A comprehensive and comparative proteomic analysis of horse serum proteins in colitis

Equine Vet J. 2022 Nov;54(6):1039-1046. doi: 10.1111/evj.13554. Epub 2022 Jan 18.

Abstract

Background: Equine colitis is a diarrhoeal disease caused by inflammation of the large bowel and can potentially be life-threatening due to its rapid progression. Pathogenesis is multifactorial and pathophysiology is highly complicated, therefore, reliable diagnostic biomarkers are needed in the veterinary field.

Objective: Serum is one of the most commonly used diagnostic tools in equine clinical investigation. To discover diagnostic or prognostic protein markers for colitis in horse serum, comprehensive and comparative proteomic analysis was conducted using liquid chromatography-tandem mass spectrometry (LC-MS/MS).

Study design: Case-control study.

Methods: Serum samples were collected from 36 healthy Thoroughbreds and 12 Thoroughbreds with colitis. Serum from each horse suffering from colitis was collected daily until death or recovery. Collected sera were digested with trypsin. Peptides obtained from serum proteins were measured by Q-Exactive HF Orbitrap mass spectrometer. The identification and quantification of peptides were performed using Proteome Discoverer version 2.2.

Results: On day 1 of treatment, eight proteins in the colitis group were upregulated (P < .05, more than a twofold change) compared with the healthy group. Among the eight proteins, biliverdin reductase B was significantly upregulated (P < .05) in the non-survivor group (n = 5) compared with the survivor group (n = 7). On the last day of the treatment, haemoglobin subunit alpha, clusterin, glyceraldehyde-3-phosphate dehydrogenase, lipopolysaccharide-binding protein, and biliverdin reductase B showed significant increases (P < .05) in the non-survivor group.

Main limitations: The number of the identified proteins is limited due to the existence of abundant proteins.

Conclusions: Measuring the changes of these proteins together may enable a potential prognosis or early diagnosis of horses suffering from colitis.

Keywords: LC-MS/MS; colitis; horse; proteome; serum.

MeSH terms

  • Animals
  • Biomarkers
  • Blood Proteins / analysis
  • Case-Control Studies
  • Chromatography, Liquid / methods
  • Chromatography, Liquid / veterinary
  • Clusterin
  • Colitis* / veterinary
  • Hemoglobin Subunits / analysis
  • Horse Diseases* / diagnosis
  • Horses
  • Peptides
  • Proteome / analysis
  • Proteomics / methods
  • Tandem Mass Spectrometry / veterinary
  • Trypsin

Substances

  • Biomarkers
  • Blood Proteins
  • Clusterin
  • Hemoglobin Subunits
  • Peptides
  • Proteome
  • Trypsin