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Protein carbonyl assay to measure oxidative stress in muscle of exercising horses supplemented with vitamin E

Published online by Cambridge University Press:  02 June 2009

K J Duberstein ,
S E Johnson ,
L R McDowell  and
E A Ott
K J Duberstein*
Affiliation:
Department of Animal Sciences, University of Florida, Gainesville, FL32611, USA
S E Johnson
Affiliation:
Department of Animal Sciences, University of Florida, Gainesville, FL32611, USA
L R McDowell
Affiliation:
Department of Animal Sciences, University of Florida, Gainesville, FL32611, USA
E A Ott
Affiliation:
Department of Animal Sciences, University of Florida, Gainesville, FL32611, USA
*
*Corresponding author: kjleejo@uga.edu
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Abstract

Intense exercise has been associated with free radical damage that forms potentially measurable by-products in the blood and muscle of exercising subjects. The extent of damage to the exercising animal has yet to be conclusively determined, and studies often focus on by-products in the blood rather than muscle. The current study examined the presence of oxidative products in the muscle of exercising horses as well as the effects of excess vitamin E on the presence of these products. Eight Thoroughbred horses were used in a crossover design, with one group being fed vitamin E at the 1989 NRC [National Research Council (1989) Nutrient Requirements of Horses. 5th revised edn.; Washington DC: National Academy Press, pp. 48] level recommended for horses in moderate to intense work (80 IU kg DM− 1), and the second group being fed the control diet plus 3000 IU day− 1dl-α-tocopheryl acetate. The horses underwent an 8-week training programme and a final standard exercise test (SET). During the SET, the horses ran on a 6° incline to exhaustion. Muscle samples were biopsied before and after performing the SET and analysed for the presence of carbonyl groups and ubiquitin. Blood was collected prior to the SET and analysed for vitamin E. No significant differences in plasma vitamin E were found between treatment groups. However, myofibril carbonylation, a product of free radical damage to muscle tissue, was found to be lower in vitamin E-supplemented horses post-SET exercise (P < 0.05), suggesting that vitamin E influences some measures of oxidative stress in exercising horses, particularly following a strenuous bout of exercise. Ubiquitin was not detected in myofibrils, indicating clearance of carbonyl groups by a different mechanism.

Keywords

vitamin Eoxidative stressexerciseequinecarbonylmuscle damage
Type
Short Communication
Information
Comparative Exercise Physiology , Volume 6 , Issue 1 , February 2009 , pp. 1 - 5
Copyright
Copyright © Cambridge University Press 2009

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Footnotes

†

Present address: Edgar Rhodes Center For ADS, University of Georgia, 425 River Road, Athens, GA 30 602, USA

References

1 Powers, SK and Hamilton, K (1999). Antioxidants and exercise. Clinical Sports Medicine 18: 525–536.CrossRefGoogle ScholarPubMed
2 Avellini, L, Chiaradia, E and Gaiti, A (1999). Effect of exercise training, selenium and vitamin E on some free radical scavengers in horses (Equus caballus). Comparative Biochemistry and Physiology – Part B: Biochemistry and Molecular Biology 123: 147–154.CrossRefGoogle ScholarPubMed
3 Barreiro, E, Gea, J, Di Falco, M, Kriazhev, L, James, S and Hussain, S (2005). Protein carbonyl formation in the diaphragm. American Journal of Respiratory Cell Molecular Biology 32: 9–17.CrossRefGoogle ScholarPubMed
4 Goto, S, Nakamura, A, Radak, Z, Nakamoto, H, Takahashi, R, Yasuda, K, Sakurai, Y and Ishii, N (1999). Carbonylated proteins in aging and exercise: immunoblot approaches. Mechanisms of Ageing and Development 107: 245–253.CrossRefGoogle ScholarPubMed
5 Radak, Z, Kaneko, T, Tahara, S, Nakamoto, H, Ohno, H, Sasvari, M, Nyakas, C and Goto, S (1999). The effect of exercise training on oxidative damage of lipids, proteins, and DNA in rat skeletal muscle: evidence for beneficial outcomes. Free Radical Biology and Medicine 27: 69–74.CrossRefGoogle ScholarPubMed
6 Radak, Z, Takahashi, R, Kumiyama, A, Nakamoto, H, Ohno, H, Ookawara, T and Goto, S (2002). Effect of aging and late onset dietary restriction on antioxidant enzymes and proteasome activities, and protein carbonylation of rat skeletal muscle and tendon. Experimental Gerontology 37: 1423–1430.CrossRefGoogle ScholarPubMed
7 National Research Council, (1989). Nutrient Requirements of Horses. 5th Revised edn.; Washington, DC: National Academy Press, pp. 48.Google Scholar
8 Gordillo, GM, Atalay, M, Roy, S and Sen, CK (2002). Hemangioma model for in vivo angiogenesis: Inducible oxidative stress and MCP-1 expression in EOMA cells. Methods in Enzymology 352: 422–432.CrossRefGoogle ScholarPubMed
9 Kinnunen, S, Hyyppa, S, Lappalainen, J, Oksala, N, Venojarvi, M, Nakao, C, Hanninen, O, Sen, CK and Atalay, M (2005). Exercise-induced oxidative stress and muscle stress protein responses in trotters. European Journal of Applied Physiology 93: 496–501.CrossRefGoogle ScholarPubMed
10 Kinnunen, S, Hyyppa, S, Lehmuskero, A, Oksala, N, Maenpaa, P, Hanninen, O and Atalay, M (2005). Oxygen radical absorbance capacity (ORAC) and exercise-induced oxidative stress in trotters. European Journal of Applied Physiology 95: 550–556.CrossRefGoogle ScholarPubMed