Localisation and composition of skeletal muscle diacylglycerol predicts insulin resistance in humans

We sought to evaluate if the cellular localisation and molecular species of diacylglycerol (DAG) were related to insulin sensitivity in human skeletal muscle. Healthy sedentary obese controls (Ob; n = 6; mean +/- SEM age 39.5 +/- 2.3 years; mean +/- SEM BMI 33.3 +/- 1.4 kg/m(2)), individuals with type 2 diabetes (T2D; n = 6; age 44 +/- 1.8 years; BMI 30.1 +/- 2.3 kg/m(2)), and lean endurance-trained athletes (Ath; n = 10; age 35.4 +/- 3.1 years; BMI 23.3 +/- 0.8 kg/m(2)) were studied. Insulin sensitivity was determined using an IVGTT. Muscle biopsy specimens were taken after an overnight fast, fractionated using ultracentrifugation, and DAG species measured using liquid chromatography/MS/MS. Total muscle DAG concentration was higher in the Ob (mean +/- SEM 13.3 +/- 1.0 pmol/mu g protein) and T2D (15.2 +/- 1.0 pmol/mu g protein) groups than the Ath group (10.0 +/- 0.78 pmol/mu g protein, p = 0.002). The majority (76-86%) DAG was localised in the membrane fraction for all groups, but was lowest in the Ath group (Ob, 86.2 +/- 0.98%; T2D, 84.2 +/- 1.2%; Ath, 75.9 +/- 2.7%; p = 0.008). There were no differences in cytoplasmic DAG species (p > 0.12). Membrane DAG species C18:0/C20:4, Di-C16:0 and Di-C18:0 were significantly more abundant in the T2D group. Cytosolic DAG species were negatively related to activation of protein kinase C (PKC)epsilon but not PKC theta, whereas membrane DAG species were positively related to activation of PKC epsilon, but not PKC theta. Only total membrane DAG (r = -0.624, p = 0.003) and Di-C18:0 (r = -0.595, p = 0.004) correlated with insulin sensitivity. Disaturated DAG species were significantly lower in the Ath group (p = 0.001), and significantly related to insulin sensitivity (r = -0.642, p = 0.002). These data indicate that both cellular localisation and composition of DAG influence the relationship to insulin sensitivity. Our results suggest that only saturated DAG in skeletal muscle membranes are related to insulin resistance in humans.