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Clinical Studies and Practice

Phosphorylation of glycogen synthase kinase-3β in metabolically abnormal obesity affects immune stimulation-induced cytokine production

International Journal of Obesity volume 39, pages 270–278 (2015)Cite this article

Subjects

Abstract

Background:

Obesity is a severe health problem worldwide, which leads to multiple comorbidities including type 2 diabetes mellitus and cardiovascular diseases. Inflammation has been found to be an important characteristic of adipose tissue in obese subjects. However, obesity is also associated with compromised immune responses to infections and the impact of obesity on immune function has not been fully understood.

Subjects/Methods:

To clarify the role of obesity in the immune responses, we investigated the Toll-like receptor (TLR)-induced cytokine secretion by leukocytes from obese and lean subjects. We also investigated the relationship between insulin-induced intracellular signaling and cytokine production using peripheral blood mononuclear cells (PBMCs) and a monocytic cell line THP-1.

Results:

We found decreased TLR-induced interferon-γ, interleukin-6 (IL-6) and tumor necrosis factor-α secretions and elevated IL-10 secretion by leukocytes from obese subjects when compared with lean controls. PBMCs from obese subjects showed enhanced basal Akt and glycogen synthase kinase-3β (GSK-3β) phosphorylation, which did not further increase with insulin and lipopolysaccharide (LPS) stimulation. We also found that LPS-induced IκBα degradation was inhibited in PBMCs from obese subjects. By using THP-1 cells with GSK-3β knockdown or cells treated with hyperinsulinemic and high-fatty acid conditions, we found that LPS-induced nuclear factor κB (NF-κB) activation was inhibited and cyclic adenosine monophosphate response element-binding protein (CREB) activation was enhanced.

Conclusions:

These findings indicate that GSK-3β is important in the regulation of NF-κB and CREB activation in leukocytes under the metabolic condition of obesity. Our study revealed a key mechanism through which metabolic abnormalities compromise leukocyte functions in people with obesity.

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References

  1. Meshkani R, Adeli K . Hepatic insulin resistance, metabolic syndrome and cardiovascular disease. Clin Biochem 2009; 42: 1331–1346.

    Article  CAS  Google Scholar 

  2. Odegaard JI, Chawla A . Pleiotropic actions of insulin resistance and inflammation in metabolic homeostasis. Science 2013; 339: 172–177.

    Article  CAS  Google Scholar 

  3. Bunn RC, Cockrell GE, Ou Y, Thrailkill KM, Lumpkin CK Jr, Fowlkes JL . Palmitate and insulin synergistically induce IL-6 expression in human monocytes. Cardiovasc Diabetol 2010; 9: 73.

    Article  Google Scholar 

  4. Dasu MR, Jialal I . Free fatty acids in the presence of high glucose amplify monocyte inflammation via Toll-like receptors. Am J Physiol Endocrinol Metab 2011; 300: E145–E154.

    Article  CAS  Google Scholar 

  5. Huttunen R, Syrjanen J . Obesity and the risk and outcome of infection. Int J Obes (Lond) 2013; 37: 333–340.

    Article  CAS  Google Scholar 

  6. Karlsson EA, Beck MA . The burden of obesity on infectious disease. Exp Biol Med 2010; 235: 1412–1424.

    Article  CAS  Google Scholar 

  7. Arslan E, Atilgan H, Yavasoglu I . The prevalence of Helicobacter pylori in obese subjects. Eur J of Intern Med 2009; 20: 695–697.

    Article  Google Scholar 

  8. Centers for Disease C, Prevention. Intensive-care patients with severe novel influenza A (H1N1) virus infection - Michigan, June 2009. MMWR Morb Mortal Wkly Rep 2009; 58: 749–752.

    Google Scholar 

  9. Amar S, Zhou Q, Shaik-Dasthagirisaheb Y, Leeman S . Diet-induced obesity in mice causes changes in immune responses and bone loss manifested by bacterial challenge. Proc Natl Acad Sci USA 2007; 104: 20466–20471.

    Article  CAS  Google Scholar 

  10. Strandberg L, Verdrengh M, Enge M, Andersson N, Amu S, Onnheim K et al. Mice chronically fed high-fat diet have increased mortality and disturbed immune response in sepsis. PloS one 2009; 4: e7605.

    Article  Google Scholar 

  11. Chang ZL . Important aspects of Toll-like receptors, ligands and their signaling pathways. Inflamm Res 2010; 59: 791–808.

    Article  CAS  Google Scholar 

  12. Hussain R, Kaleem A, Shahid F, Dojki M, Jamil B, Mehmood H et al. Cytokine profiles using whole-blood assays can discriminate between tuberculosis patients and healthy endemic controls in a BCG-vaccinated population. J Immunol Methods 2002; 264: 95–108.

    Article  CAS  Google Scholar 

  13. Dandona P, Aljada A, Mohanty P, Ghanim H, Hamouda W, Assian E et al. Insulin inhibits intranuclear nuclear factor kappaB and stimulates IkappaB in mononuclear cells in obese subjects: evidence for an anti-inflammatory effect? J Clin Endocrinol Metab 2001; 86: 3257–3265.

    CAS  PubMed  Google Scholar 

  14. Lumeng CN . Innate immune activation in obesity. Mol Aspects Med 2013; 34: 12–29.

    Article  CAS  Google Scholar 

  15. Martin M, Rehani K, Jope RS, Michalek SM . Toll-like receptor-mediated cytokine production is differentially regulated by glycogen synthase kinase 3. Nat Immunol 2005; 6: 777–784.

    Article  CAS  Google Scholar 

  16. Nandan D, Camargo de Oliveira C, Moeenrezakhanlou A, Lopez M, Silverman JM, Subek J et al. Myeloid cell IL-10 production in response to leishmania involves inactivation of glycogen synthase kinase-3beta downstream of phosphatidylinositol-3 kinase. J Immunol 2012; 188: 367–378.

    Article  CAS  Google Scholar 

  17. Eldar-Finkelman H, Schreyer SA, Shinohara MM, LeBoeuf RC, Krebs EG . Increased glycogen synthase kinase-3 activity in diabetes- and obesity-prone C57BL/6J mice. Diabetes 1999; 48: 1662–1666.

    Article  CAS  Google Scholar 

  18. Hsu MC, Chang CS, Lee KT, Sun HY, Tsai YS, Kuo PH et al. Central obesity in males affected by a dyslipidemia-associated genetic polymorphism on APOA1/C3/A4/A5 gene cluster. Nutr Diabetes 2013; 3: e61.

    Article  CAS  Google Scholar 

  19. Feinberg J, Fieschi C, Doffinger R, Feinberg M, Leclerc T, Boisson-Dupuis S et al. Bacillus Calmette Guerin triggers the IL-12/IFN-gamma axis by an IRAK-4- and NEMO-dependent, non-cognate interaction between monocytes, NK, and T lymphocytes. Euro J Immunol 2004; 34: 3276–3284.

    Article  CAS  Google Scholar 

  20. Liu SY, Wang WZ, Yen CL, Tsai MY, Yang PW, Wang JY et al. Leukocyte nicotinamide adenine dinucleotide phosphate-reduced oxidase is required for isocyanate-induced lung inflammation. J Allergy Clin Immunol 2011; 127: 1014–1023.

    Article  CAS  Google Scholar 

  21. Huang WC, Tsai CC, Chen CL, Chen TY, Chen YP, Lin YS et al. Glucosylceramide synthase inhibitor PDMP sensitizes chronic myeloid leukemia T315I mutant to Bcr-Abl inhibitor and cooperatively induces glycogen synthase kinase-3-regulated apoptosis. FASEB J 2011; 25: 3661–3673.

    Article  CAS  Google Scholar 

  22. van Dooren FH, Duijvis NW, te Velde AA . Analysis of cytokines and chemokines produced by whole blood, peripheral mononuclear and polymorphonuclear cells. J Immunol Methods 2013; 396: 128–133.

    Article  CAS  Google Scholar 

  23. Xing L, Remick DG . Relative cytokine and cytokine inhibitor production by mononuclear cells and neutrophils. Shock 2003; 20: 10–16.

    Article  CAS  Google Scholar 

  24. Damsgaard CT, Lauritzen L, Calder PC, Kjaer TM, Frokiaer H . Whole-blood culture is a valid low-cost method to measure monocytic cytokines - a comparison of cytokine production in cultures of human whole-blood, mononuclear cells and monocytes. J Immunol Methods 2009; 340: 95–101.

    Article  CAS  Google Scholar 

  25. Staehr P, Hother-Nielsen O, Landau BR, Chandramouli V, Holst JJ, Beck-Nielsen H . Effects of free fatty acids per se on glucose production, gluconeogenesis, and glycogenolysis. Diabetes 2003; 52: 260–267.

    Article  CAS  Google Scholar 

  26. Wen AY, Sakamoto KM, Miller LS . The role of the transcription factor CREB in immune function. J Immunol 2010; 185: 6413–6419.

    Article  CAS  Google Scholar 

  27. Andersson CX, Gustafson B, Hammarstedt A, Hedjazifar S, Smith U . Inflamed adipose tissue, insulin resistance and vascular injury. Diabetes Metab Res Rev 2008; 24: 595–603.

    Article  CAS  Google Scholar 

  28. Falagas ME, Kompoti M . Obesity and infection. Lancet Infect Dis 2006; 6: 438–446.

    Article  Google Scholar 

  29. Chang JT, Dou HY, Yen CL, Wu YH, Huang RM, Lin HJ et al. Effect of type 2 diabetes mellitus on the clinical severity and treatment outcome in patients with pulmonary tuberculosis: a potential role in the emergence of multidrug-resistance. J Formos Med Assoc 2011; 110: 372–381.

    Article  Google Scholar 

  30. Jain S, Chaves SS . Obesity and influenza. Clin Infect Dis 2011; 53: 422–424.

    Article  Google Scholar 

  31. Gleeson M, Bishop N, Oliveira M, McCauley T, Tauler P, Muhamad AS . Respiratory infection risk in athletes: association with antigen-stimulated IL-10 production and salivary IgA secretion. Scand J Med Sci Sports 2012; 22: 410–417.

    Article  CAS  Google Scholar 

  32. Hill JR, Kwon G, Marshall CA, McDaniel ML . Hyperglycemic levels of glucose inhibit interleukin 1 release from RAW 264.7 murine macrophages by activation of protein kinase C. J Biol Chem 1998; 273: 3308–3313.

    Article  CAS  Google Scholar 

  33. Turina M, Fry DE, Polk HC Jr . Acute hyperglycemia and the innate immune system: clinical, cellular, and molecular aspects. Crit Care Med 2005; 33: 1624–1633.

    Article  Google Scholar 

  34. Kidd LB, Schabbauer GA, Luyendyk JP, Holscher TD, Tilley RE, Tencati M et al. Insulin activation of the phosphatidylinositol 3-kinase/protein kinase B (Akt) pathway reduces lipopolysaccharide-induced inflammation in mice. J Pharmacol Exp Ther 2008; 326: 348–353.

    Article  CAS  Google Scholar 

  35. Cohen P . The Croonian Lecture 1998. Identification of a protein kinase cascade of major importance in insulin signal transduction. Philos Trans R Soc Lond B Biol Sci 1999; 354: 485–495.

    Article  CAS  Google Scholar 

  36. van den Berghe G, Wouters P, Weekers F, Verwaest C, Bruyninckx F, Schetz M et al. Intensive insulin therapy in critically ill patients. N Engl J Med 2001; 345: 1359–1367.

    Article  CAS  Google Scholar 

  37. Zhou Q, Leeman SE, Amar S . Signaling mechanisms involved in altered function of macrophages from diet-induced obese mice affect immune responses. Proc Natl Acad Sci USA 2009; 106: 10740–10745.

    Article  CAS  Google Scholar 

  38. Cremer TJ, Shah P, Cormet-Boyaka E, Valvano MA, Butchar JP, Tridandapani S . Akt-mediated proinflammatory response of mononuclear phagocytes infected with Burkholderia cenocepacia occurs by a novel GSK3beta-dependent, IkappaB kinase-independent mechanism. J Immunol 2011; 187: 635–643.

    Article  CAS  Google Scholar 

  39. Hansen T, Rehfeld JF, Nielsen FC . GSK-3beta reduces cAMP-induced cholecystokinin gene expression by inhibiting CREB binding. Neuroreport 2004; 15: 841–845.

    Article  CAS  Google Scholar 

  40. Ahmad R, Al-Mass A, Atizado V, Al-Hubail A, Al-Ghimlas F, Al-Arouj M et al. Elevated expression of the toll like receptors 2 and 4 in obese individuals: its significance for obesity-induced inflammation. J Inflamm 2012; 9: 48.

    Article  CAS  Google Scholar 

  41. Galic S, Oakhill JS, Steinberg GR . Adipose tissue as an endocrine organ. Mol Cell Biol 2010; 316: 129–139.

    CAS  Google Scholar 

  42. Fang X, Yu S, Tanyi JL, Lu Y, Woodgett JR, Mills GB . Convergence of multiple signaling cascades at glycogen synthase kinase 3: Edg receptor-mediated phosphorylation and inactivation by lysophosphatidic acid through a protein kinase C-dependent intracellular pathway. Mol Cell Biol 2002; 22: 2099–2110.

    Article  CAS  Google Scholar 

  43. Quan J, Liu J, Gao X, Liu J, Yang H, Chen W et al. Palmitate induces interleukin-8 expression in human aortic vascular smooth muscle cells via Toll-like receptor 4/nuclear factor-kappaB pathway (TLR4/NF-kappaB-8). J Diabetes 2014; 6: 33–41.

    Article  CAS  Google Scholar 

  44. Ban JO, Kwak DH, Oh JH, Park EJ, Cho MC, Song HS et al. Suppression of NF-kappaB and GSK-3beta is involved in colon cancer cell growth inhibition by the PPAR agonist troglitazone. Chem Biol Interact 2010; 188: 75–85.

    Article  CAS  Google Scholar 

  45. Lee S, Yang WK, Song JH, Ra YM, Jeong JH, Choe W et al. Anti-obesity effects of 3-hydroxychromone derivative, a novel small-molecule inhibitor of glycogen synthase kinase-3. Biochem Pharmacol 2013; 85: 965–976.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Professor S-J Tsai, Professor Y-S Lin and Professor T-M Yeh for helpful discussion. This work was supported by National Cheng Kung University Hospital grant (NCKUH9801002).

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Authors and Affiliations

  1. Institute of Basic Medical Science, National Cheng Kung University Medical College, Tainan, Taiwan

    C-L Yen & C-C Shieh

  2. Institute of Clinical Medicine, National Cheng Kung University Medical College, Tainan, Taiwan

    W-C Chao, Y-F Huang, Y-S Tsai, C-F Lin & C-C Shieh

  3. Department of Internal Medicine, Taichung Veteran General Hospital, Chiayi Branch, Chiayi, Taiwan

    W-C Chao

  4. Department of Family Medicine, National Cheng Kung University Hospital, Tainan, Taiwan

    C-H Wu & C-S Chang

  5. Department of Pediatrics, National Cheng Kung University Hospital, Tainan, Taiwan

    C-C Shieh

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Yen, CL., Chao, WC., Wu, CH. et al. Phosphorylation of glycogen synthase kinase-3β in metabolically abnormal obesity affects immune stimulation-induced cytokine production. Int J Obes 39, 270–278 (2015). https://doi.org/10.1038/ijo.2014.93

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