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Cocoa polyphenols suppress adipogenesis in vitro and obesity in vivo by targeting insulin receptor

International Journal of Obesity volume 37, pages 584–592 (2013)Cite this article

Subjects

Abstract

Objective:

To investigate the inhibitory effect of cocoa polyphenol extract (CPE) on adipogenesis and obesity along with its mechanism of action.

Methods and Results:

3T3-L1 preadipocytes were cultured with isobutylmethylxanthine, dexamethasone and insulin (MDI), and male C57BL/6N mice (N=44) were fed a high-fat diet (HFD) for 5 weeks with or without CPE. CPE at 100 or 200 μg ml−1 inhibited MDI-induced lipid accumulation without diminishing cell viability. In particular, CPE reduced the protein expression levels of PPARγ and CEBPα, and blocked mitotic clonal expansion (MCE) of preadipocytes by reducing proliferating signaling pathways. This in turn attenuates lipid accumulation during the differentiation of 3T3-L1 preadipocytes. CPE effectively suppressed MDI-induced phosphorylation of extracellular signal-regulated kinase (ERK) and Akt, and their downstream signals. We then examined whether CPE regulates insulin receptor (IR), a common upstream regulator of ERK and Akt. We found that although CPE does not affect the protein expression level of IR, it significantly inhibits the activity of IR kinase via direct binding. Collectively, the results suggested that CPE, a direct inhibitor of IR kinase activity, inhibits cellular differentiation and lipid accumulation in 3T3-L1 preadipocytes. Consistently, CPE attenuated HFD-induced body weight gain and fat accumulation in obese mice fed with a HFD. We also found that HFD-induced increased fasting glucose levels remained unaffected by CPE.

Conclusion:

This study demonstrates that CPE inhibits IR kinase activity and its proliferative downstream signaling markers, such as ERK and Akt, in 3T3-L1 preadipocytes, and also prevents the development of obesity in mice fed with a HFD.

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References

  1. Hotamisligil G . Inflammation and metabolic disorders. Nature 2006; 444: 860–867.

    Article  CAS  PubMed  Google Scholar 

  2. Kopelman P . Obesity as a medical problem. Nature 2000; 404: 635–643.

    Article  CAS  PubMed  Google Scholar 

  3. Shepherd PR, Gnudi L, Tozzo E, Yang H, Leach F, Kahn BB . Adipose cell hyperplasia and enhanced glucose disposal in transgenic mice overexpressing GLUT4 selectively in adipose tissue. J Biol Chem 1993; 268: 22243–22246.

    CAS  PubMed  Google Scholar 

  4. Naaz A, Holsberger D, Iwamoto G, Nelson A, Kiyokawa H, Cooke P . Loss of cyclin-dependent kinase inhibitors produces adipocyte hyperplasia and obesity. FASEB J 2004; 18: 1925–1927.

    Article  CAS  PubMed  Google Scholar 

  5. Lobstein T, Baur L, Uauy R . Obesity in children and young people: a crisis in public health. Obes Rev 2004; 5 (s1): 4–85.

    Article  PubMed  Google Scholar 

  6. Macdougald OA, Lane MD . Transcriptional regulation of gene-expression during adipocyte differentiation. Annu Rev Biochem 1995; 64: 345–373.

    Article  CAS  PubMed  Google Scholar 

  7. Lane MD, Tang QQ . From multipotent stem cell to adipocyte. Birth Defects Res 2005; 73: 476–477.

    Article  CAS  Google Scholar 

  8. Tang Q-Q, Otto TC, Lane MD . Mitotic clonal expansion: a synchronous process required for adipogenesis. Proc Natl Acad Sci USA 2003; 100: 44–49.

    Article  CAS  PubMed  Google Scholar 

  9. Boulton TG, Nye SH, Robbins DJ, Ip NY, Radzlejewska E, Morgenbesser SD et al. ERKs: a family of protein-serine/threonine kinases that are activated and tyrosine phosphorylated in response to insulin and NGF. Cell 1991; 65: 663–675.

    Article  CAS  PubMed  Google Scholar 

  10. Prusty D, Park B, Davis K, Farmer S . Activation of MEK/ERK signaling promotes adipogenesis by enhancing peroxisome proliferator-activated receptor γ (PPARγ) and C/EBPα gene expression during the differentiation of 3T3-L1 preadipocytes. J Biol Chem 2002; 277: 46226.

    Article  CAS  PubMed  Google Scholar 

  11. Bost F, Aouadi M, Caron L, Binetruy B . The role of MAPKs in adipocyte differentiation and obesity. Biochimie 2005; 87: 51–56.

    Article  CAS  PubMed  Google Scholar 

  12. Taniguchi C, Emanuelli B, Kahn C . Critical nodes in signalling pathways: insights into insulin action. Nat Rev Mol Cell Biol 2006; 7: 85–96.

    Article  CAS  PubMed  Google Scholar 

  13. Tang QQ, Gronborg M, Huang H, Kim JW, Otto TC, Pandey A et al. Sequential phosphorylation of CCAAT enhancer-binding protein beta by MAPK and glycogen synthase kinase 3beta is required for adipogenesis. Proc Natl Acad Sci USA 2005; 102: 9766–9771.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Miyaoka Y, Tanaka M, Naiki T, Miyajima A . Oncostatin M inhibits adipogenesis through the RAS/ERK and STAT5 signaling pathways. J Biol Chem 2006; 281: 37913–37920.

    Article  CAS  PubMed  Google Scholar 

  15. Li X, Kim JW, Gronborg M, Urlaub H, Lane MD, Tang QQ . Role of cdk2 in the sequential phosphorylation/activation of C/EBPbeta during adipocyte differentiation. Proc Natl Acad Sci USA 2007; 104: 11597–11602.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Xu J, Liao K . Protein kinase B/AKT 1 plays a pivotal role in insulin-like growth factor-1 receptor signaling induced 3T3-L1 adipocyte differentiation. J Biol Chem 2004; 279: 35914.

    Article  CAS  PubMed  Google Scholar 

  17. Hu E, Kim J, Sarraf P, Bruce M . Inhibition of adipogenesis through MAP kinase-mediated phosphorylation of PPARgamma. Science 1996; 274: 2100.

    Article  CAS  PubMed  Google Scholar 

  18. White MF, Shoelson SE, Keutmann H, Kahn CR . A cascade of tyrosine autophosphorylation in the beta-subunit activates the phosphotransferase of the insulin receptor. J Biol Chem 1988; 263: 2969–2980.

    CAS  PubMed  Google Scholar 

  19. Saltiel AR, Kahn CR . Insulin signalling and the regulation of glucose and lipid metabolism. Nature 2001; 414: 799–806.

    Article  CAS  PubMed  Google Scholar 

  20. Miki H, Yamauchi T, Suzuki R, Komeda K, Tsuchida A, Kubota N et al. Essential role of insulin receptor substrate 1 (IRS-1) and IRS-2 in adipocyte differentiation. Mol Cell Biol 2001; 21: 2521.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Bluher M, Michael M, Peroni O, Ueki K, Carter N, Kahn B et al. Adipose tissue selective insulin receptor knockout protects against obesity and obesity-related glucose intolerance. Dev Cell 2002; 3: 25–38.

    Article  CAS  PubMed  Google Scholar 

  22. Lee K, Kim Y, Lee H, Lee C . Cocoa has more phenolic phytochemicals and a higher antioxidant capacity than teas and red wine. J Agric Food Chem 2003; 51: 7292–7295.

    Article  CAS  PubMed  Google Scholar 

  23. Ruzaidi A, Amin I, Nawalyah A, Hamid M, Faizul H . The effect of Malaysian cocoa extract on glucose levels and lipid profiles in diabetic rats. J Ethnopharmacol 2005; 98: 55–60.

    Article  CAS  PubMed  Google Scholar 

  24. Lamuela-Raventos R, Romero-Perez A, Andres-Lacueva C, Tornero A Health effects of cocoa flavonoids. Food Sci Technol Int 2005; 11: 159.

  25. Taubert D, Roesen R, Lehmann C, Jung N, Schomig E . Effects of low habitual cocoa intake on blood pressure and bioactive nitric oxide: a randomized controlled trial. JAMA 2007; 298: 49.

    Article  CAS  PubMed  Google Scholar 

  26. Kang N, Lee K, Lee D, Rogozin E, Bode A, Lee H et al. Cocoa procyanidins suppress transformation by inhibiting mitogen-activated protein kinase kinase. J Biol Chem 2008; 283: 20664.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Balzer J, Rassaf T, Heiss C, Kleinbongard P, Lauer T, Merx M et al. Sustained benefits in vascular function through flavanol-containing cocoa in medicated diabetic patients: a double-masked, randomized, controlled trial. J Am Coll Cardiol 2008; 51: 2141.

    Article  CAS  PubMed  Google Scholar 

  28. Matsui N, Ito R, Nishimura E, Yoshikawa M, Kato M, Kamei M et al. Ingested cocoa can prevent high-fat diet-induced obesity by regulating the expression of genes for fatty acid metabolism. Nutrition 2005; 21: 594–601.

    Article  CAS  PubMed  Google Scholar 

  29. Lee K, Kundu J, Kim S, Chun K, Lee H, Surh Y . Cocoa polyphenols inhibit phorbol ester-Induced superoxide anion formation in cultured HL-60 cells and expression of cyclooxygenase-2 and activation of NF-{kappa} B and MAPKs in mouse skin in vivo. J Nutr 2006; 136: 1150.

    Article  CAS  PubMed  Google Scholar 

  30. Cho ES, Jang YJ, Kang NJ, Hwang MK, Kim YT, Lee KW et al. Cocoa procyanidins attenuate 4-hydroxynonenal-induced apoptosis of PC12 cells by directly inhibiting mitogen-activated protein kinase kinase 4 activity. Free Radic Biol Med 2009; 46: 1319–1327.

    Article  CAS  PubMed  Google Scholar 

  31. Cristancho AG, Lazar MA . Forming functional fat: a growing understanding of adipocyte differentiation. Nat Rev Mol Cell Biol 2011; 12: 722–734.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Zhang W, Liu HT . MAPK signal pathways in the regulation of cell proliferation in mammalian cells. Cell Res 2002; 12: 9–18.

    Article  CAS  PubMed  Google Scholar 

  33. Manning BD, Cantley LC . AKT/PKB signaling: navigating downstream. Cell 2007; 129: 1261–1274.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Avruch J . Insulin signal transduction through protein kinase cascades. Mol Cell Biochem 1998; 182: 31–48.

    Article  CAS  PubMed  Google Scholar 

  35. Taylor S . Lilly Lecture: molecular mechanisms of insulin resistance. Lessons from patients with mutations in the insulin-receptor gene. Diabetes 1992; 41: 1473.

    Article  CAS  PubMed  Google Scholar 

  36. Arner P, Pollare T, Lithell H, Livingston J . Defective insulin receptor tyrosine kinase in human skeletal muscle in obesity and type 2 (non-insulin-dependent) diabetes mellitus. Diabetologia 1987; 30: 437–440.

    Article  CAS  PubMed  Google Scholar 

  37. Michael M, Kulkarni R, Postic C, Previs S, Shulman G, Magnuson M et al. Loss of insulin signaling in hepatocytes leads to severe insulin resistance and progressive hepatic dysfunction. Mol Cell 2000; 6: 87–97.

    Article  CAS  PubMed  Google Scholar 

  38. Grassi D, Necozione S, Lippi C, Croce G, Valeri L, Pasqualetti P et al. Cocoa reduces blood pressure and insulin resistance and improves endothelium-dependent vasodilation in hypertensives. Hypertension 2005; 46: 398–405.

    Article  CAS  PubMed  Google Scholar 

  39. Bruning J, Michael M, Winnay J, Hayashi T, Horsch D, Accili D et al. A muscle-specific insulin receptor knockout exhibits features of the metabolic syndrome of NIDDM without altering glucose tolerance. Mol Cell 1998; 2: 559–569.

    Article  CAS  PubMed  Google Scholar 

  40. Zick Y . Ser/Thr phosphorylation of IRS proteins: a molecular basis for insulin resistance. Science’s STKE 2005; 2005: pe4.

    PubMed  Google Scholar 

  41. Bouzakri K, Roques M, Gual P, Espinosa S, Guebre-Egziabher F, Riou J et al. Reduced activation of phosphatidylinositol-3 kinase and increased serine 636 phosphorylation of insulin receptor substrate-1 in primary culture of skeletal muscle cells from patients with type 2 diabetes. Diabetes 2003; 52: 1319.

    Article  CAS  PubMed  Google Scholar 

  42. Harrington L, Findlay G, Gray A, Tolkacheva T, Wigfield S, Rebholz H et al. The TSC1-2 tumor suppressor controls insulin-PI3K signaling via regulation of IRS proteins. J Cell Biol 2004; 166: 213.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Miller B, Shankavaram U, Horney M, Gore A, Kurtz D, Rosenzweig S . Activation of cJun NH2-terminal kinase/stress-activated protein kinase by insulin†. Biochemistry 1996; 35: 8769–8775.

    Article  CAS  PubMed  Google Scholar 

  44. Hong S, Huo H, Xu J, Liao K . Insulin-like growth factor-1 receptor signaling in 3T3-L1 adipocyte differentiation requires lipid rafts but not caveolae. Cell Death Differ 2004; 11: 714–723.

    Article  CAS  PubMed  Google Scholar 

  45. de la Garza AL, Milagro FI, Boque N, Campion J, Martinez JA . Natural inhibitors of pancreatic lipase as new players in obesity treatment. Planta Medica 2011; 77: 773–785.

    Article  CAS  PubMed  Google Scholar 

  46. Tanaka T, Matsuo Y, Kouno I . Chemistry of secondary polyphenols produced during processing of tea and selected foods. Int J Mol Sci 2009; 11: 14–40.

    Article  PubMed  PubMed Central  Google Scholar 

  47. Gu Y, Hurst WJ, Stuart DA, Lambert JD . Inhibition of key digestive enzymes by cocoa extracts and procyanidins. J Agric Food Chem 2011; 59: 5305–5311.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by grants from the World Class University Program (R31-2008-00-10056-0) and the National Leap Research Program (No. 2010-0029233) through the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology, Republic of Korea.

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Author notes

  1. S Y Min and H Yang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Agricultural Biotechnology, Major in Biomodulation, WCU, Seoul National University, Seoul, Republic of Korea

    S Y Min, H Yang, S G Seo, S H Shin, J Kim, S J Lee & H J Lee

  2. Department of Agricultural Biotechnology, Food Science and Biotechnology Program, Seoul National University, Seoul, Republic of Korea

    S Y Min, H Yang, S G Seo, S H Shin, M-Y Chung, S J Lee & K W Lee

  3. Center for Agricultural Biomaterials, Seoul National University, Seoul, Republic of Korea

    M-Y Chung, J Kim & K W Lee

  4. Advanced Institutes of Convergence Technology, Seoul National University, Seoul, Republic of Korea

    J Kim & K W Lee

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Min, S., Yang, H., Seo, S. et al. Cocoa polyphenols suppress adipogenesis in vitro and obesity in vivo by targeting insulin receptor. Int J Obes 37, 584–592 (2013). https://doi.org/10.1038/ijo.2012.85

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