Protein Ingestion Induces Muscle Insulin Resistance Independent of Leucine-Mediated mTOR Activation

Gordon I Smith, Jun Yoshino, Kelly L Stromsdorfer, Seth J Klein, Faidon Magkos, Dominic N Reeds, Samuel Klein, Bettina Mittendorfer, Gordon I Smith, Jun Yoshino, Kelly L Stromsdorfer, Seth J Klein, Faidon Magkos, Dominic N Reeds, Samuel Klein, Bettina Mittendorfer

Abstract

Increased plasma branched-chain amino acid concentrations are associated with insulin resistance, and intravenous amino acid infusion blunts insulin-mediated glucose disposal. We tested the hypothesis that protein ingestion impairs insulin-mediated glucose disposal by leucine-mediated mTOR signaling, which can inhibit AKT. We measured glucose disposal and muscle p-mTOR(Ser2448), p-AKT(Ser473), and p-AKT(Thr308) in 22 women during a hyperinsulinemic-euglycemic clamp procedure with and without concomitant ingestion of whey protein (0.6 g/kg fat-free mass; n = 11) or leucine that matched the amount given with whey protein (n = 11). Both whey protein and leucine ingestion raised plasma leucine concentration by approximately twofold and muscle p-mTOR(Ser2448) by ∼30% above the values observed in the control (no amino acid ingestion) studies; p-AKT(Ser473) and p-AKT(Thr308) were not affected by whey protein or leucine ingestion. Whey protein ingestion decreased insulin-mediated glucose disposal (median 38.8 [quartiles 30.8, 61.8] vs. 51.9 [41.0, 77.3] µmol glucose/µU insulin · mL(-1) · min(-1); P < 0.01), whereas ingestion of leucine did not (52.3 [43.3, 65.4] vs. 52.3 [43.9, 73.2]). These results indicate that 1) protein ingestion causes insulin resistance and could be an important regulator of postprandial glucose homeostasis and 2) the insulin-desensitizing effect of protein ingestion is not due to inhibition of AKT by leucine-mediated mTOR signaling.

Trial registration: ClinicalTrials.gov NCT01538836 NCT01757340.

© 2015 by the American Diabetes Association. Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered.

Figures

Figure 1
Figure 1
Effects of whey protein and leucine ingestion on whole-body glucose Rd (upper panel) and leg glucose uptake (lower panel) (□, basal; ■, clamp). Data are means ± SEM. Three-way ANOVA revealed a significant group (whey protein vs. leucine groups) × study (control vs. whey protein or leucine ingestion) × condition (basal vs. clamp) interaction (P < 0.001) for whole-body glucose Rd. ANCOVA with plasma insulin concentration as a covariate revealed a significant study (control vs. whey protein) × condition (basal vs. clamp) interaction (P < 0.05) for whole-body glucose Rd and leg glucose uptake. Tukey post hoc analysis revealed the following significant differences. *Significantly different from corresponding basal value (P < 0.01); †significantly different from corresponding control value (P < 0.01); ‡significantly different from corresponding control value (P < 0.05).
Figure 2
Figure 2
Effect of whey protein ingestion on p-AMPKThr172, p-ACCSer79, p-mTORSer2448, p-p70S6KThr389, p-AKTSer473, and p-AKTThr308 (arbitrary units) in muscle (□, basal; ■, clamp). AMPK, p-ACC, mTOR, and p-p70S6K data are means ± SEM; AKT data were log transformed for ANOVA and are presented as backtransformed geometric means and errors. Three-way ANOVA revealed a significant study (control vs. whey protein or leucine ingestion) × condition (basal vs. clamp) interaction (P < 0.05) for p-mTORSer2448 and p-p70S6KThr389 and a significant main effect of clamp (P < 0.001) for p-AKTSer473 and p-AKTThr308. Tukey post hoc analysis revealed the following significant differences. *Significantly different from corresponding basal value (P < 0.05); †significantly different from corresponding control value (P < 0.05).
Figure 3
Figure 3
Effect of leucine ingestion on p-AMPKThr172, p-ACCSer79, p-mTORSer2448, p-p70S6KThr389, p-AKTSer473, and p-AKTThr308 (arbitrary units) in muscle (□, basal; ■, clamp). AMPK, p-ACC, mTOR, and p-p70S6K data are means ± SEM; AKT data were log transformed for ANOVA and are presented as backtransformed geometric means and errors. Three-way ANOVA revealed a significant study (control vs. whey protein or leucine ingestion) × condition (basal vs. clamp) interaction (P < 0.05) for p-mTORSer2448 and p-p70S6KThr389 and a significant main effect of clamp (P < 0.001) for p-AKTSer473 and p-AKTThr308. Tukey post hoc analysis revealed the following significant differences. *Significantly different from corresponding basal value (P < 0.05); †significantly different from corresponding control value (P < 0.05).

References

    1. Prager R, Wallace P, Olefsky JM. Hyperinsulinemia does not compensate for peripheral insulin resistance in obesity. Diabetes 1987;36:327–334
    1. Bornfeldt KE, Tabas I. Insulin resistance, hyperglycemia, and atherosclerosis. Cell Metab 2011;14:575–585
    1. Bastien M, Poirier P, Lemieux I, Després JP. Overview of epidemiology and contribution of obesity to cardiovascular disease. Prog Cardiovasc Dis 2014;56:369–381
    1. Newgard CB. Interplay between lipids and branched-chain amino acids in development of insulin resistance. Cell Metab 2012;15:606–614
    1. Adeva MM, Calviño J, Souto G, Donapetry C. Insulin resistance and the metabolism of branched-chain amino acids in humans. Amino Acids 2012;43:171–181
    1. Tremblay F, Jacques H, Marette A. Modulation of insulin action by dietary proteins and amino acids: role of the mammalian target of rapamycin nutrient sensing pathway. Curr Opin Clin Nutr Metab Care 2005;8:457–462
    1. Saha AK, Xu XJ, Balon TW, Brandon A, Kraegen EW, Ruderman NB. Insulin resistance due to nutrient excess: is it a consequence of AMPK downregulation? Cell Cycle 2011;10:3447–3451
    1. Tremblay F, Lavigne C, Jacques H, Marette A. Role of dietary proteins and amino acids in the pathogenesis of insulin resistance. Annu Rev Nutr 2007;27:293–310
    1. Newgard CB, An J, Bain JR, et al. . A branched-chain amino acid-related metabolic signature that differentiates obese and lean humans and contributes to insulin resistance. Cell Metab 2009;9:311–326
    1. Felig P, Marliss E, Cahill GF Jr. Plasma amino acid levels and insulin secretion in obesity. N Engl J Med 1969;281:811–816
    1. Xu F, Tavintharan S, Sum CF, Woon K, Lim SC, Ong CN. Metabolic signature shift in type 2 diabetes mellitus revealed by mass spectrometry-based metabolomics. J Clin Endocrinol Metab 2013;98:E1060–E1065
    1. Würtz P, Soininen P, Kangas AJ, et al. . Branched-chain and aromatic amino acids are predictors of insulin resistance in young adults. Diabetes Care 2013;36:648–655
    1. Menni C, Fauman E, Erte I, et al. . Biomarkers for type 2 diabetes and impaired fasting glucose using a nontargeted metabolomics approach. Diabetes 2013;62:4270–4276
    1. Thalacker-Mercer AE, Ingram KH, Guo F, Ilkayeva O, Newgard CB, Garvey WT. BMI, RQ, diabetes, and sex affect the relationships between amino acids and clamp measures of insulin action in humans. Diabetes 2014;63:791–800
    1. Tzatsos A, Kandror KV. Nutrients suppress phosphatidylinositol 3-kinase/Akt signaling via raptor-dependent mTOR-mediated insulin receptor substrate 1 phosphorylation. Mol Cell Biol 2006;26:63–76
    1. Iwanaka N, Egawa T, Satoubu N, et al. Leucine modulates contraction- and insulin-stimulated glucose transport and upstream signaling events in rat skeletal muscle. J Appl Physiol (1985) 2010;108:274–282
    1. Atherton PJ, Smith K, Etheridge T, Rankin D, Rennie MJ. Distinct anabolic signalling responses to amino acids in C2C12 skeletal muscle cells. Amino Acids 2010;38:1533–1539
    1. Patti ME, Brambilla E, Luzi L, Landaker EJ, Kahn CR. Bidirectional modulation of insulin action by amino acids. J Clin Invest 1998;101:1519–1529
    1. Anthony JC, Yoshizawa F, Anthony TG, Vary TC, Jefferson LS, Kimball SR. Leucine stimulates translation initiation in skeletal muscle of postabsorptive rats via a rapamycin-sensitive pathway. J Nutr 2000;130:2413–2419
    1. Krebs M, Krssak M, Bernroider E, et al. . Mechanism of amino acid-induced skeletal muscle insulin resistance in humans. Diabetes 2002;51:599–605
    1. Flakoll PJ, Wentzel LS, Rice DE, Hill JO, Abumrad NN. Short-term regulation of insulin-mediated glucose utilization in four-day fasted human volunteers: role of amino acid availability. Diabetologia 1992;35:357–366
    1. Pisters PW, Restifo NP, Cersosimo E, Brennan MF. The effects of euglycemic hyperinsulinemia and amino acid infusion on regional and whole body glucose disposal in man. Metabolism 1991;40:59–65
    1. Tremblay F, Krebs M, Dombrowski L, et al. . Overactivation of S6 kinase 1 as a cause of human insulin resistance during increased amino acid availability. Diabetes 2005;54:2674–2684
    1. el-Khoury AE, Sánchez M, Fukagawa NK, Gleason RE, Tsay RH, Young VR. The 24-h kinetics of leucine oxidation in healthy adults receiving a generous leucine intake via three discrete meals. Am J Clin Nutr 1995;62:579–590
    1. Kiskini A, Hamer HM, Wall BT, et al. . The muscle protein synthetic response to the combined ingestion of protein and carbohydrate is not impaired in healthy older men. Age (Dordr) 2013;35:2389–2398
    1. Boirie Y, Gachon P, Corny S, Fauquant J, Maubois JL, Beaufrère B. Acute postprandial changes in leucine metabolism as assessed with an intrinsically labeled milk protein. Am J Physiol 1996;271:E1083–E1091
    1. Gropper SS, Gropper DM, Acosta PB. Plasma amino acid response to ingestion of L-amino acids and whole protein. J Pediatr Gastroenterol Nutr 1993;16:143–150
    1. Rådegran G, Saltin B. Nitric oxide in the regulation of vasomotor tone in human skeletal muscle. Am J Physiol 1999;276:H1951–H1960
    1. Smith GI, Villareal DT, Mittendorfer B. Measurement of human mixed muscle protein fractional synthesis rate depends on the choice of amino acid tracer. Am J Physiol Endocrinol Metab 2007;293:E666–E671
    1. Yoshino J, Conte C, Fontana L, et al. . Resveratrol supplementation does not improve metabolic function in nonobese women with normal glucose tolerance. Cell Metab 2012;16:658–664
    1. Saha AK, Xu XJ, Lawson E, et al. . Downregulation of AMPK accompanies leucine- and glucose-induced increases in protein synthesis and insulin resistance in rat skeletal muscle. Diabetes 2010;59:2426–2434
    1. Goodman CA, Frey JW, Mabrey DM, et al. . The role of skeletal muscle mTOR in the regulation of mechanical load-induced growth. J Physiol 2011;589:5485–5501
    1. Livingstone M, Bidinosti M. Rapamycin-insensitive mTORC1 activity controls eIF4E:4E-BP1 binding. F1000 Res 2012;1:4.
    1. Huo Y, Iadevaia V, Yao Z, et al. . Stable isotope-labelling analysis of the impact of inhibition of the mammalian target of rapamycin on protein synthesis. Biochem J 2012;444:141–151
    1. Greiwe JS, Kwon G, McDaniel ML, Semenkovich CF. Leucine and insulin activate p70 S6 kinase through different pathways in human skeletal muscle. Am J Physiol Endocrinol Metab 2001;281:E466–E471
    1. Baron AD, Brechtel G, Wallace P, Edelman SV. Rates and tissue sites of non-insulin- and insulin-mediated glucose uptake in humans. Am J Physiol 1988;255:E769–E774
    1. Edelman SV, Laakso M, Wallace P, Brechtel G, Olefsky JM, Baron AD. Kinetics of insulin-mediated and non-insulin-mediated glucose uptake in humans. Diabetes 1990;39:955–964
    1. Pruznak AM, Kazi AA, Frost RA, Vary TC, Lang CH. Activation of AMP-activated protein kinase by 5-aminoimidazole-4-carboxamide-1-beta-D-ribonucleoside prevents leucine-stimulated protein synthesis in rat skeletal muscle. J Nutr 2008;138:1887–1894
    1. Wilson GJ, Layman DK, Moulton CJ, et al. . Leucine or carbohydrate supplementation reduces AMPK and eEF2 phosphorylation and extends postprandial muscle protein synthesis in rats. Am J Physiol Endocrinol Metab 2011;301:E1236–E1242
    1. Høeg LD, Sjøberg KA, Lundsgaard AM, et al. . Adiponectin concentration is associated with muscle insulin sensitivity, AMPK phosphorylation, and ceramide content in skeletal muscles of men but not women. J Appl Physiol (1985) 2013;114:592–601
    1. Atherton PJ, Etheridge T, Watt PW, et al. . Muscle full effect after oral protein: time-dependent concordance and discordance between human muscle protein synthesis and mTORC1 signaling. Am J Clin Nutr 2010;92:1080–1088
    1. Højlund K, Glintborg D, Andersen NR, et al. . Impaired insulin-stimulated phosphorylation of Akt and AS160 in skeletal muscle of women with polycystic ovary syndrome is reversed by pioglitazone treatment. Diabetes 2008;57:357–366
    1. Vendelbo MH, Clasen BF, Treebak JT, et al. . Insulin resistance after a 72-h fast is associated with impaired AS160 phosphorylation and accumulation of lipid and glycogen in human skeletal muscle. Am J Physiol Endocrinol Metab 2012;302:E190–E200
    1. Dodd KM, Tee AR. Leucine and mTORC1: a complex relationship. Am J Physiol Endocrinol Metab 2012;302:E1329–E1342
    1. Proud CG. Amino acids and mTOR signalling in anabolic function. Biochem Soc Trans 2007;35:1187–1190
    1. Lang CH, Frost RA, Vary TC. Regulation of muscle protein synthesis during sepsis and inflammation. Am J Physiol Endocrinol Metab 2007;293:E453–E459
    1. Avruch J, Long X, Ortiz-Vega S, Rapley J, Papageorgiou A, Dai N. Amino acid regulation of TOR complex 1. Am J Physiol Endocrinol Metab 2009;296:E592–E602
    1. Conte C, Fabbrini E, Kars M, Mittendorfer B, Patterson BW, Klein S. Multiorgan insulin sensitivity in lean and obese subjects. Diabetes Care 2012;35:1316–1321
    1. Boden G, Tappy L. Effects of amino acids on glucose disposal. Diabetes 1990;39:1079–1084
    1. Ranawana V, Kaur B. Role of proteins in insulin secretion and glycemic control. Adv Food Nutr Res 2013;70:1–47

Source: PubMed

Sponsor e collaboratori

Condizioni mediche

Interventi farmacologici

3
Sottoscrivi