Novel link between inflammation and impaired glucose transport during equine insulin resistance.

Although insulin resistance (IR) has been increasingly recognized in horses, a clear understanding of its pathophysiology is lacking. The purpose of the present study was to determine the early pathologic changes in IR horses by characterizing alterations in proteins that play key roles in innate immunological responses and inflammatory pathways, and by identifying potential links with glucose transport and insulin signaling. Visceral (VIS) and subcutaneous (SC) adipose tissue and skeletal muscle (SM) biopsies were collected from horses, which were classified as insulin-sensitive (IS) or IR based on the results of an insulin-modified frequently sampled intravenous glucose tolerance test. Protein expression of Toll-like receptor 4 (TLR-4), suppressor of cytokine signaling 3 (SOCS-3) and tumor necrosis factor alpha (TNF-α) were quantified by Western blotting in VIS and SC adipose depots and SM, as well as insulin receptor substrate 1 (IRS-1). To better characterize the potential relationship between inflammation, IR and impaired glucose transport, we correlated active cell surface glucose transporter 4 (GLUT-4) content (measured by a cell surface biotinylated assay) with individual- and tissue-specific data related to inflammation. IR was associated with a significantly increased expression of TLR-4 and SOCS-3 in SM and VIS tissue, without a significant change in SC site. We also observed a significant increase in TNF-α in VIS, but not in SC, tissue of IR vs. IS horses. There was no difference in total content or serine phosphorylation of IRS-1 for any sampling site in IR compared to IS horses. We further observed a significant positive correlation between TLR-4 content and SOCS-3, as well as a significant negative correlation between SOCS-3 content and GLUT-4 trafficking. Taken together, the data suggested a pro-inflammatory state in SM and VIS, but not SC, adipose depot during compensated IR. In addition, SOCS-3 appears to be a novel link between inflammation and dysregulated glucose metabolism and insulin sensitivity during the early pathogenesis of insulin resistance.

Naturally occurring compensated insulin resistance selectively alters glucose transporters in visceral and subcutaneous adipose tissues without change in AS160 activation.

Although the importance of adipose tissue (AT) glucose transport in regulating whole-body insulin sensitivity is becoming increasingly evident and insulin resistance (IR) has been widely recognized, the underlying mechanisms of IR are still not well understood. The purpose of the present study was to determine the early pathological changes in glucose transport by characterizing the alterations in glucose transporters (GLUT) in multiple visceral and subcutaneous adipose depots in a large animal model of naturally occurring compensated IR. AT biopsies were collected from horses, which were classified as insulin-sensitive (IS) or compensated IR based on the results of an insulin-modified frequently sampled intravenous glucose tolerance test. Protein expression of GLUT4 (major isoform) and GLUT12 (one of the most recently discovered isoforms) were measured by Western blotting in multiple AT depots, as well as AS160 (a potential key player in GLUT trafficking pathway). Using a biotinylated bis-mannose photolabeled technique, active cell surface GLUT content was quantified. Omental AT had the highest total GLUT content compared to other sites during the IS state. IR was associated with a significantly reduced total GLUT4 content in omental AT, without a change in content in other visceral or subcutaneous adipose sites. In addition, active cell surface GLUT-4, but not -12, was significantly lower in AT of IR compared to IS horses, without change in AS160 phosphorylation between groups. Our data suggest that GLUT4, but not GLUT12, is a pathogenic factor in AT during naturally occurring compensated IR, despite normal AS160 activation.

Insulin resistance selectively alters cell-surface glucose transporters but not their total protein expression in equine skeletal muscle.

BACKGROUND: Insulin resistance (IR) has been widely recognized in humans, and more recently in horses, but its underlying mechanisms are still not well understood. The translocation of glucose transporter 4 (GLUT4) to the cell surface is the limiting step for glucose uptake in insulin-sensitive tissues. Although the downstream signaling pathways regulating GLUT translocation are not well defined, AS160 recently has emerged as a potential key component. In addition, the role of GLUT12, one of the most recently identified insulin-sensitive GLUTs, during IR is unknown. HYPOTHESIS/OBJECTIVES: We hypothesized that cell-surface GLUT will be decreased in muscle by an AS160-dependent pathway in horses with IR. ANIMALS: Insulin-sensitive (IS) or IR mares (n = 5/group). METHODS: Muscle biopsies were performed in mares classified as IS or IR based on results of an insulin-modified frequently sampled IV glucose tolerance test. By an exofacial bis-mannose photolabeled method, we specifically quantified active cell-surface GLUT4 and GLUT12 transporters. Total GLUT4 and GLUT12 and AS160 protein expression were measured by Western blots. RESULTS: IR decreased basal cell-surface GLUT4 expression (P= .027), but not GLUT12, by an AS160-independent pathway, without affecting total GLUT4 and GLUT12 content. Cell-surface GLUT4 was not further enhanced by insulin stimulation in either group. CONCLUSIONS AND CLINICAL IMPORTANCE: IR induced defects in the skeletal muscle glucose transport pathway by decreasing active cell-surface GLUT4.

Factors contributing to plasma TCO2 and acid-base state in Ontario Standardbred racehorses.

REASONS FOR PERFORMING STUDY: Standardbred and Thoroughbred racehorses around the world are tested for performance enhancing substances. Among these are blood alkalising substances that raise plasma pH and total carbon dioxide (TCO(2)) concentration. However, many horses have an elevated TCO(2) due to dietary, environmental and health concerns without having been administered an alkalising substance. OBJECTIVES: The purposes of this study were to determine the acid-base profile of a cross section of Standardbred horses in racing/race training in Ontario and the main independent variables that contributed to acid-base state. MATERIALS AND METHODS: On nonracing days, blood from 211 horses at rest, from 9 training facilities, was analysed within 30 min for plasma pH (7.406 ± 0.039; mean ± s.e.), PCO(2) (50.0 ± 3.4 mmHg), from which [HCO(3)(-)] (31.2 ± 2.8 mmol/l) and [TCO(2)] (33.1 ± 2.9 mmol/l; range 25.66-42.9) were calculated. From these, a subset of 161 horses had full data sets for plasma protein and strong ion concentrations. These data were further analysed by facility and level of TCO(2). Data on nutrition, training, racing and medications were also collected. RESULTS: There were significant differences amongst facilities with respect to plasma pH, TCO(2), strong ion difference ([SID]), PCO(2) and total weak acid concentration ([A(tot)]). Horses having the highest TCO(2) (37.0-42.9 mmol/l, n = 16) had significantly higher [SID] (52.9 ± 0.8 mEq/l) and PCO(2) (52.5 ± 0.7 mmHg) and relatively low [A(tot)] (14.9 ± 0.7 mEq/l) compared to average TCO(2) (32.1.0-34.9 mmol/l) horses (n = 75). In horses with the lowest TCO(2) (n = 11) the greatest contributor was elevated [A(tot) ] (21.0 ± 0.7 mEq/l) and unmeasured (acetate, citrate, proprionate, butyrate) weak acids (7.0 ± 0.2 mEq/l) while [SID] (49.6 ± 0.8 mEq/l) and PCO(2) (47.8 ± 1.0 mmHg) were similar to average TCO(2) horses. Thirty-two horses had a TCO(2) ranging from 35.0-36.9 mmol/l). CONCLUSIONS: There is a wide range of acid-base state and factors contributing to acid-base state amongst Standardbred race horses in Ontario. Dietary, environmental and handling practices and health concerns, that elevate plasma [SID], lower [A(tot)] and lower the concentration of unmeasured weak acids are the primary contributors to alkalosis and elevated TCO(2).

Nutritional aspects of post exercise skeletal muscle glycogen synthesis in horses: a comparative review.

Carbohydrate (CHO) stored in the form of skeletal muscle glycogen is the main energy source for glycolytic and oxidative ATP production during vigorous exercise in mammals. In man, horse and dog both short-term high intensity and prolonged submaximal exercise deplete muscle glycogen. In horses, however, muscle glycogen synthesis is 2-3-fold slower than in man and rat, even when a diet high in soluble CHO is fed. There appear to be significant differences in CHO and glycogen metabolism between horses and other mammals, and it is becoming increasingly clear that many conclusions drawn from human exercise physiology do not apply to horses. This review aims to provide a comprehensive, comparative summary of the research on muscle glycogen synthesis in horse, man and rodent. Species differences in CHO uptake and utilisation are examined and the issues with feeding high soluble CHO diets to horses are discussed. Alternative feeding strategies, including protein and long and short chain fatty acid supplementation and the importance of rehydration, are explored.