as we are aware, no other studies (either in humans or
animals) have evaluated the effects of n-3 PUFA on the cardiac
response to the onset of exercise. Once again, the findings
reported in the present study contrast with the results obtained
following a 10-wk exercise training program (8). Exercise
training significantly attenuated the increase in HR but did not
alter the reductions in HRV, elicited by the onset of exercise,
data that suggest that cardiac sympathetic rather than cardiac
parasympathetic response to exercise onset was reduced fol-
lowing exercise training (8).
In contrast to the response to exercise onset, n-3 PUFA
treatment altered both the HR and SD of R-R interval recovery

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 n-3 FATTY ACIDS INCREASE HRV AT REST BUT NOT DURING METABOLIC STRESS
following termination of exercise. In agreement with human
studies (40), HR declined more rapidly (i.e., a faster recovery),
which was accompanied by a greater increase in total R-R
interval variability but not the HF component of this variability
(an index of cardiac parasympathetic activity) after n-3 PUFA
treatment compared with values obtained before the initiation
of the treatment. Thus the absolute changes in HR and the SD
of R-R interval following the termination of exercise were
greater, while the HF component of HRV was not altered by
n-3 PUFA treatment compared with values obtained before n-3
PUFA treatment. Since the HF component of R-R interval
variability was not altered by n-3 PUFA treatment, the faster
return to baseline after termination of exercise in the n-3
PUFA-treated animals probably did not result as a consequence
of a faster reactivation of cardiac parasympathetic activity but
rather may reflect either changes in the intrinsic pacemaker rate
or cardiac sympathetic activity.
Possible Mechanism of Action
The mechanisms responsible for the n-3 PUFA-induced
decreases in HR and increases in HRV were not investigated in
the present study and remain to be determined. However, there
are at least two possible explanations for these observations:
HR reductions could result from changes in cardiac autonomic
regulation (enhanced parasympathetic and/or reduced sympa-
thetic activity) and/or from alterations in intrinsic pacemaker
rate. In the present study, the prechallenge (exercise or coro-
nary occlusion) HF component of the R-R interval variability
increased following 3 mo of n-3 PUFA treatment. Although
there are limitations with this index (2, 28, 42), it is now
generally accepted that it can provide an indirect and qualita-
tive assessment of cardiac parasympathetic regulation (2, 48).
As such, the reduction in HR that accompanies n-3 PUFA
treatment could reflect an enhanced parasympathetic efferent
(either of central or peripheral origin) regulation of the heart.
However, as has been noted, both exercise and coronary artery
occlusion elicited similar changes in HRV before and after n-3
PUFA treatment. The parallel shifts in the HRV response
curves after n-3 PUFA treatment are more consistent with
reductions in the intrinsic pacemaker rate than with alterations
in autonomic neural regulation. If parasympathetic activity had
been enhanced or sympathetic activity reduced following n-3
PUFA treatment, then one would expect smaller changes in HR
and HRV during physiological interventions (as was the case
following endurance exercise training; Refs. 7, 8) rather than
similar changes as noted in the present study. Furthermore, an
accelerated HR recovery was noted after n-3 PUFA without
corresponding changes in HF power. Finally, despite similar
changes in HRV, the peak HR response to exercise was
attenuated in the n-3 PUFA treated but not in the placebo-
treated dogs. When considered together, these data suggest that
intrinsic changes in pacemaker rate may play a more important
role in n-3 PUFA-induced HR reductions than do changes in
cardiac autonomic regulation. Indeed, n-3 PUFA have been
recently reported to reduce the pacemaker current (If) in sino-
atrial node cells obtained from rabbits fed diets enriched with
fish oil but not in cells isolated from the hearts of animals fed
sunflower oil (50).
AJP-Heart Circ Physiol VOL
H2297
Limitations of the Study
In the present study, cardiac vagal nerve activity was not
directly recorded. Cardiac parasympathetic regulation was only
indirectly evaluated using noninvasive markers of HRV. Al-
though a number of studies provide strong evidence that
beat-to-beat fluctuation in HR reflect corresponding changes in
cardiac parasympathetic regulation (2, 48), an accurate assess-
ment of nerve activity can only be obtained from direct nerve
recordings. As such, HRV data should always be interpreted
with care. Second, previous studies (10, 11) demonstrate that
ventricular function is not altered by myocardial infarction in
the canine model used in the present study. As such, one might
speculate that the potential benefits of dietary n-3 PUFA on
cardiac autonomic regulation could be more obvious in indi-
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viduals with more severe cardiac impairment. To the best of
our knowledge, there have not been any studies that have
evaluated the effects of dietary n-3 PUFA on the HRV re-
sponse to exercise or acute myocardial ischemia in patients
with impaired cardiac function. Future investigation will be
required to determine if n-3 PUFA exhibit greater effects in
these patient populations. Third, selecting an appropriate dose
can prove to be problematic when comparing across species of
different sizes. In the present study, if one adjusts dose by body
surface area, then the 1, 2, and 4 g/day dose would be
equivalent to 1.5, 4.9, and 9.4 g/day in human subjects. As
such, these doses are somewhat higher than the dose (1 g/day)
most commonly used in secondary prevention trials. However,
doses up to 8 g/day n-3 PUFA have been used to evaluate the
effect of n-3 PUFA on HRV in human subjects (43). In
addition, n-3 PUFA treatment in the present study yielded RBC
membrane concentrations within the range that were associated
with a significant reduction in the risk for sudden cardiac death
in epidemiological studies (1). Specifically, Albert et al. (1)
found that a mean RBC concentration of 6.9% was associated
with a 90% reduction in the risk for sudden death, a value that
compares favorably to that obtained in the present study (after
3 mo, the 4 g/day treatment RBC concentration was 7.1%).
Finally, although dogs and humans exhibit a similar cardiac
autonomic regulation (46), species difference could also con-
tribute to responses differences. One must always use caution
when extrapolating results between species.
Conclusions and Future Directions
In the present study, dietary n-3 PUFA (DHA EPA ethyl
esters, 1 4 g/day for 3 mo) elicited dose-independent reduc-
tions in baseline HR that were accompanied by increases in
HRV in dogs with and without healed myocardial infarctions.
However, n-3 PUFA treatment did not alter the robust auto-
nomic response (identical increases in HR and decreases in
HRV before and after treatment) induced by either exercise or,
more importantly, by myocardial ischemia. Furthermore, HR
recovery following exercise was faster after n-3 PUFA without
corresponding changes HF power and the peak HR obtained
during exercise was lower in n-3 PUFA-treated dogs compared
with placebo-treated animals despite similar HRV changes.
When considered together, these data suggest that changes in
cardiac autonomic regulation are not solely responsible for n-3
PUFA-induced changes in HR; alterations in the intrinsic
pacemaker rate also contribute to the chronotropic actions of
these fatty acids.
300 JUNE 2011 www.ajpheart.org
H2298 n-3 FATTY ACIDS INCREASE HRV AT REST BUT NOT DURING METABOLIC STRESS
If n-3 PUFA-induced changes in intrinsic pacemaker rate are
responsible for (or at least contribute to) the HR reductions
associated with this treatment, then one can make several
predictions. In vivo: 1) One would predict that selective an-
tagonists of the pacemaker current (If) should elicit parallel
shifts in the HR response to exercise or coronary occlusion
similar to those noted in the present study. In fact, Zatebradine
reduced both the resting HR and the HR at the end of each
stage of an exercise stress test (21). 2) One would predict that
after complete cardiac autonomic neural blockade, HR would
be lower in n-3 PUFA treated than in placebo-treated subjects.
If, however, intrinsic rate is not altered by this intervention,
then alterations in autonomic regulation could play an impor-
tant role in mediating the HR reduction following n-3 PUFA
treatment. In vitro: 3) One would predict a reduced If current
density in sinus nodal myocytes obtained from n-3 PUFA-
treated dogs similar to that which has been reported in rabbit
preparations (50). 4) This reduced pacemaker current could
result from either reduced expression of the pacemaker channel
(i.e., reduced HCN protein content) or from the posttransla-
tional modification of this protein (trafficking, phosphoryla-
tion, oxidation, etc). 5) In contrast, if the HR reduction resulted
from increased parasympathetic activity, then one would ex-
pect an increase in the acetylcholine-activated potassium cur-
rent (IK-ach) accompanied by an increased expression of this
channel protein and/or the muscarinic (M2) receptors (and
associated signaling pathway proteins). The determination of
the relative contribution of putative changes in intrinsic pace-
maker rate and cardiac autonomic regulation to n-3 PUFA-
mediated reductions in HR will require further investigation.
ACKNOWLEDGMENTS
We thank Raven Morgan, Anita McKenzie, and Andrew Christianson for
excellent technical assistance during the course of the studies. We also thank
GlaxoSmithKline for generously providing the n-3 PUFA ethyl ester and
placebo capsules for this study.
GRANTS
This work was supported by National Heart, Lung, and Blood Institute
Grant HL-086700 (to G. E. Billman).
DISCLOSURES
William S. Harris is a scientific advisor to GlaxoSmithKline, Monsanto, and
Unilever; a speaker for GlaxoSmithKline; and the owner of OmegaQuant, a
company offering blood omega-3 testing. There are no other conflicts to
declare.
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