Altered tricarboxylic acid cycle flux in primary myotubes from severely obese humans

• Published in: International Journal of Obesity 2019-04, Vol.43 (4), p.895-905
• Main Authors: Zou, Kai, Hinkley, J. Matthew, Park, Sanghee, Zheng, Donghai, Jones, Terry E., Pories, Walter J., Hornby, Pamela J., Lenhard, James, Dohm, G. Lynis, Houmard, Joseph A.
• Format: Article
• Language: English
• Subjects: 13; 13/106; 631/443/319/1642/393; 692/699/2743/393; Adult; Backup software; Carbohydrate Metabolism - physiology; Carbon 14; Cells, Cultured - physiology; Citric Acid Cycle - physiology; Complications and side effects; Diabetes mellitus; Diabetes mellitus (non-insulin dependent); Epidemiology; Female; Fluctuations; Flux; Glucose; Glucose metabolism; Glycogen; Glycogen - metabolism; Glycogen synthesis; Glycolysis; Glycolysis - physiology; Health aspects; Health Promotion and Disease Prevention; Humans; Insulin; Intermediates; Internal Medicine; Krebs cycle; Male; Medicine; Medicine & Public Health; Metabolic Diseases; Metabolomics; Muscle Fibers, Skeletal - metabolism; Muscle Fibers, Skeletal - pathology; Muscles; Musculoskeletal system; Myotubes; Obesity; Obesity, Morbid - metabolism; Obesity, Morbid - physiopathology; Oxidation; Oxidation-reduction reactions; Partitioning; Physiological aspects; Polysaccharides; Primary Cell Culture; Proteins; Public Health; Pyruvic acid; Skeletal muscle; Synthesis; Tricarboxylic acid cycle; Tricarboxylic Acids - metabolism; Type 2 diabetes
• ISSN: 0307-0565; 1476-5497
• DOI: 10.1038/s41366-018-0137-7

Background/objective The partitioning of glucose toward glycolytic end products rather than glucose oxidation and glycogen storage is evident in skeletal muscle with severe obesity and type 2 diabetes. The purpose of the present study was to determine the possible mechanism by which severe obesity alters insulin-mediated glucose partitioning in human skeletal muscle. Subjects/methods Primary human skeletal muscle cells (HSkMC) were isolated from lean (BMI = 23.6 ± 2.6 kg/m 2 , n  = 9) and severely obese (BMI = 48.8 ± 1.9 kg/m 2 , n  = 8) female subjects. Glucose oxidation, glycogen synthesis, non-oxidized glycolysis, pyruvate oxidation, and targeted TCA cycle metabolomics were examined in differentiated myotubes under basal and insulin-stimulated conditions. Results Myotubes derived from severely obese subjects exhibited attenuated response of glycogen synthesis (20.3%; 95% CI [4.7, 28.8]; P  = 0.017) and glucose oxidation (5.6%; 95% CI [0.3, 8.6]; P  = 0.046) with a concomitant greater increase (23.8%; 95% CI [5.7, 47.8]; P  = 0.004) in non-oxidized glycolytic end products with insulin stimulation in comparison to the lean group (34.2% [24.9, 45.1]; 13.1% [8.6, 16.4], and 2.9% [−4.1, 12.2], respectively). These obesity-related alterations in glucose partitioning appeared to be linked with reduced TCA cycle flux, as 2-[ 14 C]-pyruvate oxidation (358.4 pmol/mg protein/min [303.7, 432.9] vs. lean 439.2 pmol/mg protein/min [393.6, 463.1]; P  = 0.013) along with several TCA cycle intermediates, were suppressed in the skeletal muscle of severely obese individuals. Conclusions These data suggest that with severe obesity the partitioning of glucose toward anaerobic glycolysis in response to insulin is a resilient characteristic of human skeletal muscle. This altered glucose partitioning appeared to be due, at least in part, to a reduction in TCA cycle flux.