Nutrition and Disease
Antioxidant Status in Dogs with Idiopathic Dilated Cardiomyopathy1,2
Lisa M. Freeman,3 Don J. Brown and John E. Rush
Department of Medicine, Tufts University School of Veterinary Medicine, North Grafton, MA 01536
EXPANDED ABSTRACT
KEY WORDS: ● antioxidant ● dilated cardiomyopathy
Free radicals, metabolites of oxygen metabolism, are pro-
duced under normal conditions, but their rate of production
does not exceed the capacity of the body to catabolize them. It
is only when the natural defenses are overwhelmed that free
radical damage occurs. Therefore, the host’s endogenous anti-
oxidant system plays a major role in the prevention or limi-
tation of myocardial damage. Endogenous antioxidants in-
clude enzymatic antioxidants (e.g., superoxide dismutase or
glutathione peroxidase), free radical scavengers (e.g., vitamins
A, C or E) and metal chelators. Antioxidants also can be
derived exogenously through the diet or through the use of
supplements.
Free radical-induced injury has been implicated in the
development of a number of cardiac diseases, including coro-
nary artery disease, myocardial infarction and some forms of
cardiomyopathy in people and laboratory animals (Kaul et al.
1993). Free radicals not only have cytotoxic effects on the
myocardium, but also act as negative inotropes (Prasad et al.
1993). Altered antioxidant status has been identified in an
aortic banding model of congestive heart failure, with elevated
levels during cardiac hypertrophy and decreased levels in fail-
ure (Dhalla and Singal 1994, Gupta and Singal 1989). Similar
changes have been seen in other human diseases such as
inflammatory bowel disease and human immunodeficiency vi-
rus (Delmas-Beauvieux et al. 1996, Hoffenberg et al. 1997).
Elevated mean vitamin A concentration was found in cats
with dilated cardiomyopathy compared with healthy controls
(Fox et al. 1993). Whether these alterations contribute to
disease progression or reflect a compensatory response to in-
creased free radical stress is unclear at present.
The purpose of this pilot study was to determine antioxi-
dant status in dogs with idiopathic dilated cardiomyopathy
(IDCM)4 compared with healthy controls.
1
Presented as part of the Waltham International Symposium on Pet Nutrition
and Health in the 21st Century, Orlando, FL, May 26 –29, 1997. Guest editors for
the symposium publication were Ivan Burger, Waltham Centre for Pet Nutrition,
Leicestershire, UK and D’Ann Finley, University of California, Davis.
2
Supported in part by Hill’s Pet Nutrition.
3
To whom correspondence should be addressed.
4
Abbreviations used: ACEI, angiotensin converting-enzyme inhibitor; IDCM,
idiopathic dilated cardiomyopathy; NYHA, New York Heart Association.
0022-3166/98 $3.00 © 1998 American Society for Nutritional Sciences. J. Nutr. 128: 2768S–2770S, 1998.
2768S
● glutathione peroxidase ● dogs
Materials and methods. Antioxidant status in canine
IDCM. All dogs were client-owned animals. A diagnosis of
IDCM was based on the presence of left atrial enlargement and
a fractional shortening ,28% (,22% in Doberman pinschers)
on 2-D and M-mode echocardiography. Controls were age-
Downloaded from jn.nutrition.org by on October 23, 2006
and weight-matched to the IDCM dogs. Dogs with major
concurrent diseases, such as cancer, chronic renal failure and
hepatic failure were excluded from the study. Owners signed a
consent form before enrolling their dogs in the study. The
study was approved by the Tufts University Animal Care and
Use Committee.
Blood (10 mL) was collected in EDTA, centrifuged and
separated within 20 min. Fasting levels of vitamins A (reti-
nol), C (ascorbic acid) and E (b-tocopherol) were measured in
plasma; glutathione peroxidase and superoxide dismutase were
measured in washed erythrocytes. Vitamins A, C and E were
determined by reverse-phase HPLC, and glutathione peroxi-
dase was analyzed using a Cobas Fara II centrifugal analyzer
(Roche Diagnostics Systems, Nutley, NJ). Superoxide dis-
mutase was determined by a commercial spectrophotometric
assay (SOD-525, Bioxytech, Cedex, France). Dietary vitamins
A and E were calculated based on manufacturer’s data (IU/kg
diet on a dry matter basis) for dogs eating commercial dog
foods. One dog, which ate a homemade diet, was excluded
from determinations of dietary vitamins. Mean antioxidant
concentrations between the IDCM and control groups were
compared using Student’s t test; Pearson correlation was used
to identify potential correlations between disease severity and
antioxidant concentrations. Results were considered signifi-
cant when the two-tailed P- value was , 0.05.
Results. Twelve dogs with IDCM and 11 healthy controls
were enrolled in the study. Mean age of the IDCM dogs was
8.9 6 2.5 y compared with 7.9 6 1.9 y in the control group (P
5 0.21). Body weight also was not different between the
groups (46.0 6 18.3 kg for the IDCM group vs. 40.2 6 6.9 kg
for the controls; P 5 0.57). All control dogs and 11 of the 12
IDCM dogs ate commercial dry diets; one IDCM dog ate a
homemade diet. In the IDCM group, one dog was classified as
New York Heart Association (NYHA) Class I, four were
NYHA Class II, five were NYHA Class III, and two were
NYHA Class IV. An arrhythmia was detected in 11 of 12
IDCM dogs (and 0 of 11 controls). The arrhythmia was atrial
 ANTIOXIDANT STATUS IN CANINE IDCM 2769S

TABLE 1
Mean echocardiographic measurements in dogs with
idiopathic dilated cardiomyopathy (n 5 12).

Mean 6 SD

Left atrium, cm 3.5 6 0.7

Aorta, cm 2.9 6 0.4
Left ventricular internal dimension in diastole, cm 5.7 6 0.9
Left ventricular internal dimension in systole, cm 4.6 6 0.9
End-diastolic volume index,1 mL/m2 175 6 118
End-systolic volume index,2 mL/m2 86 6 54
Fractional shortening, % 18.1 6 6.3
1
(Left ventricular internal dimension in diastole)3/body surface area.
2
(Left ventricular internal dimension in systole)3/body surface area.

fibrillation in six dogs and ventricular premature complexes in

five dogs. Medication regimens included an angiotensin-con-
verting enzyme inhibitor (ACEI) and a b-blocker (n 5 3);
ACEI, furosemide, digoxin and a b-blocker (n 5 6); and
ACEI, furosemide, digoxin and diltiazem (n 5 3). Mean

Downloaded from jn.nutrition.org by on October 23, 2006

echocardiographic measurements in the IDCM group are
shown in Table 1. FIGURE 1 Comparison of disease severity [based on end-systolic
Circulating antioxidant levels are shown in Table 2. Eryth- volume index (ESVI)] and glutathione peroxidase (GPX) concentrations
rocyte glutathione perioxidase concentrations were measured in dogs with idiopathic dilated cardiomyopathy (n 5 11) and controls
in only 21 of 23 dogs due to technical difficulties (11 dogs with (n 5 10); *ESVI 5 (left ventricular internal dimension in systole)3/body
IDCM and 10 controls). Mean erythrocyte glutathione perox- surface area.
idase concentration was significantly higher in IDCM dogs
(159.5 6 19.8 U/g Hb) compared with controls (138.4 6 14.4
York Heart Association Class was associated with plasma
U/g Hb; P 5 0.01). There was a trend for higher concentra-
vitamin C (r 5 0.49; P 5 0.02). Class also was correlated
tions of plasma vitamin A and vitamin C in the IDCM group
with dietary vitamin A (r 5 0.47; P , 0.05). There was no
as well, but these did not reach significance (P 5 0.12 and P
significant correlation between disease severity and plasma
5 0.11, respectively). There was no difference between groups
vitamin A, plasma vitamin E, erythrocyte superoxide dis-
in plasma vitamin E or erythrocyte superoxide dismutase.
mutase or dietary vitamin A.
There was a trend for higher levels of dietary vitamin A in the
Discussion. The results of this pilot study suggest that
IDCM group (32488 6 11481 IU/kg) compared with the con-
alterations in some aspects of the endogenous antioxidant
trols (22775 6 3005 IU/kg; P 5 0.07), although there was no
system exist in dogs with IDCM, especially for circulating
correlation between dietary vitamin A and circulating vitamin
glutathione peroxidase and vitamin C. These results con-
A. There was no difference between groups in dietary vitamin
tradict those found using a rodent model of cardiac hyper-
E levels (343 6 222 IU/kg for IDCM dogs vs. 275 6 124 IU/kg
trophy, in which reduced levels of antioxidants were found
for controls; P 5 0.48).
in animals with congestive heart failure (Gupta and Singal
Disease severity (NYHA Class) was related to circulating
1989). The cause for this discrepancy is unknown, but it is
antioxidants. There was a significant correlation between
likely related to the disease model because at least two
disease severity and erythrocyte glutathione peroxidase,
human studies demonstrated elevated antioxidant concen-
both when severity was measured by NYHA Class (r 5
trations in inflammatory bowel disease and human immu-
0.52, P 5 0.02) and when severity was measured by end-
nodeficiency virus (Delmas- Beauvieux et al 1996, Hoffen-
systolic volume index (r 5 0.60; P 5 0.009; Fig. 1). New
berg et al 1997). Whether these alterations, similar to those
in our study, are a primary or secondary occurrence is
currently unknown. Elevations in antioxidant levels may be
TABLE 2 a marker of a compensatory response to increased oxidant
Mean circulating antioxidant concentrations in dogs with stress. The elevations also may be secondary to medications
used in the therapy of IDCM. On the other hand, these
idiopathic dilated cardiomyopathy (IDCM) and controls1
alterations in antioxidant status may play a role in the
IDCM Controls P- development or progression of disease. Either way, further
(n 5 12) (n 5 11) value study of these changes is warranted.
This study also identified trends for higher dietary intakes of
Vitamin A, mol/L 4.0 6 1.7 3.1 6 1.1 0.12 vitamin A in dogs with IDCM and a correlation between
Vitamin C, mol/L 43.15 6 16.47 34.07 6 7.38 0.11 dietary vitamin A and disease severity. The most likely cause
Vitamin E, mol/L 49.6 6 23.1 50.4 6 18.8 0.92 for these weak relationships is that as cardiac disease becomes
Superoxide dismutase, more severe, a reduced-sodium cardiac diet is more likely to be
U/g Hb 2280 6 470 2071 6 412 0.27
Glutathione peroxidase, prescribed. These diets tend to contain high levels of vitamin
U/g Hb 159.5 6 19.8 138.4 6 14.4 0.01 A. Additional studies will be necessary to determine whether
circulating levels of antioxidants accurately reflect tissue con-
1
Values are mean 6 SD. centrations. For example, there are known to be some differ-
2770S SUPPLEMENT
ences in the myocardial vs. the circulating antioxidant system
because catalase is produced only in very low levels in the
heart (Janssen et al. 1993).
In summary, dogs with IDCM had elevated concentrations
of erythrocyte glutathione peroxidase, which were correlated
with disease severity. Plasma vitamin C and dietary vitamin A
also correlated with disease severity. Future studies will be
required to determine the significance of these findings, which
may be secondary to IDCM and its therapy or may play an
important role in the development and progression of the
disease.
LITERATURE CITED
Delmas-Beauvieux, M, Peuchant, E., Couchouron, A., Constans, J., Sergeant, C.,
Simonoff, M., Pellegrin, J., Leng, B., Conri, C. & Clerc, M. (1996) The
enzymatic antioxidant system in blood and glutathione status in human
immunodeficiency virus (HIV)-infected patients: effects of supplementation
with selenium or b-carotene. Am. J. Clin. Nutr. 64: 101–107.
Dhalla, A. K. & Singal, P. K. (1994) Antioxidant changes in hypertrophied and
failing guinea pig hearts. Am. J. Physiol. 266: H1280 –H1285.
Fox, P. R., Trautwein, E. A., Hayes, K. C., Bond, B. R., Sisson, D. D. & Moise, N. S.
(1993) Comparison of taurine, a-tocopherol, retinol, selenium, and total
triglycerides and cholesterol concentrations in cats with cardiac disease and
in healthy cats. Am. J. Vet. Res. 54: 563–569.
Gupta, M. & Singal, P. K. (1989) Higher antioxidant capacity during a chronic
stable heart hypertrophy. Circ. Res. 64: 398 – 406.
Hoffenberg, E. J., Deutsch, J., Smith, S. & Sokol, R. J. (1997) Circulating
antioxidant concentrations in children with inflammatory bowel disease.
Am. J. Clin. Nutr. 65: 1482–1488.
Janssen, M., Van der Meer, P. & De Jong, J. W. (1993) Antioxidant defenses
in rat, pig, guinea pig, and human hearts: comparison with xanthine oxi-
doreductase activity. Cardiovasc. Res. 27: 2052–2057.
Kaul, N., Siveski-Iliskovic, N., Hill, M., Slezak, J. & Singal, P. K. (1993) Free
radicals and the heart. J. Pharmacol. Toxicol. Methods 30: 55– 67.
Prasad, K., Kalra, J. & Bharadwaj, L. (1993) Cardiac depressant effects of
oxygen free radicals. Angiology 44: 257–270.
Downloaded from jn.nutrition.org by on October 23, 2006