Hyperinsulinemic-euglycemic Clamp Procedure Research Articles

Effect of Weight Loss on Skeletal Muscle Bioactive Lipids in People With Obesity and Type 2 Diabetes.

Muscle sn-1,2-diacylglycerol (DAG) and C18:0 ceramide accumulation in sarcolemmal and mitochondrial compartments have been proposed to regulate muscle insulin sensitivity. Here, we evaluated whether weight loss-induced improvements in insulin sensitivity were associated with changes in muscle sn-1,2-DAG and ceramide content in people with obesity and type 2 diabetes. We measured skeletal muscle insulin sensitivity, assessed by using the hyperinsulinemic-euglycemic clamp procedure in conjunction with stable isotopically labeled glucose tracer infusion, and skeletal muscle sn-1,2-DAG and ceramide contents by using liquid chromatography-tandem mass spectrometry after subcellular fractionation and DAG isomer separation in 14 adults with obesity and type 2 diabetes before and after marked (18.6 ± 2.1%) weight loss. Whole-body insulin sensitivity doubled after weight loss. Sarcolemmal sn-1,2-DAG and C18:0 ceramide contents after weight loss were not different than values before weight loss. In contrast, mitochondrial/ER C18:0 ceramide content decreased by ∼20% after weight loss (from 2.16 ± 0.08 to 1.71 ± 0.13 nmol/g, P<0.005). These results suggest a decrease in muscle mitochondrial/ER C18:0 ceramide content could contribute to the beneficial effect of weight loss on skeletal muscle insulin sensitivity.

A single-center prospective analysis of the impact of glucose metabolism on pancreatic fistula onset after pancreaticoduodenectomy for periampullary tumors

BackgroundGlucose impairment notably affects the postoperative course of gastrointestinal surgeries. However, evidence on its impact on clinically relevant pancreatic fistulas(CR-POPFs) after pancreaticoduodenectomy(PD) is lacking. This study evaluates if and how preoperative glucose metabolism affects the development of CR-POPF after PD. MethodsOne hundred and ten consecutive PDs were included. Patients underwent preoperative metabolic profiling using the Oral Glucose Tolerance Test(OGTT) and the hyperinsulinemic euglycemic clamp procedure. Accordingly, patients were categorized as normal glucose tolerant (NGT), impaired glucose tolerant (IGT), diabetic (DM), and longstanding-DM. Receiver operating characteristics(ROC) analyses were performed to determine the values of metabolic features in prediction of CR-POPF. ResultsThe CR-POPF rate was 36.3 ​%(40 patients). NGT patients had a higher CR-POPF rate (51.7 ​%) compared to IGT(45.2 ​%), DM (15.8 ​%), and longstanding-DM (25.8 ​%) (p ​= ​0.03). CR-POPF patients had lower median fasting glucose levels (p ​= ​0.01) and higher c-peptide values at all OGTT time points (p ​< ​0.05). Fasting glucose and c-peptide levels had high diagnostic accuracy for CR-POPF (AUC>0.8) and were independent risk factors for CR-POPF (OR: 24.7[95%CI: 3.7–165.3] for fasting glucose; OR: 19.9[95%CI: 3.2–125.3] for c-peptide). ConclusionNormoglycemia and normal beta cell function may be risk factors for CR-POPF after PD. Fasting glucose and c-peptide levels effectively predicted CR-POPF development following PD. ClinicalTrials gov IdentifierNCT02175459

Adipocyte-specific inactivation of NAMPT, a key NAD+ biosynthetic enzyme, causes a metabolically unhealthy lean phenotype in female mice during aging.

Nicotinamide adenine dinucleotide (NAD+) is a universal coenzyme regulating cellular energy metabolism in many cell types. Recent studies have demonstrated the close relationships between defective NAD+ metabolism and aging and age-associated metabolic diseases. The major purpose of the present study was to test the hypothesis that NAD+ biosynthesis, mediated by a rate-limiting NAD+ biosynthetic enzyme, nicotinamide phosphoribosyltransferase (NAMPT), is essential for maintaining normal adipose tissue function and whole body metabolic health during the aging process. To this end, we provided in-depth and comprehensive metabolic assessments for female adipocyte-specific Nampt knockout (ANKO) mice during aging. We first evaluated body fat mass in young (≤4-mo-old), middle aged (10-14-mo-old), and old (≥18-mo-old) mice. Intriguingly, adipocyte-specific Nampt deletion protected against age-induced obesity without changing energy balance. However, data obtained from the hyperinsulinemic-euglycemic clamp procedure (HECP) demonstrated that, despite the lean phenotype, old ANKO mice had severe insulin resistance in skeletal muscle, heart, and white adipose tissue (WAT). Old ANKO mice also exhibited hyperinsulinemia and hypoadiponectinemia. Mechanistically, loss of Nampt caused marked decreases in WAT gene expression of lipogenic targets of peroxisome proliferator-activated receptor gamma (PPAR-γ) in an age-dependent manner. In addition, administration of a PPAR-γ agonist rosiglitazone restored fat mass and improved metabolic abnormalities in old ANKO mice. In conclusion, these findings highlight the importance of the NAMPT-NAD+-PPAR-γ axis in maintaining functional integrity and quantity of adipose tissue, and whole body metabolic function in female mice during aging.NEW & NOTEWORTHY Defective NAD+ metabolism is associated with aging and age-associated metabolic diseases. In the present study, we provided in-depth metabolic assessments in female mice with adipocyte-specific inactivation of a key NAD+ biosynthetic enzyme NAMPT and revealed an unexpected role of adipose tissue NAMPT-NAD+-PPAR-γ axis in maintaining functional integrity and quantity of adipose tissue and whole body metabolic health during the aging process.

NADPH oxidase-derived reactive oxygen species regulates adipose tissue lipolysis in humans

There is mounting evidence suggesting that reactive oxygen species (ROS) generated by NADPH oxidase (NOX) regulate physiological processes, as NOX has been shown to promote increases in lipolysis in cultured adipocytes. PURPOSE: To determine the mechanisms by which NOX stimulates adipose tissue lipolysis. METHODS: Young healthy females (n = 17; mean ± SD; age: 21.7 ± 5 years; body mass index: 22.2 ± 3.8 kg/m2) participated in this study. Participants underwent microdialysis and hyperinsulinemic-euglycemic clamp (clamp) procedures. Three microdialysis probes were inserted into subcutaneous abdominal adipose tissue to monitor local adipose tissue ROS (H2O2) production and lipolysis (as indicated by dialysate glycerol). To determine whether NOX stimulates lipolysis via distinct signaling pathways, microdialysis probes were perfused with either isoproterenol (Iso; β-adrenergic agonist to stimulate canonical lipolysis signaling) or atrial natriuretic peptide (ANP to stimulate non-canonical lipolysis signaling), then perfused with a local NOX inhibitor (apocynin). A control probe was perfused with apocynin but without any agonists of lipolysis. The microdialysis procedures were then repeated under hyperinsulinemic conditions during the same study visit. RESULTS: In the fasted state, dialysate H2O2 concentrations were lower in the control and Iso probes upon apocynin perfusion (control probe without apocynin: 1.06 ± 0.5 μM, with apocynin: 0.62 ± 0.2 μM; p = 0.005; Iso probe without apocynin: 1.01 ± 0.4 μM, with apocynin: 0.66 ± 0.2 μM; p = 0.003). Apocynin perfusion did not alter dialysate H2O2 concentrations in the ANP probe under the fasted state (p = 0.11). Dialysate glycerol concentrations did not change upon apocynin perfusion under any of the microdialysis probe conditions in the fasted state (p &gt; 0.05). During the clamp, apocynin perfusion resulted in lower dialysate H2O2 concentrations in the three conditions (control probe without apocynin: 1.22 ± 0.6 μM, with apocynin: 0.75 ± 0.3 μM; p = 0.03; Iso probe without apocynin: 1.55 ± 0.8 μM, with apocynin: 0.79 ± 0.3 μM; p = 0.01; ANP probe without apocynin: 1.55 ± 0.6 μM, with apocynin: 0.92 ± 0.3 μM; p = 0.03). Dialysate glycerol concentrations were lower in the Iso probe upon apocynin perfusion during the clamp (Iso probe without apocynin: 124.7 ± 38 μM, with apocynin: 90.9 ± 25 μM; p = 0.003). CONCLUSIONS: This study presents adipose tissue NOX as a source of in vivo ROS production in human participants. In addition, we reveal a novel mechanism of NOX in adipose tissue function whereby NOX exerts increases in β-adrenergic-mediated lipolysis during hyperinsulinemia. Our findings highlight the roles of ROS as signaling mediators and suggest that an imbalance between NOX activity and insulin action on adipose tissue could negatively affect systemic fatty acid metabolism. Porter Pre-doctoral Fellowship from the American Physiological Society and an NIH National Research Service Award (F31HL154642). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Clinical Pharmacology of GP40321 (Insulin Glulisine Biosimilar): Pharmacokinetic and Pharmacodynamic Comparability in a Hyperinsulinemic-Euglycemic Clamp Procedure.

The aim of the study was to compare the pharmacokinetics (PK) and pharmacodynamics (PD) of T-glu (GP40321, test drug), and reference insulin glulisine in a hyperinsulinemic-euglycemic clamp procedure. During this study, 34 healthy male volunteers underwent the hyperinsulinemic-euglycemic clamp procedure following subcutaneous 0.3U/kg injection of T-glu or reference insulin glulisine in a randomized, double-blind, crossover study. Plasma glucose levels were monitored every 5minutes for 8hours. Glucose infusion rate adjustment was based on the blood glucose measurements. Evaluation of PD was performed using the glucose infusion rate values, while PK was calculated using insulin concentrations measured via enzyme-linked immunosorbent assay. The study results showed that the 90% CI for the geometric mean ratios of primary PK and PD of T-glu and reference insulin glulisine were within 80%-125% comparability limits, and that the safety profiles were comparable. PK, PD, and safety similarity of T-glu and reference insulin glulisine was demonstrated.

Understanding the predictive accuracy of the InsuTAG index over other surrogate indices in normoglycaemic, non-obese males from Southern India

We aimed to evaluate the predictive accuracy of InsuTAG index against M value of the hyperinsulinaemic-Euglycaemic clamp (HEC) procedure and fasting surrogate indices of insulin sensitivity/resistance in young, normoglycaemic, Asian Indian males. HEC studies were done in young (mean age 19.7 ± 1 years), non-obese (mean BMI 19.2 ± 2.6 kg/m2), normoglycemic Asian Indian males (n = 110) and the M value was calculated. Surrogate indices namely InsuTAG index, HOMA-IR, FG-IR, QUICKI and McAuley index were calculated. Pearson’s correlation and ROC-AUC at 95% CI were applied. Significant negative correlation was observed for InsuTAG index with the M value (r − 0.23, p = 0.01), McAuley index (r − 0.65, p < 0.01), QUICKI (r − 0.34, p < 0.01) and FGIR (r − 0.35, p < 0.01). Significant positive correlations of InsuTAG index were observed for BMI and waist circumference. The ROC-AUC was higher for InsuTAG index (0.75) than FGIR (0.30), QUICKI (0.31), and McAuley index (0.20). The InsuTAG cut-off value ≥ 19.13 showed 66.7% sensitivity and 69.2% specificity in this study group.

Heterogeneity in the effect of marked weight loss on metabolic function in women with obesity.

BACKGROUNDThere is considerable heterogeneity in the effect of weight loss on metabolic function in people with obesity.METHODSWe evaluated muscle and liver insulin sensitivity, body composition, and circulating factors associated with insulin action before and after approximately 20% weight loss in women identified as "Responders" (n = 11) or "Non-responders" (n = 11), defined as the top (>75% increase) and bottom (<5% increase) quartiles of the weight loss-induced increase in glucose disposal rate (GDR) during a hyperinsulinemic-euglycemic clamp procedure, among 43 women with obesity (BMI: 44.1 ± 7.9 kg/m2).RESULTSAt baseline, GDR, which provides an index of muscle insulin sensitivity, and the hepatic insulin sensitivity index were more than 50% lower in Responders than Non-responders, but both increased much more after weight loss in Responders than Non-responders, which eliminated the differences between groups. Weight loss also caused greater decreases in intrahepatic triglyceride content and plasma adiponectin and PAI-1 concentrations in Responders than Non-responders and greater insulin-mediated suppression of plasma free fatty acids, branched-chain amino acids, and C3/C5 acylcarnitines in Non-responders than Responders, so that differences between groups at baseline were no longer present after weight loss. The effect of weight loss on total body fat mass, intra-abdominal adipose tissue volume, adipocyte size, and circulating inflammatory markers were not different between groups.CONCLUSIONThe results from our study demonstrate that the heterogeneity in the effects of marked weight loss on muscle and hepatic insulin sensitivity in people with obesity is determined by baseline insulin action, and reaches a ceiling when "normal" insulin action is achieved.TRIAL REGISTRATIONNCT00981500, NCT01299519, NCT02207777.FUNDINGNIH grants P30 DK056341, P30 DK020579, P30 DK052574, UL1 TR002345, and T32 HL13035, the American Diabetes Association (1-18-ICTS-119), the Longer Life Foundation (2019-011), and the Atkins Philanthropic Trust.

227-LB: Elevated Plasma Lactate and the Risk for Metabolic Syndrome—Is Skeletal Muscle a Producer or Consumer?

Background: The incidence of metabolic diseases is rising and there is a dire need for early detection to implement prevention. Fasting plasma lactate concentrations have been shown to be elevated in individuals with obesity and predictive of type 2 diabetes. An elevated fasting lactate concentration could possibly be the product of a reduction in muscle oxidative capacity and increased muscle lactate output. However, it is unknown how early this is evident, as are the mechanisms involved. Aim: The aim of this study was to determine if fasting lactate concentrations were linked with a reduction in skeletal muscle oxidative capacity in a cohort without metabolic disease. Methods: Young (age 28 ± 7; N=21) overweight (BMI (27.1 ± 0.4 kg/m2) men (n=12) and women (n=9) were recruited for this study. Blood biomarkers were measured in the fasted state and body composition was determined by DXA. Muscle oxidative capacity was measured in skeletal muscle homogenates (vastus lateralis biopsy) using 14C-labeled labeled palmitate and pyruvate. Muscle lactate output was determined by microdialysis during a hyperinsulinemic euglycemic clamp procedure. Results: Elevated lactate was associated with elevated visceral adiposity (R2=.40, p=.003) , elevated triglycerides (R2=.56, p=.0001) , as well as elevations in systolic blood pressure (R2=.39, p=.003) . Subjects with higher lactate had lower ratio of fatty acid oxidation to pyruvate oxidation (ratio R2=.49, p=.002) . Muscle lactate uptake in the basal state of the clamp positively associated with plasma lactate (R2=.92, p=&amp;lt;.0001) ; however, in the insulin phase net lactate output was near zero. Conclusion: Individuals who are overweight but otherwise healthy have elevated plasma lactate that associates with parameters of the metabolic syndrome. Lactate was associated with a shift from lipid to carbohydrate oxidation. Muscle is a consumer of lactate in the fasted state, whereas adipose tissue may be the source of production postprandially. Disclosure N. T. Broskey: None. E. J. Demaria: None. W. J. Pories: None. J. A. Houmard: None. L. Dohm: None. T. E. Jones: None. C. J. Tanner: None. D. Zheng: None. F. Jevtovic: None. P. Krassovskaia: None. L. E. Matarese: None. R. N. Cortright: None. R. Hickner: None. Funding NIH (R01DK120296)

561-P: The Effect of Time-Restricted Eating on Glycemic Control and Insulin Sensitivity in Women with Prediabetes: A Pilot Study

Time-restricted eating (TRE) is a dietary pattern in which daily food intake is restricted to a 6-h period without any caloric intake for the rest of the day. TRE improves glucose homeostasis in animal models even independent of weight loss and calorie intake. However, the weight-loss independent effect of TRE on glucose homeostasis in people remains debatable. We conducted a pilot study to investigate whether TRE without weight loss improves glycemic control and insulin sensitivity in women with prediabetes. Participants were randomized to the TRE (9-h daily eating period) or Control group (15-h daily eating period) for 12 weeks. We studied five participants (TRE n = 3 and Control n = 2) with overweight/obesity, prediabetes, and habitual daily eating period greater than 14 hours ([means and SD] age 50 ± 16 years, BMI 31 ± 5 kg/m2) . The daily eating period decreased in the TRE group (13.8 ± 0.5 vs. 8.5 ± 0.5 h) , but not in the Control group (13.9 ± 0.9 vs. 14.1 ± 1.1 h) . Participants’ weight, diet composition, and number of eating occasions remained unchanged during the study period for both groups. Glycemic control in free-living conditions assessed by using continuous glucose monitoring tended to improve in the TRE group (24-h area under the curve for interstitial glucose concentration: 2572 ± 286 vs. 2161 ± 216 mg/dl/24 h) , but not in the Control group (23± 59 vs. 2374 ± 2mg/dl/24 h) . Skeletal muscle insulin sensitivity assessed by using a hyperinsulinemic-euglycemic clamp procedure tended to increase in the TRE group (median and IQR: 27 (21, 37) vs. 34 (27, 41) μmol/kg/min) , but not in the Control group (41 ± 9 vs. 36 ± μmol/kg/min) . These preliminary results suggest that TRE may lead to weight-loss-independent improvements in glycemic control and insulin sensitivity in women with prediabetes. Further ongoing research will establish the effect of TRE without weight loss on glucose homeostasis and insulin sensitivity in people at high risk for type 2 diabetes mellitus. Disclosure M.Chondronikola: None. S.Zhu: None. P.Surampudi: n/a. B.Roshanravan: None. C.Sanchez: None. Funding NIH-UL1-TR001860, Clinical and Translational Science Center-Highly Innovative Pilot Award and USDA NationalInstitute of Food and Agriculture, Hatch project numberCA-D-NTR-2618-H

Increased plasma fatty acid clearance, not fatty acid concentration, is associated with muscle insulin resistance in people with obesity

BackgroundAlthough it is well-accepted that increased plasma free fatty acid (FFA) concentration causes lipid overload and muscle insulin resistance in people with obesity, plasma FFA concentration poorly predicts insulin-resistant glucose metabolism. It has been proposed that hyperinsulinemia in people with obesity sufficiently inhibits adipose tissue triglyceride lipolysis to prevent FFA-induced insulin resistance. However, we hypothesized enhanced FFA clearance in people with obesity, compared with lean people, prevents a marked increase in plasma FFA even when FFA appearance is high. MethodsWe assessed FFA kinetics during basal conditions and during a hyperinsulinemic-euglycemic clamp procedure in 14 lean people and 46 people with obesity by using [13C]palmitate tracer infusion. Insulin-stimulated muscle glucose uptake rate was evaluated by dynamic PET-imaging of skeletal muscles after [18F]fluorodeoxyglucose injection. ResultsPlasma FFA clearance was accelerated in participants with obesity and correlated negatively with muscle insulin sensitivity without a difference between lean and obese participants. Furthermore, insulin infusion increased FFA clearance and the increase was greater in obese than lean participants. ConclusionsOur findings suggest plasma FFA extraction efficiency, not just plasma FFA concentration, is an important determinant of the cellular fatty acid load and the stimulatory effect of insulin on FFA clearance counteracts some of its antilipolytic effect.

Dynamic assessment of insulin secretion and insulin resistance in Asians with prediabetes

Aims/hypothesisPrediabetes and type 2 diabetes are highly prevalent in Asia. Understanding the pathophysiology of abnormal glucose homeostasis in Asians will have important implications for reducing disease burden, but there have been conflicting reports on the relative contributions of insulin secretion and action in disease progression. In this study, we aimed to assess the contribution of β-cell dysfunction and insulin resistance in the Asian prediabetes phenotype. MethodsWe recruited 1679 Asians with prediabetes (n = 659) or normoglycemia (n = 1020) from a multi-ethnic population in Singapore. Participants underwent an oral glucose tolerance test, an intravenous glucose challenge, and a hyperinsulinemic-euglycemic clamp procedure to determine glucose tolerance, β-cell responsivity, insulin secretion, insulin clearance and insulin sensitivity. ResultsParticipants with prediabetes had significantly higher glucose concentrations in the fasting state and after glucose ingestion than did normoglycemic participants. Insulin sensitivity (M/I ratio) was ~15% lower, acute insulin response (AIR) to intravenous glucose and β-cell responsivity to oral glucose were ~35% lower, but total insulin secretion rate in the fasting state and after glucose ingestion was ~10% greater in prediabetic than in normoglycemic participants. The decrease in β-cell function with worsening glucose homeostasis in Asians with prediabetes was associated with progressively greater defects in AIR rather than M/I. However, analysis using static surrogate measures (HOMA indices) of insulin resistance and β-cell function revealed a different pattern. ConclusionsLower AIR to intravenous glucose and β-cell responsivity to oral glucose, on a background of mild insulin resistance, are the major contributors to the dysregulation of glucose homeostasis in Asians with prediabetes.

Lipid accumulation product (LAP) as a potential index to predict risk of insulin resistance in young, non-obese Asian Indian males from Southern India: observations from hyperinsulinemic-euglycemic clamp studies

IntroductionWe aimed to compare the predictive accuracy of surrogate indices namely the lipid accumulation product (LAP) index, homeostatic model of assessment of insulin resistance (HOMA-IR), fasting glucose-insulin ratio (FG-IR) and...

Differential insulin sensitivity of NMR-based metabolomic measures in a two-step hyperinsulinemic euglycemic clamp study

BackgroundInsulin is the key regulator of glucose metabolism, but it is difficult to dissect direct insulin from glucose-induced effects. We aimed to investigate the effects of hyperinsulemia on metabolomic measures under euglycemic conditions in nondiabetic participants.MethodsWe assessed concentrations of 151 metabolomic measures throughout a two-step hyperinsulinemic euglycemic clamp procedure. We included 24 participants (50% women, mean age = 62 [s.d. = 4.2] years) and metabolomic measures were assessed under baseline, low-dose (10 mU/m2/min) and high-dose (40 mU/m2/min) insulin conditions. The effects of low- and high-dose insulin infusion on metabolomic measures were analyzed using linear mixed-effect models for repeated measures.ResultsAfter low-dose insulin infusion, 90 metabolomic measures changed in concentration (p < 1.34e−4), among which glycerol (beta [Confidence Interval] = − 1.41 [− 1.54, − 1.27] s.d., p = 1.28e−95) and three-hydroxybutyrate (− 1.22 [− 1.36, − 1.07] s.d., p = 1.44e−61) showed largest effect sizes. After high-dose insulin infusion, 121 metabolomic measures changed in concentration, among which branched-chain amino acids showed the largest additional decrease compared with low-dose insulin infusion (e.g., Leucine, − 1.78 [− 1.88, − 1.69] s.d., P = 2.7e−295). More specifically, after low- and high-dose insulin infusion, the distribution of the lipoproteins shifted towards more LDL-sized particles with decreased mean diameters.ConclusionMetabolomic measures are differentially insulin sensitive and may thus be differentially affected by the development of insulin resistance. Moreover, our data suggests insulin directly affects metabolomic measures previously associated with increased cardiovascular disease risk.

Metabolically-Unhealthy Obesity Is Associated With Increased Adipose Tissue Inflammatory Gene Expression and 24-Hour Plasma Concentrations of PAI-1, but Not Other Inflammatory Cytokines

Insulin resistant glucose metabolism is the most common metabolic complication associated with obesity; however, a subset of people with obesity have normal insulin sensitivity and are considered to be metabolically healthy. In rodent models of obesity, adipose tissue (AT) inflammation contributes to whole-body insulin resistance mediated, at least in part, by production of proinflammatory cytokines that are secreted into the systemic circulation. We therefore hypothesized that AT markers of inflammation and plasma concentrations of inflammatory cytokines would be greater in people with metabolically-unhealthy obesity (MUO) and insulin-resistant glucose metabolism than in insulin-sensitive people with metabolically-healthy obesity (MHO). We measured AT expression of genes that encode for proinflammatory proteins by using RNA sequencing and plasma cytokine concentrations assessed serially over 24 hours by using multiplex assays in: i) 28 people with MHO (defined as normal glucose tolerance and normal insulin-stimulated glucose disposal assessed using the hyperinsulinemic-euglycemic clamp procedure [48 ± 2 µmol/kg fat-free mass/min]); and ii) 28 people with MUO (defined as prediabetes and impaired insulin-stimulated glucose disposal [28 ± 1 µmol/kg fat-free mass/min]). AT markers of inflammation (expression of SERPINE1, CCL3, CCL5, CD68, CD74, MRC1, and CXCL16) were greater in the MUO than in the MHO group (all P < 0.05). However, the 24-hour plasma concentration areas-under-the curve (AUC) for TNFα, MCP-1, IL-6, RANTES, IL-1β, IL-17, and IFN-γ were not different between MHO and MUO groups. In contrast, 24-hour plasma plasminogen activator inhibitor 1 (PAI-1) AUC was greater in the MUO (1,759 ± 169 ng/mL x h) group than in the MHO (716 ± 85 ng/mL x h) group (P < 0.001) and plasma PAI-1 was inversely correlated with whole-body insulin sensitivity (r= -0.57; P < 0.001). We conclude that, with the exception of PAI-1, AT inflammation does not contribute to whole-body insulin resistance by increasing systemic circulating inflammatory cytokine levels. However, increased AT production of PAI-1 is associated with whole-body insulin resistance in people with MUO.

Heterogeneity in insulin-stimulated glucose uptake among different muscle groups in healthy lean people and people with obesity

It has been proposed that muscle fibre type composition and perfusion are key determinants of insulin-stimulated muscle glucose uptake, and alterations in muscle fibre type composition and perfusion contribute to muscle, and consequently whole-body, insulin resistance in people with obesity. The goal of the study was to evaluate the relationships among muscle fibre type composition, perfusion and insulin-stimulated glucose uptake rates in healthy, lean people and people with obesity. We measured insulin-stimulated whole-body glucose disposal and glucose uptake and perfusion rates in five major muscle groups (erector spinae, obliques, rectus abdominis, hamstrings, quadriceps) in 15 healthy lean people and 37 people with obesity by using the hyperinsulinaemic-euglycaemic clamp procedure in conjunction with [2H]glucose tracer infusion (to assess whole-body glucose disposal) and positron emission tomography after injections of [15O]H2O (to assess muscle perfusion) and [18F]fluorodeoxyglucose (to assess muscle glucose uptake). A biopsy from the vastus lateralis was obtained to assess fibre type composition. We found: (1) a twofold difference in glucose uptake rates among muscles in both the lean and obese groups (rectus abdominis: 67 [51, 78] and 32 [21, 55] μmol kg-1min-1 in the lean and obese groups, respectively; erector spinae: 134 [103, 160] and 66 [24, 129] μmol kg-1min-1, respectively; median [IQR]) that was unrelated to perfusion or fibre type composition (assessed in the vastus only); (2) the impairment in insulin action in the obese compared with the lean group was not different among muscle groups; and (3) insulin-stimulated whole-body glucose disposal expressed per kg fat-free mass was linearly related with muscle glucose uptake rate (r2 = 0.65, p < 0.05). Obesity-associated insulin resistance is generalised across all major muscles, and is not caused by alterations in muscle fibre type composition or perfusion. In addition, insulin-stimulated whole-body glucose disposal relative to fat-free mass provides a reliable index of muscle glucose uptake rate.

The triglyceride/glucose ratio is a reliable index of fasting insulin resistance: Observations from hyperinsulinaemic-euglycaemic clamp studies in young, normoglycaemic males from southern India

Background & aimsNon-obese Asians have a high propensity to develop insulin resistance. Therefore, screening such individuals for insulin resistance using simple surrogate indices is important. In this study, we aimed to validate the triglyceride-glucose (Tg/glu) ratio against the M value of hyperinsulinaemic-euglycaemic clamp (HEC) procedure and other surrogate indices of insulin resistance in normoglycaemic Indian males from Southern India. MethodsA cohort of 105 normoglycaemic males (mean BMI: 19.2 ± 2.6 kg/m2) underwent HEC procedure. Surrogate indices of insulin resistance viz. the triglyceride-glucose (Tg/Glu) ratio, the triglyceride-glucose index, the McAuley’s index, the HOMA-IR, the QUICKI, the fasting glucose to insulin ratio (FG-IR), and the fasting C- peptide index were calculated and correlated with the M value. The cut-off value for the Tg/Glu ratio was obtained using the Receiver Operator Characteristics (ROC) with Area under curve (AUC) analysis at 95% confidence interval (CI). The P value < 0.05 was considered statistically significant. ResultsThe Tg/Glu ratio demonstrated significantly higher AUC (0.81), when compared to the Tg × glu index (0.63), 20/fasting C peptide × fasting plasma glucose index (0.55), HOMA-IR (0.47), QUICKI (0.26), FGIR (0.12) and McAuley’s index (0.18). For the Tg/Glu ratio, a cut-off value ≥ 1.19 had high sensitivity (80%) and specificity (79%) values (PPV: 16%; NPV: 98.8%) respectively. ConclusionThe Tg/Glu ratio can be used as a reliable surrogate index to screen for risk of insulin resistance in lean, normoglycaemic males from Southern India.

Obesity Is Associated With Increased Basal and Postprandial β-Cell Insulin Secretion Even in the Absence of Insulin Resistance.

We tested the hypothesis that obesity, independent of insulin resistance, is associated with increased insulin secretion. We compared insulin kinetics before and after glucose ingestion in lean healthy people and people with obesity who were matched on multiorgan insulin sensitivity (inhibition of adipose tissue lipolysis and glucose production and stimulation of muscle glucose uptake) as assessed by using a two-stage hyperinsulinemic-euglycemic pancreatic clamp procedure in conjunction with glucose and palmitate tracer infusions and positron emission tomography. We also evaluated the effect of diet-induced weight loss on insulin secretion in people with obesity who did not improve insulin sensitivity despite marked (∼20%) weight loss. Basal and postprandial insulin secretion rates were >50% greater in people with obesity than lean people even though insulin sensitivity was not different between groups. Weight loss in people with obesity decreased insulin secretion by 35% even though insulin sensitivity did not change. These results demonstrate that increased insulin secretion in people with obesity is associated with excess adiposity itself and is not simply a compensatory response to insulin resistance. These findings have important implications regarding the pathogenesis of diabetes because hyperinsulinemia causes insulin resistance and insulin hypersecretion is an independent risk factor for developing diabetes.

2010-P: Adipocyte Clusterin Is a Link between Cardiometabolism and Alzheimer’s Disease

Background: Obesity heightens the risk for both cardiometabolic complications and Alzheimer’s disease (AD). Although the reasons have yet to be fully elucidated, weight loss improves cognition. Given this link, factors overproduced by obese adipose tissue are ideal candidates for novel investigation into AD pathogenesis. We have recently shown that clusterin from the adipocyte is related to multiple factors of the metabolic syndrome (insulin resistance, dyslipidemia, NAFLD, and atherosclerosis). Clusterin is also involved in CNS lipid trafficking and has been proposed as an AD biomarker. Rationale: Our goals were to characterize the effect of high-fat diet (HFD) feeding on CNS clusterin and neuronal insulin signaling in an AD mouse model (5XFAD); and furthermore, determine the relationship of adipocyte clusterin with neurocognition and insulin sensitivity in obese humans at baseline and following bariatric surgery. Methods: We treated 5XFAD and WT mice with normal chow and HFD for 4 mo., measured CNS clusterin, and determined its effect on neuronal insulin signaling. We next measured subcutaneous adipocyte (SAd) clusterin expression, and performed neuropsychological evaluation and a hyperinsulinemic-euglycemic clamp procedure in obese human subjects (n=11) before and after sleeve gastrectomy. Results: In the 5XFAD mice, HFD increases brain clusterin levels compared to WT, and in primary cortical neurons clusterin attenuates insulin signaling in a dose-dependent manner. In humans, SAd clusterin is related to hepatic insulin resistance, higher fasting glucose, and reduced working memory/attention. After sleeve gastrectomy SAd clusterin expression is reduced, with a significant inverse association between clusterin and working memory/attention and hepatic insulin sensitivity. Conclusions: Our data suggests that SAd clusterin is a potential player in AD pathology, providing a critical link between excess adiposity, T2D, insulin resistance, and heightened AD risk. Disclosure D. Bradley: None. J.Z. Liu: None. A.M. Blaszczak: None. V.P. Wright: None. R. Tarawneh: None. Y. Liu: None. J. Ryu: None. S. Yoon: None. W. Hsueh: None. Funding American Diabetes Association (1-16-ICTS-049 to W.H.); National Institutes of Health (KL2TR001068, R01HL135622)

Heterogeneity in Insulin‐Stimulated Glucose Uptake Among Different Muscle Groups in People

Insulin‐stimulated glucose uptake in skeletal muscles is the major mechanism for postprandial glucose disposal. Insulin‐stimulated glucose uptake is greater in type I than in type II muscles in rats. The cellular capacity for glucose uptake and disposal (insulin receptor, GLUT4, hexokinase, and glycogen synthase expression) is also greater in isolated human type I compared with type II fibers. The goal of the present study was to test the hypothesis that insulin‐stimulated glucose uptake rate varies among different muscle groups and is greater in trunk (erector spinae, rectus abdominis, obliques) than leg (quadriceps, hamstrings) muscles and greater in the hamstrings than the quadriceps because trunk muscles generally have a greater percentage of type I fibers than leg muscles and the hamstrings have a greater percentage of type I fibers than quadriceps muscles. We determined abdominal and thigh muscle glucose uptake rates in 10 healthy lean women (age: 43±13 years, BMI: 23±2 kg/m2; mean±SD) by using a hyperinsulinemic‐euglycemic clamp procedure in conjunction with dynamic positron emission tomography imaging after bolus injection of 18F‐fluorodeoxyglucose. In addition, [6,6‐2H2]glucose was infused to determine whole‐body glucose disposal rate per kg fat‐free mass, which is commonly used as an index of muscle glucose uptake, and a muscle biopsy from the vastus lateralis was obtained to determine myofiber composition. Glucose uptake rate in the erector spinae (128±30 μmol/kg/min) was nearly twice as great as in the rectus abdominis (66±14 μmol/kg/min; P&lt;0.01) and approximately 50% greater than in the obliques (88±23 μmol/kg/min; P&lt;0.05). Quadriceps and hamstring glucose uptake rates were not different from each other (85±42 and 91±37 μmol/kg/min, respectively; P&gt;0.05) and not different from glucose uptake rates in the rectus abdominis and obliques but less than in the erector spinae (P&lt;0.05). No significant correlation between fiber type composition and muscle glucose uptake rate in the quadriceps was detected. Whole body insulin stimulated glucose disposal rate was 93±28 μmol/kg fat‐free mass/min. We conclude that: i) insulin‐stimulated muscle glucose uptake is heterogeneous among muscle groups, ii) it is unlikely that differences in fiber type composition among muscles are responsible for the heterogeneity in glucose uptake rates, and iii) whole‐body glucose disposal rate per kg fat‐free mass does not accurately reflect glucose uptake rates in individual muscles.Support or Funding InformationNIH DK115400, DK56341 (Nutrition Obesity Research Center), DK020579 (Diabetes Research Center), and UL1TR000448 (Clinical Translational Science Award), and ADA 1‐18‐ICTS‐119.

Insulin resistance drives hepatic de novo lipogenesis in nonalcoholic fatty liver disease

BACKGROUNDAn increase in intrahepatic triglyceride (IHTG) is the hallmark feature of nonalcoholic fatty liver disease (NAFLD) and is decreased by weight loss. Hepatic de novo lipogenesis (DNL) contributes to steatosis in individuals with NAFLD. The physiological factors that stimulate hepatic DNL and the effect of weight loss on hepatic DNL are not clear.METHODSHepatic DNL, 24-hour integrated plasma insulin and glucose concentrations, and both liver and whole-body insulin sensitivity were determined in individuals who were lean (n = 14), obese with normal IHTG content (n = 26), or obese with NAFLD (n = 27). Hepatic DNL was assessed using the deuterated water method corrected for the potential confounding contribution of adipose tissue DNL. Liver and whole-body insulin sensitivity was assessed using the hyperinsulinemic-euglycemic clamp procedure in conjunction with glucose tracer infusion. Six subjects in the obese-NAFLD group were also evaluated before and after a diet-induced weight loss of 10%.RESULTSThe contribution of hepatic DNL to IHTG-palmitate was 11%, 19%, and 38% in the lean, obese, and obese-NAFLD groups, respectively. Hepatic DNL was inversely correlated with hepatic and whole-body insulin sensitivity, but directly correlated with 24-hour plasma glucose and insulin concentrations. Weight loss decreased IHTG content, in conjunction with a decrease in hepatic DNL and 24-hour plasma glucose and insulin concentrations.CONCLUSIONSThese data suggest hepatic DNL is an important regulator of IHTG content and that increases in circulating glucose and insulin stimulate hepatic DNL in individuals with NAFLD. Weight loss decreased IHTG content, at least in part, by decreasing hepatic DNL.TRIAL REGISTRATIONClinicalTrials.gov NCT02706262.FUNDINGThis study was supported by NIH grants DK56341 (Nutrition Obesity Research Center), DK20579 (Diabetes Research Center), DK52574 (Digestive Disease Research Center), and RR024992 (Clinical and Translational Science Award), and by grants from the Academy of Nutrition and Dietetics Foundation, the College of Natural Resources of UCB, and the Pershing Square Foundation.