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Elevated Expression of Transforming Growth Factor-β in Adipose Tissue from Obese Mice

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Abstract

Background

Tumor necrosis factor-α (TNF-α) is chronically elevated in the adipose tissue from obese humans and mice. This increase in TNF-α contributes to the insulin resistance, elevated plasminogen activator inhibitor-1 (PAI-1) levels, and cardiovascular complications associated with obesity and noninsulin-dependent diabetes (NIDDM). PAI-1 gene expression in adipose tissue is also stimulated by transforming growth factor-β (TGF-β). Experiments were performed to determine whether TGF-β is regulated by TNF-α and elevated in obesity.

Materials and Methods

The concentration of TGF-β and PAI-1 mRNA in murine adipose tissue and cultured 3T3-L1 adipocytes was determined by quantitative reverse transcription polymerase chain reaction (RT-PCR), and the cellular localization of these molecules was evaluated using in situ hybridization and cell fractionation. Total TGF-β protein was determined by employing an ELISA assay.

Results

TGF-β mRNA and protein were increased in the adipose tissue from two different strains of genetically obese mice (i.e., ob/ob and db/db), compared with their lean counterparts. This increase in TGF-β may result from TNF-α since TNF-α increased TGF-β mRNA expression in the adipose tissue of lean mice and stimulated TGF-β production by cultured adipocytes. Administration of TGF-β increased PAI-1 antigen in the plasma and PAI-1 mRNA in the adipocytes of lean mice, and enhanced the rate of PAI-1 synthesis by adipocytes in vitro.

Conclusions

TNF-α contributes to the elevated TGF-β expression demonstrated in the adipose tissue of obese mice. A potential role for TGF-β in the increased PAI-1 and vascular pathologies associated with obesity/NIDDM is suggested.

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References

  1. Larsson B. (1991) Obesity, fat distribution and cardiovascular disease. Int. J. Obesity 15: 53–57.

    Google Scholar 

  2. Björntorp P. (1992) Abdominal fat distribution and disease: An overview of epidemiological data. Ann. Med. 24: 15–18.

    Article  PubMed  Google Scholar 

  3. DeFronzo RA, Ferrannini E. (1991) Insulin resistance: A multifaceted syndrome responsible for NIDDM, obesity, hypertension, dyslipidemia, and atherosclerotic cardiovascular disease. Diabetes Care 14: 173–194.

    Article  CAS  PubMed  Google Scholar 

  4. McGill JB, Schneider DJ, Arfken CL, Lucore CL, Sobel BE. (1994) Factors responsible for impaired fibrinolysis in obese subjects and NIDDM patients. Diabetes 43: 104–109.

    Article  CAS  PubMed  Google Scholar 

  5. Potter van Loon BJ, Kluft C, Radder JK, Blankenstein MA, Meinders AE. (1993) The cardiovascular risk factor plasminogen activator inhibitor type 1 is related to insulin resistance. Metabolism 42: 945–949.

    Article  Google Scholar 

  6. Legnani C, Maccaferri M, Tonini P, Cassio A, Cacciari E, Coccheri S. (1988) Reduced fibrinolytic response in obese children: Association with high baseline activity of the fast acting plasminogen activator inhibitor (PAI-1). Fibrinolysis 2: 211–214.

    Article  Google Scholar 

  7. Vague P, Juhan-Vague I, Chabert V, Alessi MC, Atlan C. (1989) Fat distribution and plasminogen activator inhibitor activity in nondiabetic obese women. Metabolism 38: 913–915.

    Article  CAS  PubMed  Google Scholar 

  8. Sprengers ED, Kluft C. (1987) Plasminogen activator inhibitors. Blood 69: 381–387.

    PubMed  CAS  Google Scholar 

  9. Loskutoff DJ. (1988) Type 1 plasminogen activator inhibitor and its potential influence on thrombolytic therapy. Semin. Thromb. Hemost. 14: 100–109.

    Article  CAS  PubMed  Google Scholar 

  10. Schneiderman J, Loskutoff DJ. (1991) Plasminogen activator inhibitors. Trends Cardiovasc. Med. 1: 99–102.

    Article  CAS  PubMed  Google Scholar 

  11. Primrose JN, Davies JA, Prentice CRM, Hughes R, Johnston D. (1992) Reduction in factor VII, fibrinogen and plasminogen activator inhibitor-1 activity after surgical treatment of morbid obesity. Thromb. Haemost. 68: 396–399.

    Article  CAS  PubMed  Google Scholar 

  12. Samad F, Loskutoff DJ. (1996) Tissue distribution and regulation of plasminogen activator inhibitor-1 in obese mice. Mol. Med. 2: 568–582.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Hotamisligil GS, Shargill NS, Spiegelman BM. (1993) Adipose expression of tumor necrosis factor-α: Direct role in obesity-linked insulin resistance. Science 259: 87–91.

    Article  CAS  PubMed  Google Scholar 

  14. Hotamisligil GS, Arner P, Caro JF, Atkinson RL, Spiegelman BM. (1995) Increased adipose tissue expression of tumor necrosis factor-α in human obesity and insulin resistance. J. Clin. Invest. 95: 2409–2415.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. van Hinsbergh VW, Kooistra T, van den Berg EA, Princen HM, Fiers W, Emeis JJ. (1988) Tumor necrosis factor increases the production of plasminogen activator inhibitor in human endothelial cells in vitro and in rats in vivo. Blood 72: 1467–1473.

    PubMed  Google Scholar 

  16. Kooistra T. (1990) The use of cultured human endothelial cells and hepatocytes as an in vitro model system to study modulation of endogenous fibrinolysis. Fibrinolysis 4: 33–39.

    Article  CAS  Google Scholar 

  17. Vague P, Juhan-Vague I, Aillaud MF, Badier C, Viard R, Alessi MC, Collen D. (1986) Correlation between blood fibrinolytic activity, plasminogen activator inhibitor level plasma insulin level, and relative body weight in normal and obese subjects. Metabolism 35: 250–253.

    Article  CAS  PubMed  Google Scholar 

  18. Mykkänen L, Rönnemaa T, Marniemi J, Haffner SM, Bergman R, Laakso M. (1994) Insulin sensitivity is not an independent determinant of plasma plasminogen activator inhibitor-1 activity. Arterioscler. Thromb. 14: 1264–1271.

    Article  PubMed  Google Scholar 

  19. Asplund-Carlson A, Hamsten A, Wiman B, Carlson LA. (1993) Relationship between plasma plasminogen activator inhibitor-1 activity and VLDL triglyceride concentration, insulin levels and insulin sensitivity: Studies in randomly selected normo- and hypertriglyceridaemic men. Diabetologia 36: 817–825.

    Article  CAS  PubMed  Google Scholar 

  20. Samad F, Yamamoto K, Loskutoff DJ. (1996) Distribution and regulation of plasminogen activator inhibitor-1 in murine adipose tissue in vivo: Induction by tumor necrosis factor-α and lipopolysaccharide. J. Clin. Invest. 97: 37–46.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Sawdey MS, Loskutoff DJ. (1991) Regulation of murine type 1 plasminogen activator inhibitor gene expression in vivo: Tissue specificity and induction by lipopoly saccharide, tumor necrosis factor-α, and transforming growth factor-β. J. Clin. Invest. 88: 1346–1353.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Schleef RR, Sinha M, Loskutoff DJ. (1985) Immunoradiometric assay to measure the binding of a specific inhibitor to tissue-type plasminogen activator. J. Lab. Clin. Med. 106: 408–415.

    PubMed  CAS  Google Scholar 

  23. Wang AM, Doyle MV, Mark DF. (1989) Quantitation of mRNA by the polymerase chain reaction. Proc. Natl. Acad. Sci. U.S.A. 86: 9717–9721.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Yamamoto K, Loskutoff DJ. (1996) Fibrin deposition in tissues from endotoxin-treated mice correlates with decreases in the expression of urokinase-type but not tissue-type plasminogen activator. J. Clin. Invest. 97: 2440–2451.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Vanden Heuvel JP, Tyson FL, Bell DA. (1993) Construction of recombinant RNA templates for use as internal standards in quantitative RT-PCR. Bio Techniques 14: 395–398.

    Google Scholar 

  26. Keeton M, Eguchi Y, Sawdey M, Ahn C, Loskutoff DJ. (1993) Cellular localization of type 1 plasminogen activator inhibitor messenger RNA and protein in murine renal tissue. Am. J. Pathol. 142: 59–70.

    PubMed  PubMed Central  CAS  Google Scholar 

  27. Kehrl JH, Wakefield LM, Roberts AB, Jakowlew SB, Alvarez-Mon M, Derynck R, Sporn MB, Fauci AS. (1986) Production of transforming growth factor β by human T lymphocytes and its potential role in the regulation of T cell growth. J. Exp. Med. 163: 1037–1050.

    Article  CAS  PubMed  Google Scholar 

  28. Rodbell M. (1964) Metabolism of isolated fat cells: I. Effects of hormones on glucose metabolism and lipolysis. J. Biol. Chem. 239: 375–380.

    PubMed  CAS  Google Scholar 

  29. Moore DD. (1996) Preparation of genomic DNA from mammalian tissue. In: Ausubel FM, Brent R, Kingston RE, et al. (eds) Current Protocols in Molecular Biology. John Wiley and Sons, New York, Vol. 1, pp. 221–222.

    Google Scholar 

  30. Green H, Kehinde O. (1975) An established preadipose cell line and its differentiation in culture: II. Factors affecting the adipose conversion. Cell 5: 19–27.

    Article  CAS  PubMed  Google Scholar 

  31. Markman B. (1989) Anatomy and physiology of adipose tissue. Clin. Plast. Surg. 16: 235–243.

    PubMed  CAS  Google Scholar 

  32. McCartney-Francis NL, Wahl SM. (1994) Transforming growth factor beta: A matter of life and death. J. Leukocyte Biol. 55: 401–409.

    Article  CAS  PubMed  Google Scholar 

  33. Sporn MB, Roberts AB, Wakefield LM, de Crombrugghe B. (1987) Some recent advances in the chemistry and biology of transforming growth factor-β. J. Cell Biol. 105: 1039–1045.

    Article  CAS  PubMed  Google Scholar 

  34. Assoian RK, Fleurdelys BE, Stevenson HC, Miller PJ, Madtes DK, Raines EW, Ross R, Sporn MB. (1987) Expression and secretion of type beta transforming growth factor by activated human macrophages. Proc. Natl. Acad. Sci. U.S.A. 84: 6020–6024.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Sawdey M, Podor TJ, Loskutoff DJ. (1989) Regulation of type 1 plasminogen activator inhibitor gene expression in cultured bovine aortic endothelial cells: Induction by transforming growth factor-β, lipopolysaccharide, and tumor necrosis factor-α. J. Biol. Chem. 264: 10396–10401.

    PubMed  CAS  Google Scholar 

  36. Reilly CF, McFall RC. (1991) Platelet-derived growth factor and transforming growth factor-β regulate plasminogen activator inhibitor-1 synthesis in vascular smooth muscle cells. J. Biol. Chem. 266: 9419–9427.

    PubMed  CAS  Google Scholar 

  37. Lund LR, Riccio A, Andreasen PA, Nielsen LS, Kristensen P, Laiho M, Saksela O, Blasi F, Dano K. (1987) Transforming growth factor-β is a strong and fast acting positive regulator of the level of type-1 plasminogen activator inhibitor mRNA in WI-38 human lung fibroblasts. EMBO J. 6: 1281–1286.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  38. Thalacker FW, Nilsen-Hamilton M. (1987) Specific induction of secreted proteins by transforming growth factor-β and 12-0-tet-radecanoylphorbol-13-acetate. Relationship with an inhibitor of plasminogen activator. J. Biol. Chem. 262: 2283–2290.

    PubMed  CAS  Google Scholar 

  39. Westerhausen DR, Hopkins WE, Billadello JJ. (1991) Multiple transforming growth factor-β-inducible elements regulate expression of the plasminogen activator inhibitor type-1 gene in Hep G2 cells. J. Biol. Chem. 266: 1092–1100.

    PubMed  CAS  Google Scholar 

  40. Fujii S, Sobel BE. (1990) Induction of plasminogen activator inhibitor by products released from platelets. Circulation 82: 1485–1493.

    Article  CAS  PubMed  Google Scholar 

  41. Herberg L, Coleman DL. (1977) Laboratory animals exhibiting obesity and diabetes syndromes. Metabolism 26: 59–99.

    Article  CAS  PubMed  Google Scholar 

  42. Pekary AE, Berg L, Wang J, Lee P, Dubinett SM, Hershman JM. (1995) TNF-alpha, TSH, and aging regulate TGF-beta synthesis and secretion in FRTL-5 rat thyroid cells. Am. J. Physiol. 268: R808–R815.

    PubMed  CAS  Google Scholar 

  43. Moses HL, Yang EY, Pietenpol JA. (1990) TGF-β stimulation and inhibition of cell proliferation: New mechanistic insights. Cell 63: 245–247.

    Article  CAS  PubMed  Google Scholar 

  44. Moses HL, Coffey Jr RJ, Leof EB, Lyons RM, Keski-Oja J. (1987) Transforming growth factor beta regulation of cell proliferation. J. Cell Physiol. 5: 1–7.

    Article  Google Scholar 

  45. Bortell R, van Wijnen AJ, Ramsey-Ewing AL, Stein GS, Stein JL. (1992) Differential regulation of H4 histone gene expression in 3T3-L1 pre-adipocytes during arrest of proliferation following contact inhibition or differentiation and its modulation by TGF beta 1. J. Cell. Biochem. 50: 62–72.

    Article  CAS  PubMed  Google Scholar 

  46. Jeoung DI, Tang B, Sonenberg M. (1995) Mitogenic response to TGF-beta in 3T3-F442A cells. Biochem. Biophys. Res. Commun. 216: 964–969.

    Article  CAS  PubMed  Google Scholar 

  47. Butterwith SC, Goddard C. (1991) Regulation of DNA synthesis in chicken adipocyte precursor cells by insulin-like growth factors, platelet-derived growth factor and transforming growth factor-beta. J. Endocrinol. 131: 203–209.

    Article  CAS  PubMed  Google Scholar 

  48. Vassaux G, Negrel R, Ailhaud G, Gaillard D. (1994) Proliferation and differentiation of rat adipose precursor cells in chemically defined medium: Differential action of anti-adipogenic agents. J. Cell. Physiol. 161: 249–256.

    Article  CAS  PubMed  Google Scholar 

  49. Pfeiffer A, Schatz H. (1995) Diabetic microvascular complications and growth factors. Exp. Clin. Endocrinol. Diabetes 103: 7–14.

    Article  CAS  PubMed  Google Scholar 

  50. Border WA. (1994) Transforming growth factor-beta and the pathogenesis of glomerular diseases. Curr. Opin. Nephrol. Hypertension 3: 54–58.

    Article  CAS  Google Scholar 

  51. Arrants J. (1994) Hyperinsulinemia and cardiovascular risk. Heart Lung 23: 118–122.

    PubMed  CAS  Google Scholar 

  52. Sowers JR. (1992) Insulin resistance, hyperinsulinemia, dyslipidemia, hypertension, and accelerated atherosclerosis. J. Clin. Pharmacol. 32: 529–535.

    Article  CAS  PubMed  Google Scholar 

  53. Sharma K, Ziyadeh FN. (1995) Hyperglycemia and diabetic kidney disease. The case for transforming growth factor-beta as a key mediator. Diabetes 44: 1139–1146.

    Article  CAS  PubMed  Google Scholar 

  54. Lundgren CH, Brown SL, Nordt TK, Sobel BE, Fujii S. (1996) Elaboration of type-1 plasminogen activator inhibitor from adipocytes: A potential pathogenetic link between obesity and cardiovascular disease. Circulation 93: 106–110.

    Article  CAS  PubMed  Google Scholar 

  55. Remade C, and Gregoire F. (1992) Cellular and molecular biology in the study of the physiopathology of obesity. Acta Clinica Belgica 14: (Suppl.) 3–12.

    Article  Google Scholar 

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Acknowledgments

This work was supported by grant HL 47819 to DJL. The authors thank T. Thinnes for her technical assistance. In addition we thank M. McRae for her expert secretarial skills.

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  1. Department of Vascular Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, VB-3, La Jolla, California, 92037, USA

    Fahumiya Samad, Koji Yamamoto, Manjula Pandey & David J. Loskutoff Ph.D.

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  1. Fahumiya Samad
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  2. Koji Yamamoto
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Samad, F., Yamamoto, K., Pandey, M. et al. Elevated Expression of Transforming Growth Factor-β in Adipose Tissue from Obese Mice. Mol Med 3, 37–48 (1997). https://doi.org/10.1007/BF03401666

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