Acute physical exercise reverses S-nitrosation of the insulin receptor, insulin receptor substrate 1 and protein kinase B/akt in diet-induced obese wistar rats

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOEarly evidence demonstrates that exogenous nitric oxide (NO) and the NO produced by inducible nitric oxide synthase (iNOS) can induce insulin resistance. Here, we investigated whether this insulin resistance, mediated by S-nitrosation of proteins involved in early steps of the insulin signal transduction pathway, could be reversed by acute physical exercise. Rats on a high-fat diet were subjected to swimming for two 3 h-long bouts, separated by a 45 min rest period. Two or 16 h after the exercise protocol the rats were killed and proteins from the insulin signalling pathway were analysed by immunoprecipitation and immunoblotting. We demonstrated that a high-fat diet led to an increase in the iNOS protein level and S-nitrosation of insulin receptor β (IRβ), insulin receptor substrate 1 (IRS1) and Akt. Interestingly, an acute bout of exercise reduced iNOS expression and S-nitrosation of proteins involved in the early steps of insulin action, and improved insulin sensitivity in diet-induced obesity rats. Furthermore, administration of GSNO (NO donor) prevents this improvement in insulin action and the use of an inhibitor of iNOS (L-N 6 -(1-iminoethyl)lysine; L-NIL) simulates the effects of exercise on insulin action, insulin signalling and S-nitrosation of IRβ, IRS1 and Akt. In summary, a single bout of exercise reverses insulin sensitivity in diet-induced obese rats by improving the insulin signalling pathway, in parallel with a decrease in iNOS expression and in the S-nitrosation of IR/IRS1/Akt. The decrease in iNOS protein expression in the muscle of diet-induced obese rats after an acute bout of exercise was accompanied by an increase in AMP-activated protein kinase (AMPK) activity. These results provide new insights into the mechanism by which exercise restores insulin sensitivity. © 2008 The Authors. Journal compilation © 2008 The Physiological Society.Early evidence demonstrates that exogenous nitric oxide (NO) and the NO produced by inducible nitric oxide synthase (iNOS) can induce insulin resistance. Here, we investigated whether this insulin resistance, mediated by S-nitrosation of proteins involved in early steps of the insulin signal transduction pathway, could be reversed by acute physical exercise. Rats on a high-fat diet were subjected to swimming for two 3 h-long bouts, separated by a 45 min rest period. Two or 16 h after the exercise protocol the rats were killed and proteins from the insulin signalling pathway were analysed by immunoprecipitation and immunoblotting. We demonstrated that a high-fat diet led to an increase in the iNOS protein level and S-nitrosation of insulin receptor beta (IR beta), insulin receptor substrate 1 (IRS1) and Akt. Interestingly, an acute bout of exercise reduced iNOS expression and S-nitrosation of proteins involved in the early steps of insulin action, and improved insulin sensitivity in diet-induced obesity rats. Furthermore, administration of GSNO (NO donor) prevents this improvement in insulin action and the use of an inhibitor of iNOS (L-N6-(1-iminoethyl)lysine; L-NIL) simulates the effects of exercise on insulin action, insulin signalling and S-nitrosation of IR beta, IRS1 and Akt. In summary, a single bout of exercise reverses insulin sensitivity in diet-induced obese rats by improving the insulin signalling pathway, in parallel with a decrease in iNOS expression and in the S-nitrosation of IR/IRS1/Akt. The decrease in iNOS protein expression in the muscle of diet-induced obese rats after an acute bout of exercise was accompanied by an increase in AMP-activated protein kinase (AMPK) activity. These results provide new insights into the mechanism by which exercise restores insulin sensitivity5862659671FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOArias, E.B., Kim, J., Funai, K., Cartee, G.D., Prior exercise increases phosphorylation of Akt substrate of 160 kDa (AS160) in rat skeletal muscle (2007) Am J Physiol Endocrinol Metab, 292, pp. E1191-E1200Balon, T.W., Nadler, J.L., Evidence that nitric oxide increases glucose transport in skeletal muscle (1997) J Appl Physiol, 82, pp. 359-363Bedard, S., Marcotte, B., Marette, A., Cytokines modulate glucose transport in skeletal muscle by inducing the expression of inducible nitric oxide synthase (1997) Biochem J, 325, pp. 487-493Betts, J.J., Sherman, W.M., Reed, M.J., Gao, J.P., Duration of improved muscle glucose uptake after acute exercise in obese Zucker rats (1993) Obes Res, 1, pp. 295-302Bisquolo, V.A., Cardoso, C.G., Ortega, K.C., Gusmao, J.L., Tinucci, T., Negrao, C.E., Wajchenberg, B.L., Forjaz, C.L., Previous exercise attenuates muscle sympathetic activity and increases blood flow during acute euglycemic hyperinsulinemia (2005) J Appl Physiol, 98, pp. 866-871Bonora, E., Moghetti, P., Zancanaro, C., Cigolini, M., Querena, M., Cacciatori, V., Corgnati, A., Muggeo, M., Estimates of in vivo insulin action in man: Comparison of insulin tolerance tests with euglycemic and hyperglycemic glucose clamp studies (1989) J Clin Endocrinol Metab, 68, pp. 374-378Broillet, M.C., Firestein, S., Direct activation of the olfactory cyclic nucleotide-gated channel through modification of sulfhydryl groups by NO compounds (1996) Neuron, 16, pp. 377-385Bruce, C.R., Lee, J.S., Hawley, J.A., Postexercise muscle glycogen resynthesis in obese insulin-resistant Zucker rats (2001) J Appl Physiol, 91, pp. 1512-1519Carvalho-Filho, M.A., Ueno, M., Carvalheira, J.B., Velloso, L.A., Saad, M.J., Targeted disruption of iNOS prevents LPS-induced S-nitrosation of IRbeta/ IRS-1 and Akt and insulin resistance in muscle of mice (2006) Am J Physiol Endocrinol Metab, 291, pp. E476-E482Carvalho-Filho, M.A., Ueno, M., Hirabara, S.M., Seabra, A.B., Carvalheira, J.B., de Oliveira, M.G., Velloso, L.A., Saad, M.J., S-nitrosation of the insulin receptor, insulin receptor substrate 1, and protein kinase B/Akt: A novel mechanism of insulin resistance (2005) Diabetes, 54, pp. 959-967Chibalin, A.V.Yu.M., Ryder, J.W., Song, X.M., Galuska, D., Krook, A., Wallberg-Henriksson, H., Zierath, J.R., Exercise-induced changes in expression and activity of proteins involved in insulin signal transduction in skeletal muscle: Differential effects on insulin-receptor substrates 1 and 2 (2000) Proc Natl Acad Sci USA, 97, pp. 38-43Devlin, J.T., Hirshman, M., Horton, E.D., Horton, E.S., Enhanced peripheral and splanchnic insulin sensitivity in NIDDM men after single bout exercise (1987) Diabetes, 36, pp. 434-439Elchebly, M., Payette, P., Michaliszyn, E., Cromlish, W., Collins, S., Loy, A.L., Normandin, D., Kennedy, B.P., Increased insulin sensitivity and obesity resistance in mice lacking the protein tyrosine phosphatase-1B gene (1999) Science, 283, pp. 1544-1548Fiedler, M., Zierath, J.R., Selen, G., Wallberg-Henriksson, H., Liang, Y., Sakariassen, K.S., 5-aminoimidazole-4-carboxy-amide-1-beta-D-ribofuranoside treatment ameliorates hyperglycaemia and hyperinsulinaemia but not dyslipidaemia in KKAy-CETP mice (2001) Diabetologia, 44, pp. 2180-2186Gielen, S., Adams, V., Mobius-Winkler, S., Linke, A., Erbs, S.Yu.J., Kempf, W., Schubert, A., Hambrecht, R., Anti-inflammatory effects of exercise training in the skeletal muscle of patients with chronic heart failure (2003) J Am Coll Cardiol, 42, pp. 861-868Gross, S.S., Wolin, M.S., Nitric oxide: Pathophysiological mechanisms (1995) Annu Rev Physiol, 57, pp. 737-769Hirosumi, J., Tuncman, G., Chang, L., Gorgun, C.Z., Uysal, K.T., Maeda, K., Karin, M., Hotamisligil, G.S., A central role for JNK in obesity and insulin resistance (2002) Nature, 420, pp. 333-336Hotamisligil, G.S., Peraldi, P., Budavari, A., Ellis, R., White, M.F., Spiegelman, B.M., IRS-1-mediated inhibition of insulin receptor tyrosine kinase activity in TNF-alpha- and obesity-induced insulin resistance (1996) Science, 271, pp. 665-668Jaffrey, S.R., Snyder, S.H., The biotin switch method for the detection of S-nitrosylated proteins (2001) Sci STKE, 2001, pp. PL1Kapur, S., Bedard, S., Marcotte, B., Cote, C.H., Marette, A., Expression of nitric oxide synthase in skeletal muscle: A novel role for nitric oxide as a modulator of insulin action (1997) Diabetes, 46, pp. 1691-1700Kapur, S., Marcotte, B., Marette, A., Mechanism of adipose tissue iNOS induction in endotoxemia (1999) Am J Physiol Endocrinol Metab, 276, pp. E635-E641Laemmli, U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T4 (1970) Nature, 227, pp. 680-685Lancaster, G.I., Khan, Q., Drysdale, P., Wallace, F., Jeukendrup, A.E., Drayson, M.T., Gleeson, M., The physiological regulation of toll-like receptor expression and function in humans (2005) J Physiol, 563, pp. 945-955Lander, H.M., Ogiste, J.S., Pearce, S.F., Levi, R., Novogrodsky, A., Nitric oxide-stimulated guanine nucleotide exchange on p21ras (1995) J Biol Chem, 270, pp. 7017-7020Lane, P., Gross, S.S., Cell signaling by nitric oxide (1999) Semin Nephrol, 19, pp. 215-229Liu, L., Zhang, Y., Chen, N., Shi, X., Tsang, B., Yu, Y.H., Upregulation of myocellular DGAT1 augments triglyceride synthesis in skeletal muscle and protects against fat-induced insulin resistance (2007) J Clin Invest, 117, pp. 1679-1689Martinez-Ruiz, A., Lamas, S., Detection and proteomic identification of S-nitrosylated proteins in endothelial cells (2004) Arch Biochem Biophys, 423, pp. 192-199McGrowder, D., Ragoobirsingh, D., Brown, P., Modulation of glucose uptake in adipose tissue by nitric oxide-generating compounds (2006) J Biosci, 31, pp. 347-354Molina y Vedia, L., McDonald, B., Reep, B., Brune, B., Di Silvio, M., Billiar, T.R., Lapetina, E.G., Nitric oxide-induced S-nitrosylation of glyceraldehyde-3-phosphate dehydrogenase inhibits enzymatic activity and increases endogenous ADP-ribosylation (1992) J Biol Chem, 267, pp. 24929-24932Perreault, M., Marette, A., Targeted disruption of inducible nitric oxide synthase protects against obesity-linked insulin resistance in muscle (2001) Nat Med, 7, pp. 1138-1143Pilon, G., Dallaire, P., Marette, A., Inhibition of inducible nitric-oxide synthase by activators of AMP-activated protein kinase: A new mechanism of action of insulin-sensitizing drugs (2004) J Biol Chem, 279, pp. 20767-20774Richter, E.A., Garetto, L.P., Goodman, M.N., Ruderman, N.B., Muscle glucose metabolism following exercise in the rat: Increased sensitivity to insulin (1982) J Clin Invest, 69, pp. 785-793Ropelle, E.R., Pauli, J.R., Prada, P.O., de Souza, C.T., Picardi, P.K., Faria, M.C., Cintra, D.E., Carvalheira, J.B., Reversal of diet-induced insulin resistance with a single bout of exercise in the rat: The role of PTP1B and IRS-1 serine phosphorylation (2006) J Physiol, 577, pp. 997-1007Saad, M.J., Maeda, L., Brenelli, S.L., Carvalho, C.R., Paiva, R.S., Velloso, L.A., Defects in insulin signal transduction in liver and muscle of pregnant rats (1997) Diabetologia, 40, pp. 179-186Schenk, S., Horowitz, J.F., Acute exercise increases triglyceride synthesis in skeletal muscle and prevents fatty acid-induced insulin resistance (2007) J Clin Invest, 117, pp. 1690-1698Scott, M.D., Kuypers, F.A., Butikofer, P., Bookchin, R.M., Ortiz, O.E., Lubin, B.H., Effect of osmotic lysis and resealing on red cell structure and function (1990) J Lab Clin Med, 115, pp. 470-480Shi, H., Kokoeva, M.V., Inouye, K., Tzameli, I., Yin, H., Flier, J.S., TLR4 links innate immunity and fatty acid-induced insulin resistance (2006) J Clin Invest, 116, pp. 3015-3025Shishido, S.M., Seabra, A.B., Loh, W., Ganzarolli de Oliveira, M., Thermal and photochemical nitric oxide release from S-nitrosothiols incorporated in Pluronic F127 gel: Potential uses for local and controlled nitric oxide release (2003) Biomaterials, 24, pp. 3543-3553Stamler, J.S., Simon, D.I., Osborne, J.A., Mullins, M.E., Jaraki, O., Michel, T., Singel, D.J., Loscalzo, J., S-nitrosylation of proteins with nitric oxide: Synthesis and characterization of biologically active compounds (1992) Proc Natl Acad Sci USA, 89, pp. 444-448Stamler, J.S., Toone, E.J., Lipton, S.A., Sucher, N.J., (S) NO signals: Translocation, regulation, and a consensus motif (1997) Neuron, 18, pp. 691-696Sugita, H., Fujimoto, M., Yasukawa, T., Shimizu, N., Sugita, M., Yasuhara, S., Martyn, J.A., Kaneki, M., Inducible nitric-oxide synthase and NO donor induce insulin receptor substrate-1 degradation in skeletal muscle cells (2005) J Biol Chem, 280, pp. 14203-14211Tsukumo, D.M., Carvalho-Filho, M.A., Carvalheira, J.B., Prada, P.O., Hirabara, S.M., Schenka, A.A., Araujo, E.P., Saad, M.J., Loss-of-function mutation in Toll-like receptor 4 prevents diet-induced obesity and insulin resistance (2007) Diabetes, 56, pp. 1986-1998Wallberg-Henriksson, H., Glucose transport into skeletal muscle. Influence of contractile activity, insulin, catecholamines and diabetes mellitus (1987) Acta Physiol Scand, 564 (SUPPL.), pp. 1-80Wallberg-Henriksson, H., Constable, S.H., Young, D.A., Holloszy, J.O., Glucose transport into rat skeletal muscle: Interaction between exercise and insulin (1988) J Appl Physiol, 65, pp. 909-913Winder, W.W., AMP-activated protein kinase: Possible target for treatment of type 2 diabetes (2000) Diabetes Technol Ther, 2, pp. 441-448Wojtaszewski, J.F., Birk, J.B., Frosig, C., Holten, M., Pilegaard, H., Dela, F., 5′AMP activated protein kinase expression in human skeletal muscle: Effects of strength training and type 2 diabetes (2005) J Physiol, 564, pp. 563-573Zierath, J.R., In vitro studies of human skeletal muscle: Hormonal and metabolic regulation of glucose transport (1995) Acta Physiol Scand Suppl, 626, pp. 1-9