Insulin Dysregulation In Horses Research Articles

Pharmacokinetics and Alterations in Glucose and Insulin Levels After a Single Dose of Canagliflozin in Healthy Icelandic Horses.

Canagliflozin (CFZ) is a sodium-glucose cotransporter-2 inhibitor that has shown promising results as a drug for the treatment of insulin dysregulation in horses. Even though CFZ is used clinically, no pharmacokinetic data has previously been published. In this study, the pharmacokinetics of CFZ after administration of a single oral dose of 1.8 mg/kg in eight healthy Icelandic horses was examined. Additionally, the effect of treatment on glucose and insulin levels in response to a graded glucose infusion was investigated. Plasma samples for CFZ quantification were taken at 0, 0.33, 0.66, 1, 1.33, 1.66, 2, 2.33, 2.66, 3, 3.5, 4, 5, 6, 8, 12, 24, 32, and 48 h post administration. CFZ was quantified using UHPLC coupled to tandem quadrupole mass spectrometry (UHPLC-MS/MS). A non-compartmental analysis revealed key pharmacokinetic parameters, including a median Tmax of 7 h, a Cmax of 2350 ng/mL, and a t1/2Z of 28.5 h. CFZ treatment reduced glucose (AUCGLU, p = 0.001) and insulin (AUCINS, p = 0.04) response to a graded glucose infusion administered 5 h after treatment. This indicates a rapid onset of action following a single dose in healthy Icelandic horses. No obvious adverse effects related to the treatment were observed.

Sodium-glucose transport protein 2 inhibitor use in the management of insulin dysregulation in ponies and horses.

Laminitis is a common and painful condition of the equine foot and approximately 90% of cases are associated with insulin dysregulation (ID) that is a central feature of the common endocrine disorder equine metabolic syndrome (EMS) and occurs in a subset of animals with pituitary pars intermedia dysfunction. Additional features of EMS include obesity, altered circulating concentrations of adipokines (particularly adiponectin and leptin) and hypertriglyceridaemia. Obesity, ID, hypoadiponectinaemia, hyperleptinaemia and an altered plasma lipid profile are also features of human metabolic syndrome (HMS) alongside hyperglycaemia. Sodium-glucose cotransporter 2 inhibitors (SGLT2i) are a novel class of oral hypoglycaemic agents used in combination with lifestyle changes in the management of HMS. SGLT2 receptors are responsible for 90% of the renal glucose reabsorption that occurs in the proximal convoluted tubule. Thus, these drugs increase urinary glucose excretion by suppressing glucose reabsorption from the glomerular filtrate resulting in urinary calorie loss with consequent weight loss and improvements in ID, hyperglycemia, hypoadiponectinaemia and hyperleptinaemia. There are no licenced veterinary drugs available for treating ID and preventing insulin-associated laminitis in horses. Thus, the use of SGLT2i for the control of equine hyperinsulinaemia with the goal of improving recovery from associated active laminitis or preventing future laminitis has recently been advocated. There are a small number of published studies reporting the use of the SGLT2i canagliflozin, ertugliflozin and velagliflozin to aid the management of equine ID. However, the doses used are largely extrapolated from human studies with limited consideration of species-specific variations. In addition, there is limited evaluation of the fundamental differences between ID in horses and humans, particularly the fact that most horses with ID remain hyperinsulinaemic but normoglycaemic such that increased urinary loss of glucose may not explain the beneficial effects of these drugs. Further study of the potential deleterious effects of treatment-associated hypertriglyceridaemia is required together with the effect of SGLT2i therapy on circulating concentrations of adipokines in horses.

Focus on the epidemiology, pathophysiology, diagnosis, and management of insulin dysregulation in horses

Because there are various inter-related disorders of insulin metabolism that all may contribute to the final result of hyperinsulinaemia, the phrase "insulin dysregulation" has become the most acceptable term to use when describing equine metabolic syndrome (EMS). Insulin dysregulation is widespread in horses, and it appears to be more prevalent in physically sedentary horses. Laminitis and hyperinsulinaemia have been linked, as have other components of insulin dysregulation. Diagnosis of insulin dysregulation should be done in stages, beginning with basal testing, which includes baseline values of insulin and glucose, and concluding with dynamic tests, which detect tissue insulin resistance and hyperinsulinaemia. Dietary adjustments are the key to weight loss in the majority of EMS horses. To encourage weight loss, the horse should be given around 1.25% of its body weight every day. Exercise not only aids equid weight loss but also improves insulin sensitivity. The exercise should be tailored to the specific horse, taking into account its breed, fitness level, and owner's resources. When an obese horse is unable to exercise owing to laminitis or when obesity persists after extensive exercise and dietary management, levothyroxine sodium and metformin hydrochloride can be given to help accelerate weight loss and increase insulin sensitivity. Insulin dysregulation is a defining feature of equine metabolic syndrome and has garnered attention due to its direct link to laminitis. There is need for an in-depth understanding of ID in order to prevent the development of clinical laminitis. The current study discusses the epidemiology, pathophysiology, diagnosis, and management of insulin dysregulation in horses.

Evidence for dopamine production and distribution of dopamine D2 receptors in the equine gastrointestinal mucosa and pancreas.

Insulin dysregulation in horses is characterised by hyperinsulinaemia and/or tissue insulin resistance and is associated with increased risk of laminitis. There is growing evidence in other species that dopamine attenuates insulin release from the pancreas; however, this has yet to be examined in horses. The present study aimed to identify whether there are cells capable of producing or responding to dopamine within the equine gastrointestinal mucosa and pancreas. Tissue samples were collected from the stomach, small and large intestines, and pancreas of six mature horses following euthanasia. Samples of stomach contents and faeces were also collected. Immunohistochemistry was performed to identify tyrosine hydroxylase (TH), the rate-limiting enzyme for dopamine production, and dopamine D2 receptors in tissue sections. Additional immunostaining for glucagon, insulin and chromogranin A was performed to identify α cells, β cells and enteroendocrine cells, respectively. Gastric parietal cells expressed both TH and D2 receptors, indicating that they are capable of both producing and responding to dopamine. Dopamine was quantified in stomach contents and faeces by high-performance liquid chromatography with electrochemical detection, with similar concentrations found at both sites. Dopamine D2 receptors were expressed in duodenal epithelial cells but not more distally. A subset of enteroendocrine cells, located sporadically along the gastrointestinal tract, were found to be immunopositive for the D2 receptor. In pancreatic islets, TH was present in α cells, while D2 receptors were strongly expressed in β cells and variably expressed in α cells. These findings are consistent with studies of other species; however, dynamic studies are required to further elucidate the role of dopamine in the modulation of insulin and glucagon secretion in horses. This descriptive study provides preliminary evidence for a potential role of dopamine to act as a paracrine messenger in the gastrointestinal mucosa and endocrine pancreas of horses.

Effect of 5'-adenosine monophosphate-activated protein kinase agonists on insulin and glucose dynamics in experimentally induced insulin dysregulation in horses.

5'-adenosine monophosphate-activated protein kinase (AMPK) agonists, particularly resveratrol (RES), have not been extensively evaluated for their effect on insulin dysregulation (ID) in horses. Evaluate the effects of treatment with RES (10 mg/kg PO q12h), metformin (MET; 30 mg/kg PO q12h), and aspirin (ASP; 20 mg/kg PO q24h) on experimentally induced ID. Thirty-three healthy, adult, light-breed horses. Unblinded, placebo-controlled, experimental trial evaluating effects of AMPK agonists (RES, MET, and ASP) on experimentally induced ID. Horses were randomly assigned to a treatment group (RES, MET/ASP, RES/ASP, RES/MET/ASP, or placebo [CON]) after induction of ID with dexamethasone (0.08 mg/kg PO q24h for 7 days). Frequently sampled insulin-modified IV glucose tolerance tests (FSIGTT) and oral sugar tests (OST) were performed at baseline, 7 days after ID, and ID plus 7 days of treatment. Minimal model and OST variables were compared between (1-way ANOVA) and within (1-way ANOVA for repeated measures) groups over time to determine effects of treatment on ID. Administration of dexamethasone for 14 days resulted in significantly altered insulin and glucose dynamics (SI, DI, basal [glucose], and [insulin]) and produced clinical signs of laminitis in 5 out of 33 (15%) of horses included in the study. Combination therapy with RES, MET, and ASP did not significantly improve insulin and glucose dynamics in horses with experimentally induced ID. Metabolic testing before glucocorticoid administration should be considered in horses with clinical signs of metabolic syndrome.

Oligosaccharide feed supplementation reduces plasma insulin in geldings with Equine Metabolic Syndrome

BackgroundThe prevalence of Equine Metabolic Syndrome and insulin dysregulation is increasing in the horse population. Insulin dysregulation is a risk factor for laminitis, which has significant welfare consequences for the horse. Current treatment strategies for Equine Metabolic Syndrome have variable effects on insulin sensitivity. Findings suggest that gut microbiota plays an important role in gastrointestinal barrier protection and metabolic diseases. Dietary supplementation with oligosaccharides has been shown to increase the production of short-chain fatty acids and induce changes in the gut microbiota, with an increase in insulin sensitivity.ObjectivesWe hypothesized that dietary oligosaccharide supplementation would ameliorate insulin dysregulation in horses with Equine Metabolic Syndrome.Study designFifteen horses were included in a cross-over study including two feed supplementations periods with oligosaccharide supplementation or calcium carbonate. Before and after each period plasma insulin, endotoxin, serum amyloid A, and blood glucose concentrations were measured during an oral sugar test and fecal samples were collected.ResultsOligosaccharide supplementation significantly reduced insulin in geldings (p = 0.02). Overall, none of the dominating bacterias changed significantly in relative Δ-abundance. In the gut microbiota of mares, the Akkermansia genus and Clostridiaceae family were significantly more abundant. Within-sample bacterial diversity of horses with insulin concentrations > 60 mIU/L was significantly greater when compared to horses < 60 mIU/L. Horses with insulin concentrations > 60 mIU/L and horses with previous laminitis had a significantly greater beta diversity.Main limitationsCut-off values for oligosaccharide feed supplementation/placebo dosing instead of dosing by g/kg body weight; owner compliance; single blood sample in oral sugar test; inter-horse variationConclusionsThis study demonstrated that oligosaccharide supplementation could improve insulin dysregulation in geldings with Equine Metabolic Syndrome suggesting that sex susceptibility might be a factor to consider. Additionally, a tendency of reduction of obesity-associated bacteria was observed. Furthermore, the diversity of the gut microbiota was significantly dependent on laminitis status and insulin concentrations. However, more studies on the effects of oligosaccharide supplementation on insulin dysregulation are needed to improve the dietary management of horses with Equine Metabolic Syndrome.

Palatability, glycemic, and insulinemic responses to various carbohydrate formulations: Alternatives for the diagnosis of insulin dysregulation in horses?

Oral glycemic challenge (GC) tests are recommended for diagnosis of insulin dysregulation (ID). Various protocols are used, but all have limitations in terms of palatability, ease of use, variable composition, geographic availability, or some combination of these. To evaluate newly developed formulations with defined carbohydrate composition for use as oral GCs. Thirty-four horses and ponies in various metabolic states. Our objectives were carried out in 2 separate cross-over experiments. First, the palatability and acceptance of various GCs (2 syrups, 1 granulate) offered for free intake were compared to glucose mixed in a chaff-based diet. Subsequently, syrups were administered by syringe and compared to an oral glucose test using naso-gastric tubing (tube OGT) to investigate the glycemic and insulinemic responses. Second, these variables were compared in the best performing GC-formulations (granulate further optimized to pelleted formulation and 1 syrup) and a tube OGT. All GCs were administered with equivalent amounts of 0.5g glycemic carbohydrates per kg body weight. Only the GC pellets were consumed completely by all horses (consumption time 5 ± 2min). When administered by syringe, the GC syrup also was well accepted. The insulin concentrations at 120 min correlated significantly between tube OGT and GC pellets (r=.717; P < .001) or GC syrup (r=.913; P < .001). The new GC syrup and GC pellets discriminate between healthy and ID horses. The GC pellets (DysChEq)™ and GC syrup can be used as palatable and well-accepted oral GC tests for assessment of ID in horses.

Associations of plasma sphingolipid profiles with insulin response during oral glucose testing in Icelandic horses.

BackgroundSphingolipids modulate insulin sensitivity in mammals. Increased synthesis of ceramides is linked to decreased insulin sensitivity of tissues. Conversely, activation of the insulin signaling pathway can downregulate ceramide synthesis. Elucidating the association between sphingolipid metabolism and insulin response during oral glucose testing may help explain the pathophysiology of insulin dysregulation in horses.HypothesesHorses with insulin dysregulation will have a plasma sphingolipid profile characterized by increased ceramide concentrations. The plasma sphingolipid profile will have decreased ceramide concentrations after acute activation of the insulin signaling pathway by oral glucose testing.AnimalsTwelve Icelandic horses.MethodsHorses were subjected to an oral glucose test (0.5 g/kg body weight glucose), with plasma insulin concentrations measured at 0, 30, 60, 120, 180, and 240 minutes postglucose administration. Plasma samples were collected at 0 and 120 minutes for sphingolipid profiling using a liquid chromatography‐mass spectrometry‐based metabolomics analysis. Eighty‐three species of sphingolipids were detected, including 3‐ketosphinganines, dihydroceramides, ceramides, dihydrosphingomyelins, sphingomyelins, galatosylceramides, glucosylceramides, lactosylceramides, and ceramide‐1‐phosphates.ResultsGlucose administration did not significantly alter plasma sphingolipid profiles. C22:0‐ceramide, C24:1‐ceramide, C23:0‐ceramide, C16:1‐sphingomyelin, C22:0‐dihydroceramide, and C24:0‐ceramide were positively correlated with the insulin response (area under the curve).Conclusion and Clinical ImportancePositive correlation between the insulin response and sphingolipid concentrations implies upregulated sphingolipid metabolism in insulin dysregulated horses. A high plasma ceramide concentration can indicate insulin dysregulation in horses.

Effects of advanced age and pituitary pars intermedia dysfunction on components of the acute phase reaction in horses

Age, neurodegenerative disorders, and dysfunction of insulin secretion may be correlated with increased systemic concentrations of acute phase markers. Thus, the study aimed to determine the effect of age, pituitary pars intermedia dysfunction (PPID), and insulin dysregulation (ID) associated with PPID, on markers of the acute phase reaction. Twenty-nine mix-breed horses of both sexes were classified into groups: (1) healthy adult controls, (2) healthy non-PPID geriatric horses, (3) PPID ID+ horses, and (4) PPID ID− horses. Whole blood proinflammatory cytokine gene expression and serum concentrations of pro- and anti-inflammatory cytokines and acute phase proteins were measured. The data were analyzed using the Mann–Whitney U-test, and correlations between groups of data were assessed using Spearman's correlation coefficient. The tests were statistically significant if P < 0.05. No differences in the whole blood cytokine gene expression, serum cytokine concentrations, or acute phase proteins were noted between the groups. In the PPID ID group, there was a strong correlation between the ACTH concentration after the administration of thyrotropin-releasing hormone and the expression of IL-8 (r = 0.941; P = 0.0321). In the PPID ID+ group, there was a strong correlation between basal insulin concentrations and serum amyloid A (SAA; r = 0.936; P = 0.0083) as well as between postprandial insulin concentrations and SAA (r = 0.965; P = 0.001). These data suggest that neurodegeneration in horses moderately affects circulating markers of inflammation and that ID in horses with PPID influences acute phase inflammatory markers.

Comparison of Two Diagnostic Methods to Detect Insulin Dysregulation in Horses Under Field Conditions

Straightforward testing procedures to enable the diagnosis of insulin dysregulation (ID) in horses that are suitable for use in daily veterinary practice are needed because of the risk that ID could result in laminitis. In our study (that included 90 horses), we compared the proportion of horses classified as ID-positive, ID-suspect, and ID–not diagnosed according to the basal insulin concentration (BIC) with the proportion of horses classified as ID-positive or ID-negative according to a practical and feasible version of an oral sugar test (OST). Furthermore, BIC, basal glucose concentration, and insulin and glucose concentration after OST were analyzed and compared. In the total study population, the OST detected significantly more ID-positive cases than the BIC, with cutoffs at equivalent specificities. Receiver operating characteristics analysis showed that at a lower cutoff, the sensitivity of the BIC could be increased, but at the cost of a significantly lower specificity. Taking this into account, we found diagnostic performance of the OST to be considerably better than the BIC and therefore considered it more recommendable for use as a screening test for ID in ambulatory practice. Furthermore, we investigated the relationship between body condition score and breed type with glucose and insulin concentration as determined after our version of the OST. For that purpose, the study group was subdivided into lean, moderate, and obese horses and “easy keeper breeds” versus “non-easy keeper breeds”. Results supported the general assumption that obese horses and “easy keeper breeds” are more prone to the development of ID.

Insulin dysregulation in horses with induced obesity

ABSTRACT: Insulin deregulation (ID) is a central player in the pathophysiology of equine metabolic syndrome (EMS), which is associated with generalized and/or regional obesity. The objective of this experiment was to characterize the alterations in the hormonal profile in horses exposed to a hypercaloric diet. A total of nine Mangalarga Marchador adult horses with initial body condition score (BCS) of 2.9±1/9 (mean±SD) were submitted to a high calorie grain-rich diet for 5 months. The data was collected before the start of the experiment and every 15 days until the end of the experiment and glucose and insulin concentrations were measured in the plasma. Proxies G:I, RISQI, HOMA-IR and MIRG were calculated. The low-dose oral glucose tolerance test (OGTT) was performed and the total area under the glucose (GTA) and insulin (ITA) curves at three different timepoints (before inducing obesity, after 90 days and after 150 days) was used. Analysis of variance of the results was performed considering the time effects and the means were compared with repeated measures by the Tukey’s test (P≤0.05). The ID was observed during the first 90 days of the experiment and was characterized as a decompensated ID, showing an increase of basal glucose and insulin plasma levels, changes in all proxies and a significant increase in GTA (P&lt;0.001) and ITA (P&lt;0.05). However, a clear compensation of the ID was evident after 150 days of experiment, which was supported by data from the insulin secretory response of β cells of the pancreas that showed an increase in insulin plasma levels, after fasting or exposure to gastric glucose, with a concomitant decrease in fasting glucose and fructosamine levels, and a decrease of GTA and marked increase of ITA (P&lt;0.0001) in the dynamic test. These findings confirm the occurrence of hyperinsulinemia associated with insulin deregulation in Mangalarga Marchador horses exposed to hypercaloric diets.

Evaluation of combined testing to simultaneously diagnose pituitary pars intermedia dysfunction and insulin dysregulation in horses.

BackgroundThe thyrotropin‐releasing hormone (TRH) stimulation test and the 2‐step insulin sensitivity test are commonly used methods to diagnose, respectively, pituitary pars intermedia dysfunction (PPID) and insulin dysregulation (ID).ObjectivesTo investigate the diagnostic value of combining the TRH stimulation test and the 2‐step insulin sensitivity test to diagnose PPID and ID simultaneously.AnimalsTwenty‐seven adult horses, 10 control horses without PPID or ID, 5 horses with PPID only, 5 horses with ID only, and 7 horses with PPID and ID.MethodsRandomized prospective study. Horses underwent a TRH stimulation test alone, a 2‐step insulin sensitivity test alone, and combined testing with simultaneous TRH and insulin injection in the same syringe. Data were compared by 2‐way repeated measures analysis of variance and 2 1‐sided tests to demonstrate equivalence. Bland‐Altman plots were generated to visualize agreement between combined and independent testing.ResultsThe effect of combined testing on plasma adrenocorticotropic hormone, blood glucose concentration, or percentage decrease in blood glucose concentration was not significantly different from the effect obtained with independent testing. One control horse appeared falsely positive for PPID, 2 PPID‐only horses appeared falsely positive for ID, and 1 PPID and ID horse appeared falsely negative for ID when tests were performed simultaneously. Bland‐Altman plots supported the agreement between combined and independent testing.Conclusions and Clinical ImportanceCombining the TRH stimulation test and the 2‐step insulin sensitivity test appears to be a useful diagnostic tool for equine practitioners in the field, allowing testing of a horse for both PPID and ID simultaneously.

Factors associated with survival, laminitis and insulin dysregulation in horses diagnosed with equine pituitary pars intermedia dysfunction.

Pituitary pars intermedia dysfunction (PPID) is a commonly described endocrine disorder in higher latitudes of the Northern hemisphere but the description of the disease at lower latitudes and in the Southern hemisphere is limited. Document the clinical features of PPID at different Australian latitudes and climates, and investigate factors associated with survival, laminitis and insulin dysregulation (ID). Retrospective study of 274 equids from eight institutions across Australia. A diagnosis of PPID was based on endogenous ACTH, overnight dexamethasone suppression test, thyrotropin-releasing hormone stimulation test or necropsy. Clinical and clinicopathologic characteristics of PPID and therapeutic responses were investigated. Laminitis was diagnosed by radiographic or histologic changes and ID was diagnosed based on endogenous insulin, an oral glucose test or a 2-step insulin-response test. Being a pony, having a higher body condition score and pergolide administration were associated with survival. The clinical presentation of PPID changed with latitude and climate, with anhidrosis and polyuria/polydipsia more commonly recognised at lower latitudes. Laminitis was diagnosed in 89.9% of cases and ID was present in 76.5% of cases in which they were investigated. Despite the sample size, the lack of uniform testing at all locations (primary or referral cases) and the incompleteness of data sets limited the power of the statistical analyses. PPID can present with variable signs at different latitudes and climates, and ID should be investigated in equids diagnosed with PPID. Adequate body condition and administration of pergolide are fundamental in PPID management.

Pharmacokinetics of Metformin in Combination With Sitagliptin in Adult Horses After Enteral Administration

Insulin dysregulation (ID) is a common metabolic disorder in horses. Recently, incretin hormone release has been suggested to be involved in ID in horses. In human medicine, metformin and sitagliptin are commonly used in combination for metabolic syndrome. This combination could be useful in treating ID in horses. However, no pharmacokinetics data have been reported in this species. The objective of the present study was to establish the plasma concentration–time profile and to derive pharmacokinetics data for a combination of metformin and sitagliptin in horses after enteral administration. Six healthy adult Purebred Spanish horses were used. A metformin (15 mg/kg) plus sitagliptin (1.5 mg/kg) preparation was administered by intragastric route (IG) as an enteral solution. Blood samples were collected from 0 to 48 hours after IG drug administration. Plasma concentrations of metformin and sitagliptin were measured using high performance liquid chromatography methods. The t½λz for metformin was 2.9 hours and for sitagliptin 21 hours. The Cmax was 442 ± 84 mg/L within 0.9 hours for metformin and 94 ± 14 mg/L within 1.3 hours for sitagliptin. No adverse effects were observed, and the combination of metformin and sitagliptin was well tolerated. Therefore, these results suggest that metformin plus sitagliptin might be a combination to consider in horses with ID. Additional studies are needed to establish the effectiveness and tolerance in equids affected by endocrine disorders.

Insulin dysregulation in horses with systemic inflammatory response syndrome.

BackgroundSystemic inflammation is a cause of insulin dysregulation in many species, but the insulin and glucose dynamics in adult horses diagnosed with systemic inflammatory response syndrome (SIRS) are poorly documented.Hypothesis/ObjectivesIn SIRS in horses, insulin and glucose dynamics will be altered and associated with survival.AnimalsAdult horses diagnosed with SIRS admitted to a referral hospital.MethodsProspective study enrolling horses diagnosed with SIRS in which serum insulin and glucose concentrations were measured. Horses were grouped by outcome (survival, hyperinsulinemia, and hyperglycemia) and compared with P < .05 considered significant.ResultsFifty‐eight horses were included in the study and 36 (62%) survived. At admission, 21 horses (36%) were hyperinsulinemic and 44 horses (88%) were hyperglycemic, with survivors having significantly higher serum insulin and a significantly lower serum glucose concentration. Horses diagnosed with hyperinsulinemia at any time during hospitalization were 4 times more likely to survive whereas horses that were hyperglycemic at any time during hospitalization were 5 times less likely to survive. Serum glucose concentration and presence of hyperglycemia both were associated with severity of disease. Insulin/glucose ratio, reflecting insulin secretion, was significantly higher in survivors whereas glucose/insulin ratio, reflecting peripheral tissue insulin resistance, was significantly lower in nonsurvivors. Only in survivors was there a significant correlation between serum insulin and glucose concentrations.Conclusions and Clinical ImportanceHyperinsulinemia and hyperglycemia are common features of SIRS in horses, but those presenting with relative hypoinsulinemia and corresponding hyperglycemia suggestive of endocrine pancreatic dysfunction have a worse prognosis.

Lower plasma trans-4-hydroxyproline and methionine sulfoxide levels are associated with insulin dysregulation in horses

BackgroundInsulin dysregulation in horses is a metabolic condition defined by high insulin concentrations in the blood and peripheral insulin resistance. This hyperinsulinemia is often associated with severe damage in the hooves, resulting in laminitis. However, we currently lack detailed information regarding the potential involvement of particular metabolic pathways in pathophysiological causes and consequences of equine insulin dysregulation. This study aimed to assess the dynamic metabolic responses given to an oral glucose test (OGT) in insulin-sensitive and insulin-dysregulated horses by a targeted metabolomics approach to identify novel metabolites associated with insulin dysregulation.ResultsOral glucose testing triggered alterations in serum insulin (26.28 ± 4.20 vs. 422.84 ± 88.86 μIU/mL, p < 0.001) and plasma glucose concentrations (5.00 ± 0.08 vs. 9.43 ± 0.44 mmol/L, p < 0.001) comparing basal and stimulated conditions after 180 min. Metabolome analyses indicated OGT-induced changes in short-chain acylcarnitines (6.00 ± 0.53 vs. 3.99 ± 0.23 μmol/L, p < 0.001), long-chain acylcarnitines (0.13 ± 0.004 vs. 0.11 ± 0.002 μmol/L, p < 0.001) and amino acids (2.18 ± 0.11 vs. 1.87 ± 0.08 μmol/L, p < 0.05). Kynurenine concentrations increased (2.88 ± 0.18 vs. 3.50 ± 0.19 μmol/L, p < 0.01), whereas spermidine concentrations decreased during OGT (0.09 ± 0.004 vs. 0.08 ± 0.002 μmol/L, p < 0.01), indicating proinflammatory conditions after oral glucose load. Insulin dysregulation was associated with lower concentrations of trans-4-hydroxyproline (4.41 ± 0.29 vs. 6.37 ± 0.71 μmol/L, p < 0.05) and methionine sulfoxide (0.40 ± 0.06 vs. 0.87 ± 0.13 μmol/L, p < 0.01; mean ± SEM in insulin-dysregulated vs. insulin-sensitive basal samples, respectively), two metabolites which are related to antioxidant defense mechanisms.ConclusionOral glucose application during OGT resulted in profound metabolic and proinflammatory changes in horses. Furthermore, insulin dysregulation was predicted in basal samples (without OGT) by pathways associated with trans-4-hydroxyproline and methionine sulfoxide, suggesting that oxidative stress and oxidant–antioxidant disequilibrium are contributing factors to insulin dysregulation. The present findings provide new hypotheses for future research to better understand the underlying pathophysiology of insulin dysregulation in horses.

Assessment of Owner and Veterinarian Awareness of Equine Insulin Dysregulation and Available Treatments in Southeastern United States

Insulin dysregulation (ID) refers collectively to horses afflicted with fasting hyperinsulinemia and insulin resistance. Although there is no current data indicating the percentage of equines afflicted with ID, it appears to be a common subject among equine enthusiasts. In order to gauge public awareness of ID in horses and its available treatments, 122 horse owners and 18 veterinarians, representing 25 states, participated in a self-administered online survey. Questions were postulated to ascertain participant's awareness of the symptoms, management techniques, current treatments and medications, and knowledge acquisition associated with ID. The study revealed more than half of participants were “moderately aware” (34%) to “very aware” (25%) of ID in horses; however, when identifying symptoms of ID, some reported symptoms of pituitary pars intermedia dysfunction as those of ID. Many owners recognized current management strategies for ID; however, actual implementation strategies did not reflect standard management methods. Treatments mentioned for ID include supplements (14% of owners), thyroid supplement Thyro-L (13% and 72%), and pergolide/Prascend (21% and 61%) of owners and veterinarians, respectively. Veterinarians recommended diet and exercise as their main management strategy. Both owners and veterinarians reported using published research and veterinarians as a main source of information. The findings of this survey suggest that although awareness and recognition of the condition is high, proper identification, treatment, and management of ID varies. Moreover, research addressing the efficacy of pergolide as a treatment for ID is lacking. Additional education and research is needed to address these shortcomings.

The diagnosis of equine insulin dysregulation.

Insulin dysregulation is the hallmark of equine metabolic syndrome and has received attention because of its direct association with laminitis. In the absence of an adequate treatment for laminitis, a focus on prophylaxis is needed, making early detection of individuals at risk of developing laminitis one of the main challenges in equine endocrinology. Recent studies have shown that insulin dysregulation goes beyond tissue insulin resistance and it is now demonstrated that the equine enteroinsular axis plays a major role in insulin secretion and equine hyperinsulinaemia. In this review, we discuss the different tests currently available to diagnose insulin dysregulation in horses: the ones investigating tissue insulin resistance and those investigating the enteroinsular axis, detailing their goals, practicalities and limitations. This review supports the contention that the diagnosis of equine insulin dysregulation should now be based on the investigation of both tissue insulin resistance and the equine enteroinsular axis. Regardless of the tests used many factors of variation, such as breed, diet, fasting state or season, have been identified and could potentially confound the results of a specific test. Therefore, careful interpretation of the results of a given test in each individual situation is required to optimise the detection of horses at risk of laminitis.

A modified oral sugar test for evaluation of insulin and glucose dynamics in horses.

BackgroundAn oral sugar test (OST) using Karo® Light Corn Syrup has been developed in the USA as a field test for the assessment of insulin dysregulation in horses but the syrup is not available in Scandinavian grocery stores. The aim of the study was to compare the results of a modified OST between horses with equine metabolic syndrome (EMS) and healthy horses using a Scandinavian commercially available glucose syrup (Dansukker glykossirap). In addition, the effect of breed and the repeatability of the test were evaluated. In the present study, clinically healthy horses (7 Shetland ponies, 8 Icelandic horses, 8 Standardbred horses) and 20 horses of various breeds with EMS underwent the modified OST test. The Icelandic horses and Shetland ponies underwent the OST twice. Insulin and glucose data from the OST were used to calculate peak insulin concentration (PeakINS), time to peak insulin concentration (T-peakINS), area under the curve for insulin (AUCINS) and glucose (AUCGLU) as well as whole body insulin sensitivity index (ISICOMP).ResultsCompared to the healthy group, the EMS group had 6–7 times higher geometric mean for PeakINS and AUCINS and 8 times lower geometric mean for ISICOMP. The EMS group had a delayed T-peakINS compared to the healthy group. There was no effect of breed in the group of healthy horses on PeakINS, T-peakINS, AUCINS, AUCGLU and ISICOMP. Coefficient of variation for repeated tests was 19.8, 19.0 and 17.6 % for PeakINS, AUCINS and ISICOMP respectively.ConclusionsThe results of the present study demonstrate that the modified OST appears to be a practical and useful diagnostic tool for assessment of insulin dysregulation in the horse. However, to make it possible to establish the most appropriate sampling interval and to evaluate the accuracy of the modified OST, further studies in horses with a variable degree of insulin resistance are needed, where results from the modified OST are compared with quantitative measurements for IS.

Comparison of three different methods for the quantification of equine insulin.

BackgroundExact analysis of equine insulin in blood samples is the key element for assessing insulin resistance or insulin dysregulation in horses. However, previous studies indicated marked differences in insulin concentrations obtained from sample analyses with different immunoassays. Most assays used in veterinary medicine are originally designed for use in human diagnostics and are based on antibodies directed against human insulin, although amino acid sequences between equine and human insulin differ. Species-specific assays are being used more frequently and seem to provide advantages compared to human-specific assays. The aim of this study was to compare three immunoassays, one porcine-specific insulin enzyme-linked immunosorbent assay (ELISA), advertised to be specific for equine insulin, one porcine-specific insulin radioimmunoassay (RIA) and one human-specific insulin chemiluminescence immunoassay (CLIA), all three widely used in veterinary laboratories for the analysis of equine insulin. Furthermore, we tested their clinical applicability in assessing insulin resistance and dysregulation by analysis of basal blood and blood samples obtained during a dynamic diagnostic stimulation test (OGT) with elevated insulin concentrations.ResultsInsulin values obtained from the ELISA, RIA and CLIA, investigated for analyses of basal blood samples differed significantly between all three assays. Analyses of samples obtained during dynamic diagnostic stimulation testing with consecutively higher insulin concentrations revealed significantly (p < 0.001) lower insulin concentrations supplied by the CLIA compared to the ELISA. However, values measured by ELISA were intermediate and not different to those measured by RIA. Calculated recovery upon dilution, as a marker for assay accuracy in diluted samples, was 98 ± 4 % for ELISA, 160 ± 41 % for RIA and 101 ± 11 % for CLIA.ConclusionsOur results indicate that insulin concentrations of one sample measured by different methods vary greatly and should be interpreted carefully. Consideration of the immunoassay method and reliable assay-specific reference ranges are of particular importance especially in clinical cases where small changes in insulin levels can cause false classification in terms of insulin sensitivity of horses and ponies.