Quantitative proteomics unveils potential plasma biomarkers and provides insights into the pathophysiological mechanisms underlying equine metabolic syndrome.
Abstract: Equine Metabolic Syndrome (EMS) is a multifactorial endocrine disorder characterized by obesity, insulin dysregulation (ID), and an increase in the risk of laminitis, a painful condition that can lead to euthanasia in severe cases. Diagnosing EMS is challenging and often relies on clinical history including obesity, difficulty in losing weight, and recurring episodes of laminitis. The gold standard for laboratory support of an EMS diagnosis is the identification of ID, with basal insulin being the simplest and most accessible method, especially in a field setting. However, various factors such as diet, age, stress, season, medications administered, and testing protocols can influence results. Dynamic tests like the oral sugar test (OST) are preferred but present limitations due to low sensitivity and poor repeatability. These diagnostic challenges make EMS difficult to detect in veterinary medicine highlighting the need for an effective method of the early detection of EMS to prevent laminitis and its associated complications. Mass spectrometry-based proteomics represents a powerful tool to identify biomarkers and explore molecular pathways related to the underlying pathology. In the current study we established an integrated proteomics pipeline to identify plasma biomarkers for EMS diagnosis. We compared plasma proteomes from healthy horses, non-ID obese horses and animals diagnosed with EMS. This comparison revealed 76 proteins with significant changes (1% FDR) between groups. Our study demonstrates that the complement system, the coagulation cascade and extracellular matrix remodelling pathways are altered in EMS. These findings offer new insights into the molecular basis of the development of EMS and led to the nomination of several proteins as potential biomarkers for its early detection. The online version contains supplementary material available at 10.1186/s12917-025-04879-6.
Publication Date: 2025-07-02 PubMed ID: 40604814PubMed Central: PMC12217909DOI: 10.1186/s12917-025-04879-6Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research paper discusses the exploration of Equine Metabolic Syndrome (EMS) through methods in proteomics, in particular, identifying potential protein biomarkers in plasma that could aid early diagnosis of EMS and provide a better understanding of the disorder’s underlying molecular pathways.
Understanding Equine Metabolic Syndrome
- The focal point of this research is a condition known as Equine Metabolic Syndrome (EMS), a complex hormonal disorder in horses significantly marked by obesity, erratic insulin regulation, and a higher risk of laminitis, a painful condition that often leads to euthanasia.
- Due to factors such as diet, age, season, stress, and medications, diagnosing EMS poses challenges, particularly given that its diagnosis is frequently based on clinical history and observed symptoms such as recurring laminitis and challenges with weight loss.
Role of Proteomics in EMS Diagnosis
- The research acknowledges the limitations of current diagnostic methods, such as blood tests for basal insulin and oral sugar tests (OST). They point to mass spectrometry-based proteomics as a promising tool for identification of biomarkers and exploring disease pathology.
- The authors leveraged proteomics, a branch of biology concerned with the study of proteomes (sets of proteins), to establish an integrated pipeline for identifying potential blood plasma biomarkers that could support early EMS detection.
Comprehensive Proteomic Study and Findings
- The authors conducted a comprehensive study comparing the plasma proteomes from healthy horses, non-insulin dysregulated obese horses, and horses already diagnosed with EMS.
- This led to the identification of 76 proteins that showed significant changes between groups, denoting some potential biomarkers for EMS diagnosis.
- Key pathways that showed alteration in EMS cases included the complement system, the coagulation cascade, and extracellular matrix remodelling. This can guide further understanding into the molecular mechanisms behind EMS development.
Conclusions and Future Applications
- This research offers in-depth insights into the molecular processes of EMS and presents several proteins as potential biomarkers for early detection.
- Such findings could serve as a stepping stone for further exploration and development of more effective diagnostic methods and interventions for EMS in equine veterinary medicine.
Cite This Article
APA
Espinosa-López EM, Ortiz-Guisado B, Diez de Castro E, Durham A, Aguilera-Tejero E, Gómez-Baena G.
(2025).
Quantitative proteomics unveils potential plasma biomarkers and provides insights into the pathophysiological mechanisms underlying equine metabolic syndrome.
BMC Vet Res, 21(1), 425.
https://doi.org/10.1186/s12917-025-04879-6 Publication
Researcher Affiliations
- Department of Biochemistry and Molecular Biology, Faculty of Veterinary Sciences, University of Córdoba, Córdoba, Spain.
- Department of Biochemistry and Molecular Biology, Faculty of Veterinary Sciences, University of Córdoba, Córdoba, Spain.
- Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, University of Córdoba, Córdoba, Spain.
- Liphook Equine Hospital, Liphook, UK.
- Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, University of Córdoba, Córdoba, Spain.
- Department of Biochemistry and Molecular Biology, Faculty of Veterinary Sciences, University of Córdoba, Córdoba, Spain. v52gobag@uco.es.
Conflict of Interest Statement
Declarations. Ethics approval and consent to participate: The study protocol received approval from the Institutional Committee at the Veterinary Hospital of the University of Córdoba (Spain) and the University of Extremadura (Spain). Written informed consent was obtained from the owners of all horses enrolled in the study. All diagnostic procedures were clinically indicated, aimed at benefiting the animals, and were conducted in compliance with the highest standards of veterinary practice. The authors confirm that all methods were carried out in accordance with relevant guidelines and veterinary regulations. Consent to publish: Not applicable. Competing interests: The authors declare no competing interests.
References
This article includes 77 references
- Durham AE. ECEIM consensus statement on equine metabolic syndrome. J Vet Intern Med 2019;33:335–49.
- Hunt RJ. A retrospective evaluation of laminitis in horses. Equine Vet J 1993;25:61–4.
- Pollard D, Wylie CE, Newton JR, Verheyen KLP. Factors associated with euthanasia in horses and ponies enrolled in a laminitis cohort study in great Britain. Prev Vet Med 2020;174:104833.
- Hart KA. Effect of age, season, body condition, and endocrine status on serum free cortisol fraction and insulin concentration in horses. J Vet Intern Med 2016;30:653–63.
- Macon EL, Harris P, Barker VD, Adams AA. Seasonal insulin responses to the oral sugar test in healthy and insulin dysregulated horses. J Equine Vet Sci 2022;113:103945.
- Jacquay ET, Harris PA, Adams AA. The impact of short-term transportation stress on insulin and oral sugar responses in insulin dysregulated and non-insulin dysregulated horses. Equine Vet J 2024.
- McFarlane D. Diagnostic testing for equine endocrine diseases: confirmation versus confusion. Vet Clin North Am Equine Pract 2019;35:327–38.
- Banse HE, McCann J, Yang F, Wagg C, McFarlane D. Comparison of two methods for measurement of equine insulin. J Vet Diagn Invest 2014;26:527–30.
- Borer-Weir KE, Bailey SR, Menzies-Gow NJ, Harris PA, Elliott J. Evaluation of a commercially available radioimmunoassay and species-specific ELISAs for measurement of high concentrations of insulin in equine serum. Am J Vet Res 2012;73:1596–602.
- Tinworth KD. Evaluation of commercially available assays for the measurement of equine insulin. Domest Anim Endocrinol 2011;41:81–90.
- de Laat MA. Carbohydrate pellets to assess insulin dysregulation in horses. J Vet Intern Med 2022.
- de Laat MA, McGree JM, Sillence MN. Equine hyperinsulinemia: investigation of the enteroinsular axis during insulin dysregulation. Am J Physiol Endocrinol Metab 2016;310:E61–72.
- Karikoski NP, Box JR, Mykkanen AK, Kotiranta VV, Raekallio MR. Variation in insulin response to oral sugar test in a cohort of horses throughout the year and evaluation of risk factors for insulin dysregulation. Equine Vet J 2022;54:905–13.
- Hopster K, Driessen B. Pharmacology of the equine foot: medical pain management for laminitis. Vet Clin North Am Equine Pract 2021;37:549–61.
- Henneke DR, Potter GD, Kreider JL, Yeates BF. Relationship between condition score, physical measurements and body fat percentage in mares. Equine Vet J 1983;15:371–2.
- Fitzgerald DM, Anderson ST, Sillence MN, de Laat MA. The Cresty neck score is an independent predictor of insulin dysregulation in ponies. PLoS ONE 2019;14:e0220203.
- Macon EL, Harris P, Partridge E, Barker VD, Adams A. Effect of dose and fasting on oral sugar test responses in insulin dysregulated horses. J Equine Veterinary Sci 2021;107:103770.
- Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680–5.
- Gómez-Baena G. Molecular complexity of the major urinary protein system of the Norway rat, Rattus norvegicus. Sci Rep 2019;9:10757.
- Gómez-Baena G. Unraveling female communication through scent marks in the Norway rat. Proc Natl Acad Sci U S A 2023;120:e2300794120.
- Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248–54.
- Gómez-Baena G. Quantitative proteomics of cerebrospinal fluid in paediatric Pneumococcal meningitis. Sci Rep 2017;7:7042.
- Team RC. R: A Language and Environment for Statistical Computing_. (2024).
- Larsson J. eulerr: Area-proportional Euler and Venn Diagrams with Ellipses. (2024).
- Kolde R. pheatmap: Pretty Heatmaps. (2019).
- Le S, Josse J, Husson F. FactoMineR: an R package for multivariate analysis. J Stat Softw 2008;25:1–18.
- Kassambara A, Mundt F. factoextra: Extract and Visualize the Results of Multivariate Data Analyses. (2020).
- Kolberg L. g:Profiler-interoperable web service for functional enrichment analysis and gene identifier mapping (2023 update). Nucleic Acids Res 2023;51:W207–12.
- Anderson FDMST, Sillence MN, de Laat MA. The Cresty neck score is an independent predictor of insulin dysregulation in ponies. PLoS ONE 2019;14:e0220203.
- Delarocque J. Development of a web app to convert blood insulin concentrations among various immunoassays used in horses. Anim (Basel) 13 (2023).
- Knowles EJ, Harris PA, Elliott J, Chang YM. Menzies-Gow, factors associated with insulin responses to oral sugars in a mixed-breed cohort of ponies. Equine Vet J 2024;56:253–63.
- Wang H. MultiPro: DDA-PASEF and diapasef acquired cell line proteomic datasets with deliberate batch effects. Sci Data 2023;10:858.
- Johansson L. A proteomics perspective on 2 years of high-intensity training in horses: a pilot study. Sci Rep 2024;14:23684.
- Kopp A, Hebecker M, Svobodova E, Jozsi M. Factor h: a complement regulator in health and disease, and a mediator of cellular interactions. Biomolecules 2012;2:46–75.
- Hertle E, Stehouwer CD, van Greevenbroek MM. The complement system in human cardiometabolic disease. Mol Immunol 2014;61:135–48.
- Muscari A. Serum C3 is a stronger inflammatory marker of insulin resistance than C-reactive protein, leukocyte count, and erythrocyte sedimentation rate: comparison study in an elderly population. Diabetes Care 2007;30:2362–8.
- Liu Z. Elevated serum complement factors 3 and 4 are strong inflammatory markers of the metabolic syndrome development: a longitudinal cohort study. Sci Rep 2016;6:18713.
- Nieuwdorp M, Stroes ES, Meijers JC, Buller H. Hypercoagulability in the metabolic syndrome. Curr Opin Pharmacol 2005;5:155–9.
- Zak A. Effects of equine metabolic syndrome on inflammation and acute-phase markers in horses. Domest Anim Endocrinol 2020;72:106448.
- Adams AA. Effect of body condition, body weight and adiposity on inflammatory cytokine responses in old horses. Vet Immunol Immunopathol 2009;127:286–94.
- Elzinga S, Wood P, Adams AA. Plasma lipidomic and inflammatory cytokine profiles of horses with equine metabolic syndrome. J Equine Veterinary Sci 2016;40:49–55.
- Holbrook TC, Tipton T, McFarlane D. Neutrophil and cytokine dysregulation in hyperinsulinemic obese horses. Vet Immunol Immunopathol 2012;145:283–9.
- Ragno VM. Morphometric, metabolic, and inflammatory markers across a cohort of client-owned horses and ponies on the insulin dysregulation spectrum. J Equine Vet Sci 2021;105:103715.
- Suagee JK, Splan RK, Swyers KL, Geor RJ, Corl BA. Effects of high-sugar and high-starch diets on postprandial inflammatory protein concentration in horses. J Equine Veterinary Med 2015;35:191–7.
- Vick MM. Relationships among inflammatory cytokines, obesity, and insulin sensitivity in the horse. J Anim Sci 2007;85:1144–55.
- Lovett AL, Gilliam LL, Sykes BW, McFarlane D. Thromboelastography in obese horses with insulin dysregulation compared to healthy controls. J Vet Intern Med 2021;36:1131–8.
- de Laat-Kremers R. High alpha-2-macroglobulin levels are a risk factor for cardiovascular disease events: A Moli-sani cohort study. Thromb Res 2024;234:94–100.
- Campolo A. Differential proteomic expression of equine cardiac and lamellar tissue during Insulin-Induced laminitis. Front Vet Sci 2020;7:308.
- Oikonomopoulou K, Ricklin D, Ward PA, Lambris JD. Interactions between coagulation and complement–their role in inflammation. Semin Immunopathol 2012;34:151–65.
- Bekassy Z, Lopatko Fagerstrom I, Bader M, Karpman D. Crosstalk between the renin-angiotensin, complement and kallikrein-kinin systems in inflammation. Nat Rev Immunol 2022;22:411–28.
- Lopatko Fagerstrom I. Blockade of the kallikrein-kinin system reduces endothelial complement activation in vascular inflammation. EBioMedicine 2019;47:319–28.
- Leung LLK, Morser J. Carboxypeptidase B2 and carboxypeptidase N in the crosstalk between coagulation, thrombosis, inflammation, and innate immunity. J Thromb Haemost 2018.
- Jones AL, Hulett MD, Parish CR. Histidine-rich glycoprotein: A novel adaptor protein in plasma that modulates the immune, vascular and coagulation systems. Immunol Cell Biol 2005;83:106–18.
- Bourebaba L, Marycz K. Pathophysiological implication of Fetuin-A glycoprotein in the development of metabolic disorders: A concise review. J Clin Med 8 (2019).
- Pan X. Fetuin-A in metabolic syndrome: A systematic review and meta-analysis. PLoS ONE 2020;15:e0229776.
- Chekol Abebe E. The structure, biosynthesis, and biological roles of fetuin-A: A review. Front Cell Dev Biol 2022;10:945287.
- Reinehr T, Roth CL. Fetuin-A and its relation to metabolic syndrome and fatty liver disease in obese children before and after weight loss. J Clin Endocrinol Metab 2008;93:4479–85.
- Wang Y, Koh WP, Jensen MK, Yuan JM, Pan A. Plasma Fetuin-A levels and risk of type 2 diabetes mellitus in A Chinese population: A nested Case-Control study. Diabetes Metab J 2019;43:474–86.
- Pal D. Fetuin-A acts as an endogenous ligand of TLR4 to promote lipid-induced insulin resistance. Nat Med 2012;18:1279–85.
- Stefan N. Alpha2-Heremans-Schmid glycoprotein/fetuin-A is associated with insulin resistance and fat accumulation in the liver in humans. Diabetes Care 2006;29:853–7.
- Meex RC. Fetuin B is a secreted hepatocyte factor linking steatosis to impaired glucose metabolism. Cell Metabol 2015;22:1078–89.
- Xue S. Serum Fetuin-B Levels Are Elevated in Women with Metabolic Syndrome and Associated with Increased Oxidative Stress. 2021, 6657658 (2021).
- Pasmans K. Fetuin B in white adipose tissue induces inflammation and is associated with peripheral insulin resistance in mice and humans. Obes (Silver Spring) 2023.
- Mokou M. Elevated Circulating Fetuin-B Levels Are Associated with Insulin Resistance and Reduced by GLP-1RA in Newly Diagnosed PCOS Women. 2020, 2483435 (2020).
- Xia X. Association of serum fetuin-B with insulin resistance and pre-diabetes in young Chinese women: evidence from a cross-sectional study and effect of liraglutide. PeerJ 9, e11869 (2021).
- Guzmán-Ruiz R. Adipose tissue depot-specific intracellular and extracellular cues contributing to insulin resistance in obese individuals. FASEB J 2020;34:7520–39.
- Brezillon S, Pietraszek K, Maquart FX, Wegrowski Y. Lumican effects in the control of tumour progression and their links with metalloproteinases and integrins. FEBS J 2013;280:2369–81.
- Strieder-Barboza C. Lumican modulates adipocyte function in obesity-associated type 2 diabetes. Adipocyte 2022;11:665–75.
- Wolff G. Diet-dependent function of the extracellular matrix proteoglycan lumican in obesity and glucose homeostasis. Mol Metab 2019;19:97–106.
- Gill SE, Parks WC. Metalloproteinases and their inhibitors: regulators of wound healing. Int J Biochem Cell Biol 2008;40:1334–47.
- Kyaw-Tanner M, Pollitt CC. Equine laminitis: increased transcription of matrix metalloproteinase-2 (MMP-2) occurs during the developmental phase. Equine Vet J 2004;36:221–5.
- Scholze A. Plasma concentrations of extracellular matrix protein fibulin-1 are related to cardiovascular risk markers in chronic kidney disease and diabetes. Cardiovasc Diabetol 2013;12:6.
- Cangemi C. Fibulin-1 is a marker for arterial extracellular matrix alterations in type 2 diabetes. Clin Chem 2011;57:1556–65.
- Ye JJ, Bian X, Lim J, Medzhitov R. Adiponectin and related C1q/TNF-related proteins bind selectively to anionic phospholipids and sphingolipids. Proc Natl Acad Sci U S A 2020;117:17381–8.
- Wooldridge AA. Evaluation of high-molecular weight adiponectin in horses. Am J Vet Res 2012;73:1230–40.
- Ruan W, Lai M. Insulin-like growth factor binding protein: a possible marker for the metabolic syndrome?. Acta Diabetol 2010;47:5–14.
- Carter RA, Geor RJ, Burton Staniar W, Cubitt TA, Harris PA. Apparent adiposity assessed by standardised scoring systems and morphometric measurements in horses and ponies. Vet J 2009;179:204–10.
Citations
This article has been cited 0 times.Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
