Proteomic analysis emphasizes the adaptation of energy metabolism in horses during endurance races.
- Journal Article
Summary
The research study aims to analyze the effects of strenuous long-term aerobic exercise, specifically endurance racing, on the energy metabolism adaptation in horses. This investigation involves notably fluctuating serum protein levels before and after a race and provides insights into its role in maintaining physiological balance.
Methodology and Procedure
In order to monitor the alterations in horse physiology during the endurance race, the researchers:
- Collected serum samples from 13 endurance horses before and after an 80-km race.
- Conducted a proteomic analysis of these samples using a TMT-based quantitative method.
- Validated the apolipoprotein and haptoglobin values in the serum through enzyme-linked immunosorbent assay and biochemical assay respectively.
- Tracked difference in protein abundance between pre- and post-race values.
Results
The research findings include:
- Identification of 10 core proteins with significant differences in their pre- and post-race abundance levels in the serum samples.
- Post-race increase in protein abundance for the apolipoprotein groups (ApoA IV and E), Microfibril-associated glycoprotein 4 (MFAP4), transferrin, and antithrombin-III.
- Post-race decrease in apolipoprotein C-II, C-III and R, alpha-1-microglobulin/bikunin precursor protein (AMBP), and haptoglobin abundance.
- The Gene Ontology analysis demonstrated changes in areas such as triglyceride and acylglycerol homeostasis, lipid localization regulation, triglyceride catabolic processes, cholesterol binding, and antioxidant activity.
Conclusion
The endurance race bred several homeostatic imbalances marked by notable alterations in the levels of serum proteins. The most remarkable changes highlight the kind of adaptation that a horse’s energy metabolism undergoes, shifting towards consuming lipids more proficiently. These findings provide a matter-of-fact understanding of horse physiology under stress and could potentially guide better endurance race preparation and recovery strategies.
Cite This Article
Publication
Researcher Affiliations
- Clinic for Internal Diseases, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia. jselanec@vef.hr.
- Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia.
- Department of Chemistry and Biochemistry, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia.
- Department of Chemistry and Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, Zagreb, Croatia.
- Mediterranean Institute for Life Science (MedILS), University of Split, Split, Croatia.
- Clinic for Internal Diseases, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia.
- Clinic for Internal Diseases, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia.
- Clinic for Surgery, Orthopaedics and Ophthalmology, Faculty of Veterinary Medicine, University of Zagreb, Zagreb, Croatia.
MeSH Terms
- Animals
- Horses / physiology
- Energy Metabolism / physiology
- Proteomics
- Physical Endurance / physiology
- Physical Conditioning, Animal / physiology
- Male
- Adaptation, Physiological
- Haptoglobins / metabolism
- Haptoglobins / analysis
- Female
- Proteome / metabolism
- Apolipoproteins / metabolism
- Apolipoproteins / blood
Grant Funding
- 4135 / Croatian Science Foundation
- 4135 / Croatian Science Foundation
- 4135 / Croatian Science Foundation
- 4135 / Croatian Science Foundation
- 4135 / Croatian Science Foundation
- 621394 / European Commission FP7
- 621394 / European Commission FP7
- 621394 / European Commission FP7
- 621394 / European Commission FP7
- 621394 / European Commission FP7
Conflict of Interest Statement
References
- Nagy A, Murray JK, Dyson S. Elimination from elite endurance rides in nine countries: A preliminary study.. Equine Vet J 2010;42:637–43.
- Legg KA, Weston J, Gee EK, Bolwell CF, Bridges JP, Rogers CW. Characteristics of Endurance Competitions and Risk Factors for Elimination in New Zealand during Six Seasons of Competition (2010/11–2015/16).. Animals 2019;6:611.
- Poole DC, Erickson HH. Cardiovascular function and oxygen transport: Responses to exercise and training.. In: Hinchcliff KW, Geor RJ, Kaneps AJ, editors. Equine Exercise Physiology: The Science of Exercise in the Athletic Horse. Philadelphia: Saunders Elsevier; 2008. p. 212–45.
- Mckeever KH. Body fluids and electrolytes: Responses to exercise and training.. In: Hinchcliff KW, Geor RJ, Kaneps AJ, editors. Equine Exercise Physiology: The Science of Exercise in the Athletic Horse. Philadelphia: Saunders Elsevier; 2008. p. 328–49.
- Lindiger MI, Ecker GL. Ion and water losses from body fluids during a 163 km endurance ride.. Equine Vet J 1995;18:314–22.
- Kingston JK, Bayly WM. Effect of Exercise on Acid-Base Status of Horses.. Vet Clin North Am Equine Pract 1998;14:61–73.
- Foreman JH. The Exhausted Horse Syndrome.. Vet Clin North Am Equine Pract 1998;14(1):205–19.
- Clayton HM. Conditioning Sport Horses.. Mason: Sport Horse Publications; 1991.
- Snow DH, Valberg SJ. Muscle anatomy, physiology and adaptations to exercise and training.. In: Hodgson DH, Rose RJ, editors. The Athletic horse: Principles and Practice of Equine Sports Medicine. Philadelphia: WB Saunders Co.; 1994. p. 145–79.
- Aebersold R, Mann M. Mass spectrometry-based proteomics.. Nature 2003;422(6928):198–207.
- Lee EC, Fragala MS, Kavouras SA, Queen RM, Pryor JL, Casa DJ. Biomarkers in Sports and Exercise: Tracking Health, Performance, and Recovery in Athletes.. J Strength Cond Res 2017;31(10):2920–37.
- Ichibangase T, Imai K. Application of fluorogenic derivatization-liquid chromatography-tandem mass spectrometric proteome method to skeletal muscle proteins in fast Thoroughbred horses.. J Proteome Res 2009;8(4):2129–34.
- Bouwman FG, van Ginneken MM, Noben JP, Royackers E, de Graaf-Roelfsema E, Wijnberg ID, van der Kolk JH, Mariman ECM, van Breda E. Differential expression of equine muscle biopsy proteins during normal training and intensified training in young standardbred horses using proteomics technology.. Comp Biochem Physiol Part D Genom Proteomics 2010;5:55–64.
- Scoppetta F, Tartaglia M, Renzone G, Avellini L, Gaiti A, Scaloni A, Chiaradia E. Plasma protein changes in horse after prolonged physical exercise: a proteomic study.. J Proteomics 2012;75(14):4494–504.
- Johansson L, Ringmark S, Bergquist J, Skiöldebrand, Widgren A, Jansson A. Proteomics perspective on 2 years of high-intensity training in horses: a pilot study.. Sci Rep 2024;14:23684.
- Horvatić A, Guillemin N, Kaab H, McKeegan D, O’Reilly E, Bain M, Kuleš J, Eckersall PD. Quantitative proteomics using tandem mass tags in relation to the acute phase protein response in chicken challenged with Escherichia coli lipopolysaccharide endotoxin.. J Proteomics 2019;192:64–77.
- Eckersall PD, Duthie S, Safi S, Moffatt D, Horadagoda NU, Doyle S, Parton R, Bennett D, Fitzpatrick JL. An automated biochemical assay for haptoglobin: Prevention of interference from albumin.. Comp Haematol Int 1999;9:117–24.
- Brady N, O’Reilly EL, McComb C, Macrae AI, Eckersall PD. An immunoturbidimetric assay for bovine haptoglobin.. Comp Clin Pathol 2019;28(1):21–7.
- Kuleš J, Bilić P, Beer Ljubić B, Gotić J, Crnogaj M, Brkljačić M, Mrljak V. Glomerular and tubular kidney damage markers in canine babesiosis caused by Babesia canis.. Ticks Tick Borne Dis 2018;9(6):1508–17.
- Team RC. R: a language and environment for statistical computing.. Vienna: R Foundation for Statistical Computing; 2020. Available from: https://www.R-project.org/.
- Wickham H. ggplot2 Elegant Graphics for Data Analysis.. New York: Springer Verlag; 2016.
- Yu G, Wang L-G, Han Y, He Q-Y. clusterProfiler: an R Package for Comparing Biological Themes Among Gene Clusters.. OMICS 2012;16(5):284–7.
- Yu G, He Q-Y. ReactomePA: an R/Bioconductor package for reactome pathway analysis and visualization.. Mol Biosyst 2016;12(2):477–9.
- Mach N, Ramayo-Caldas Y, Clark A, Moroldo M, Robert C, Barrey E, López JM, Le Moyec L. Understanding the response to endurance exercise using a systems biology approach: combining blood metabolomics, transcriptomics and miRNomics in horses.. BMC Genomics 2017;18:187.
- Qu JC, Ko W, Tso P, Bhargava A. Apolipoprotein A-IV: A Multifunctional Protein Involved in Protection against Atherosclerosis and Diabetes.. Cells 2019;8(4):319.
- Spaulding HL, Al E, Saijo F, Turnage RH, Alexander JS, Aw TY, Kalogeris TJ. Apolipoprotein A-IV attenuates oxidant-induced apoptosis in mitotic competent, undifferentiated cells by modulating intracellular glutathione redox balance.. Am J Physiol Cell Physiol 2006;290(1):95–103.
- Williams CA, Kronfeld DS, Hess TM, Saker KE, Waldron JE, Crandell KM, Harris PA. Comparison of oxidative stress and antioxidant status in endurance horses in three 80-km races.. Equine Comp Exerc Physiol 2005;2(03):153–7.
- Brkljača Bottegaro N, Gotić J, Šuran J, Brozić D, Klobučar K, Bojanić K, Vrbanac Z. Effect of prolonged submaximal exercise on serum oxidative stress biomarkers (d-ROMs, MDA, BAP) and oxidative stress index in endurance horses.. BMC Vet Res 2018;14(1):216.
- Goldberg IJ, Scheraldi CA, Yacoub LK, Saxena U, Bisagaier CL. Lipoprotein ApoC-II Activation of Lipoprotein Lipase.. J Biol Chem 1990;265(8):4266–72.
- Kersten S. Physiological regulation of lipoprotein lipase.. Biochim Biophys Acta 2014;1841(7):919–33.
- Petibois C, Cazorla G, Poortmans J-R, Deleris G. Biochemical aspects of overtraining in endurance sports: a review.. Sports Med 2002;32:867–78.
- Klein L, Miller TD, Radam TE, O'brien T, Nguyen TT, Kottke BA. Acute physical exercise alters apolipoprotein E and C-III concentrations of apo E-rich very low density lipoprotein fraction.. Atherosclerosis 1992;97(1):37–51.
- Melanson EL, Maclean PS, Hill JO. Exercise improves fat metabolism in muscle but does not increase 24-h fat oxidation.. Exerc Sport Sci Rev 2009;37(2):93–101.
- Borén J, Packard CJ, Taskinen MR. The Roles of ApoC-III on the Metabolism of Triglyceride-Rich Lipoproteins in Humans.. Front Endocrinol 2020;11:474.
- Amaya-Montoya M, Pinzón-Cortés JA, Silva-Bermúdez LS, Ruiz-Manco D, Pérez-Matos MC, Jiménez-Mora MA, Mendivil CO. ApoE and ApoC-III-defined HDL subtypes: a descriptive study of their lecithin cholesterol acyl transferase and cholesteryl ester transfer protein content and activity.. Lipids Health Dis 2020;19(1):106.
- Lawrence L. Nutrient Needs of Performance Horses.. Revista Brasileira de Zootecnia 2008;37:206–10.
- Harris P. Feeding management of elite endurance horses.. Vet Clin North Am Equine Pract 2009;25(1):137–53.
- Myćka G, Ropka-Molik K, Cywińska A, Szmatoła T, Stefaniuk-Szmukier M. Molecular insights into the lipid-carbohydrates metabolism switch under the endurance effort in Arabian horses.. Equine Vet J 2023.
- Smith SM, Melrose J. A Retrospective Analysis of the Cartilage Kunitz Protease Inhibitory Proteins Identifies These as Members of the Inter-α-Trypsin Inhibitor Superfamily with Potential Roles in the Protection of the Articulatory Surface.. Int J Mol Sci 2019;20(3):497.
- Te Moller NCR, van Weeren PR. How exercise influences equine joint homeostasis.. Vet J 2017;222:60–7.
- Murray RC, Birch HL, Lakhani K, Goodship AE. Biochemical composition of equine carpal articular cartilage is influenced by short-term exercise in a site-specific manner.. Osteoarthritis Cartilage 2001;9(7):625–32.
- Mihelić K, Vrbanac Z, Bojanić K, Kostanjšak T, Beer Ljubić B, Gotić J, Vnuk D, Brkljača BN. Changes in Acute Phase Response Biomarkers in Racing Endurance Horses.. Animals 2022;12(21):2993.
- Bertaggia E, Scabia G, Dalise S, Verso FL, Santini F, Vitti P, Chisari C, Sandri M, Maffei M. Haptoglobin Is Required to Prevent Oxidative Stress and Muscle Atrophy.. PLOS ONE 2014;9(6):e100745.
- Lippi G, Sanchis-Gomar F. Epidemiological, biological and clinical update on exercise induced hemolysis.. Ann Transl Med 2019;7(12):270.
- Kobayashi Y, Nakatsuji A, Aoi W, Wada S, Kuwahata M, Kido Y. Intense Exercise Increases Protein Oxidation in Spleen and Liver of Mice.. Nutr Metab Insights 2014;7:1–6.
- Rifkind JM, Mohanty JG, Nagababu E. The pathophysiology of extracellular hemoglobin associated with enhanced oxidative reactions.. Front Physiol 2015;5:500.
- Bicho MC, da Silva AP, Medeiros R, Bicho M. The Role of Haptoglobin and Its Genetic Polymorphism in Cancer: A Review.. Acute Phase Proteins InTech 2013.
- Sakata S, Yoshioka N, Atassi MZ. Human haptoglobin binds to human myoglobin.. Biochim Biophys Acta 1986;873(2):312–5.
- Schott HC, Marlin RJ, Geor RJ, Holbrooks TC, Deaton CM, Vincent T, Dacrey K, Schroter RC, Jose-Cunillera E, Cornelisse CJ. Changes in selected physiological and laboratory measurements in elite horses competing in a 160 km endurance ride.. Equine Vet J 2006;36:37–42.
- Klobučar K, Vrbanac Z, Gotić J, Bojanić K, Bureš T, Brkljača BN. Changes in biochemical parameters in horses during 40 km and 80 km endurance races.. Acta Vet-Beogr 2019;69(1):73–87.
- Pellegrini Masini A, Tedeschi D, Baragli P, Sighieri C, Lubas G. Exercise-induced intravascular haemolysis in standardbred horses.. Comp Clin Path 2003;12:45–8.
- Cywińska A, Szarska E, Gorecka R, Witkowski L, Hecold M, Bereznowski A, Schollenberger A, Winnicka A. Acute phase protein concentrations after limited distance and long distance endurance rides in horses.. Res Vet Sci 2012;93(3):1402–6.
- Kawabata H. Transferrin and transferrin receptors update.. Free Radic Biol Med 2019;133:46–54.
- Schumacher YO, Schmid A, König D, Berg A. Effects of exercise on soluble transferrin receptor and other variables of the iron status.. Br J Sports Med 2002;36(3):195–9.
- Smith JE. Effects of strenuous exercise on haemostasis.. Br J Sports Med 2003;37(5):433–5.
- Ferguson EW, Bernier LL, Banta GR, Yu-Yahiro J, Schoomaker EB. Effects of exercise and conditioning on clotting and fibrinolytic activity in men.. J Appl Physiol 1987;62(4):1416–21.
- Paltrinieri S, Meazza C, Giordano A, Tunesi C. Validation of thromboelastometry in horses.. Vet Clin Pathol 2008;37(3):277–85.
- Pilecki B, Holm AT, Schlosser A, Moeller JB, Wohl AP, Zuk AV, Heumüller SE, Wallis R, Moestrup SK, Sengle G, Holmskov U, Sorensen GL. Characterization of microfibrillar-associated protein 4 (MFAP4) as a tropoelastin- and fibrillin-binding protein involved in elastic fiber formation.. J Biol Chem 2016;291(3):1103–14.
- Mölleken C, Poschmann G, Bonella F, Costabel U, Sitek B, Stühler K, Meyer HE, Schmiegel WH, Marcussen N, Helmer M, Nielsen O, Hansen S, Schlosser A, Holmskov U, Lykke SG. MFAP4: a candidate biomarker for hepatic and pulmonary fibrosis?. Sarcoidosis, Vasc Diffuse Lung Dis 2016;33(1):41–50.
- Johansson SL, Roberts NB, Schlosser A, Andersen CB, Carlsen J, Wulf-Johansson H, Sækmose SG, Titlestad IL, Tornoe I, Miller B, Tal-Singer R, Holmskov U, Vestbo J, Lykke SG. Microfibrillar-associated protein 4: a potential biomarker of chronic obstructive pulmonary disease.. Respir Med 2014;108(9):1336–44.
- Davalieva K, Kostovska IM, Kiprijanovska S, Markoska K, Kubelka-Sabit K, Filipovski V, Stavridis S, Stankov O, Komina S, Petrusevska G, Polenakovic M. Proteomics analysis of malignant and benign prostate tissue by 2D DIGE/MS reveals new insights into proteins involved in prostate cancer.. Prostate 2015;75(14):1586–600.
- Zhao H, Sun Q, Li L, Zhou J, Zhang C, Hu T, Zhou X, Zhang L, Wang B, Li B, Zhu T, Li H. High expression levels of AGGF1 and MFAP4 predict primary platinum-based chemoresistance and are associated with adverse prognosis in patients with serous ovarian cancer.. J Cancer 2019;10(2):397–407.
- Yang J, Song H, Chen L, Cao K, Zhang Y, Li Y, Hao X. Integrated analysis of microfibrillar-associated proteins reveals MFAP4 as a novel biomarker in human cancers.. Epigenomics 2019;11(1):15–21.
- Bilić P, Guillemin N, Kovačević A, Beer Ljubić B, Jović I, Galan A, Eckersall PD, Burchmore R, Mrljak V. Serum proteome profiling in canine idiopathic dilated cardiomyopathy using TMT-based quantitative proteomics approach.. J Proteomics 2018;179:110–21.
- Sækmose SG, Holst R, Lottenburger T, Ytting H, Nielsen HJ, Junker P, Schlosser A, Sorensen GL. Circadian, Week-to-Week, and Physical Exercise-Induced Variation of Serum Microfibrillar-Associated Protein 4.. Biomarker Insights 2021;16:11772719211016360.