Serum protein expression in Equine Glandular Gastric Disease (EGGD) induced by phenylbutazone

Abstract

Equine Glandular Gastric Disease (EGGD) is a common disease in sport horses. This disease might be associated with usage of nonsteroidal anti-inflammatory drugs (NSAIDs) for treating inflammatory diseases. Although gastroscopy has been an effective method for diagnosis, but a less invasive, and inexpensive method is preferred. This study used proteomic technology to identify candidate serum proteins that might be used as markers of NSAIDs induced EGGD. Five Thoroughbred horses were given high doses of NSAID, phenylbutazone to treat lameness. The experiment was divided into three periods: (i) Pre-EGGD period, (ii) during EGGD period, and (iii) Post-EGGD period. Gastroscopy were used to diagnose EGGD, serum was collected to perform gel electrophoresis (1D SDS-PAGE) and mass spectrometry (LC-MS) in order to identify serum proteins in each group. The candidate serum proteins were computationally predicted for the interaction between phenylbutazone and proteins, tissue specific expression, and association to gastric ulceration. After EGGD induction, all horses showed clinical signs of colic with marked congestion and erosion appearing in the mucosa of the glandular stomach whereas no change was observed in the mucosa of non-glandular stomach. Our proteomic results identified 14 proteins that might be used as EGGD markers. These proteins were highly expressed in the glandular stomach and some proteins were associated with phenylbutazone or ulcer development. However, confirmation of these candidate marker proteins is required with specific antibodies in the larger horse population before they can be considered for application in the field.

Equine Gastric Ulcer Syndrome: An Update on Current Knowledge

“…Many clinical signs have been ascribed to gastric mucosal disease over the years, but surprisingly few studies have investigated these on a large scale. Proposed clinical signs, discussed below, include colic [ 4 , 39 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 ], weight loss or poor body condition [ 4 , 42 , 51 , 52 , 56 , 57 ], poor coat condition [ 39 ], reduced appetite [ 4 , 39 , 51 , 52 , 56 , 58 ], diarrhea [ 51 ], bruxism [ 51 , 52 ], flehmen response [ 37 ], behavioral changes [ 43 , 52 , 56 , 57 , 59 ] and poor performance [ 14 , 37 , 39 , 52 , 56 , 57 , 60 , 61 , 62 ]. In interpreting the above, it is important to distinguish between ESGD and EGGD as the majority of literature is weighted towards ESGD, and a recent study found no associations between owner-reported clinical signs and EGGD [ 63 ].…”

“…Furthermore, several population-based studies have failed to find NSAID use associated with increased risk of EGGD [ 28 , 30 , 90 ]. At an experimental level, multiple studies have shown the ulcerogenic capacity of NSAIDs at high doses in horses [ 50 , 91 , 92 , 93 , 94 , 95 , 96 , 97 ]. Most of these studies, however, use NSAID doses in excess of what is typically recommended for clinical use [ 98 ].…”

“…Similarly, studies show that hematology and biochemistry or inflammatory markers, such as serum amyloid A (SAA), have little clinical use in the diagnosis of, or screening for either ESGD or EGGD [ 126 , 127 ]. Research into novel serum and salivary testing for detecting ESGD and EGGD has been undertaken with some differences demonstrated between diseased and control populations, although a large degree of overlap exists between populations [ 50 , 89 , 126 , 128 , 129 ]. The clinical utility of these tests is very limited at this point in time.…”

Evaluation of gastrointestinal tract lesions and serum malondialdehyde levels after repeated oral administration of phenylbutazone in horses

“…Proteomics can be a helpful tool in early diagnosis of various diseases, including EGUS. Ten protein markers for equine squamous gastric disease (ESGD) and 14 for EGGD have been successfully identified through proteomics (Tesena et al 2019). Some of these peptides are biomarkers of oxidative stress, and oxidative stress seems to play an important role in various equine gastrointestinal diseases, including colic (Hajimohammadi et al 2023), EGGD (Banse and Andrews 2019), EGUS (Shawaf et al 2020), and colitis (El-Ashker et al 2012), by causing injury to the gastrointestinal mucosa.…”