Adipose Tissue Expression of Gelatinases in Mouse Models of Obesity

Summary

Following the observation by Brown et al. (Am J Physiol 1997; 272: C937-49) that primary rat adipocytes in culture secrete gelatinase A (MMP-2), we have evaluated gelatinase expression in adipose tissue with the use of mouse models of obesity. Wild-type mice were kept on a standard fat diet (SFD) or on a high fat diet (42% fat, HFD) and genetically obese db/db mice were kept on SFD; gonadal and subcutaneous fat pads were removed and analysed ex vivo. These studies revealed that: 1) the HFD induced adipocyte hypertrophy; 2) after 32 weeks, significantly higher levels of 70 kDa (p <0.05) and 65 kDa proMMP-2 (p < 0.01) were observed in extracts of gonadal fat pads of mice on HFD; 3) the contribution of active MMP-2 to the total level was comparable in SFD and HFD groups (20 to 30%); and 4) gelatinase B (MMP-9) was not consistently detected. These findings were confirmed by gelatin zymography and by mRNA determination using competitive RT-PCR. The presence of MMP-2 in the adipose tissue was confirmed immunologically and its localization in adipocytes revealed by immunogold electron microscopy. The potential functional role of MMP-2 in adipose tissue remains to be determined.

Key words

Obesity - matrix metalloproteinases - gelatinase

• References

• 1 Crandall DL, Hausman GJ, Kral JG. A review of the microcirculation of adipose tissue: anatomic, metabolic and angiogenic perspectives. Microcirculation 1997; 4: 211-32.
• 2 Dollery CM, McEwan JR, Henney AM. Matrix metalloproteinases and cardiovascular disease. Circ Res 1995; 77: 863-8.
• 3 Carmeliet P, Collen D. Development and disease in proteinase-deficient mice: role of the plasminogen, matrix metalloproteinase and coagulation system. Thromb Res 1998; 91: 255-85.
• 4 Samad F, Yamamoto K, Loskutoff DJ. 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 1996; 97: 37-46.
• 5 Alessi MC, Peiretti F, Morange P, Henry M, Nalbone G, Juhan-Vague I. Production of plasminogen activator inhibitor 1 by human adipose tissue. Possible link between viscaral fat accumulation and vascular disease. Diabetes 1997; 46: 860-7.
• 6 Juhan-Vague I, Alessi MC. Regulation of fibrinolysis in the development of atherothrombosis: role of adipose tissue. Thromb Haemost 1999; 82: 832-6.
• 7 Samad F, Loskutoff DJ. Hemostatic gene expression and vascular disease in obesity: insights from studies of genetically obese mice. Thromb Haemost 1999; 82: 742-7.
• 8 Morange PE, Lijnen HR, Alessi MC, Kopp F, Collen D, Juhan-Vague I. Influence of PAI-1 on adipose tissue growth and on metabolic parameters in a murine model of diet-induced obesity. Arterioscler Thromb Vasc Biol 2000; 20: 1150-4.
• 9 Lijnen HR, Collen D. Matrix metalloproteinase system deficiencies and matrix degradation. Thromb Haemost 1999; 82: 837-45.
• 10 Brown LM, Fox HL, Hazen SA, LaNoue KF, Rannels SR, Lynch CJ. Role of the matrixin MMP-2 in multicellular organization of adipocytes cultured in basement membrane components. Am J Physiol 1997; 272: C937-C49.
• 11 Sjöström L, Bjöntorp P, Vrana J. Microscopic fat cell size measurements on frozen-cut adipose tissue in comparison with automatic determinations of osmium-fixed fat cells. J Lipid Res 1971; 12: 521-30.
• 12 Folch J, Lees M, Sloane Stanley GH. A simple method for the isolation and purification of total lipids from animal tissue. J Biol Chem 1957; 232: 497-508.
• 13 Gliemann J, Osterlind K, Vinten J, Gammeltoft S. A procedure for measurements of distribution spaces in isolated fat cells. Biochim Biophys Acta 1972; 286: 1-9.
• 14 Kleiner DE, Stetler Stevenson WG. Quantitative zymography: detection of picogram quantities of gelatinases. Anal Biochem 1994; 218: 325-9.
• 15 Lijnen HR, Van Hoef B, Lupu F, Moons L, Carmeliet P, Collen D. Function of plasminogen/plasmin and matrix metalloproteinase systems after vascular injury in mice with targeted inactivation of fibrinolytic system genes. Arterioscler Thromb Vasc Biol 1998; 18: 1035-45.
• 16 Lijnen HR, Silence J, Lemmens G, Frederix L, Collen D. Regulation of gelatinase activity in mice with targeted inactivation of components of the plasminogen/plasmin system. Thromb Haemost 1998; 79: 1171-6.
• 17 Nie GY, Wang J, Li Y, Salamonsen LA. Construction and application of a multispecific competitor to quantify mRNA of matrix metalloproteinases and their tissue inhibitors in small human biopsies. J Biochem Biophys Methods 1999; 40: 81-99.
• 18 Lijnen HR, Lupu F, Moons L, Carmeliet P, Goulding D, Collen D. Temporal and topographic matrix metalloproteinase expression after vascular injury in mice. Thromb Haemost 1999; 81: 799-807.
• 19 Declerck PJ, Verstreken M, Collen D. Immunoassay of murine t-PA, u-PA and PAI-1 using monoclonal antibodies raised in gene-inactivated mice. Thromb Haemost 1995; 74: 1305-9.
• 20 Flier JS. The adipocyte: storage depot or node on the energy information superhighway?. Cell 1995; 80: 15-8.
• 21 Silverman KJ, Lund DP, Zetter BR, Lainey LL, Shahood JA, Freiman DG, Folkman J, Barger AC. Angiogenic activity of adipose tissue. Biochem Biophys Res Commun 1988; 153: 347-52.
• 22 Claffey KP, Wilkison WO, Spiegelman BM. Vascular endothelial growth factor; Regulation by cell differentiation and activated second messenger pathways. J Biol Chem 1992; 267: 16317-22.
• 23 Bouloumie A, Lafontan M, Busse R. Leptin, the product of Ob gene, promotes angiogenesis. Circ Res 1998; 83: 1059-66.
• 24 Darimont C, Saint-Marc P, Ailhaud G, Negrel R. Modulation of vascular tone and glycerol levels measured by in situ microdialysis in rat adipose tissue. Am J Physiol 1996; 271: E631-5.
• 25 Levine JA, Jensen MD, Eberhardt NL, O’Brien T. Adipocyte macrophage colony stimulating factor is a mediator of adipose tissue growth. J Clin Invest 1998; 101: 1557-64.
• 26 Negrel R, Gaillard D, Ailhaud G. Prostacyclin as a potent effector of adipose cell differentiation. Biochem J 1989; 257: 399-405.
• 27 Nougues J, Reyne Y, Barenton B, Chery T, Garandel V, Soriano J. Differentiation of adipocyte precursors in a serum-free medium is influenced by glucocorticoids and endogenously produced insulin-like growth factor-I. Int J, Obes 1993; 17: 159-67.
• 28 Rahimi N, Tremblay E, MacAdam L, Roberts A, Elliot B. Autocrine secretion of TGF-beta1 and TGF-beta2 by preadipocytes and adipocytes: a potent negative regulator of adipocyte differentiation and proliferation of mammary carcinoma cells. In Vitro Cell Dev Biol Anim 1998; 34: 412-20.
• 29 Nakajima I, Yamaguchi T, Ozutsumi K, Aso H. Adipose tissue extracellular matrix: newly organized by adipocytes during differentiation. Differentiation 1998; 63: 193-200.
• 30 Itoh T, Ikeda T, Gomi H, Nakao S, Suzuki T, Itohara S. Unaltered secretion of β-amyloid precursor protein in gelatinase A (matrix metalloproteinase 2)-deficient mice. J Biol Chem 1997; 272: 22389-92.
• 31 Itoh T, Tanioka M, Yoshida H, Yoshioka T, Nishimoto H, Itohara S. Reduced angiogenesis and tumor progression in gelatinase A-deficient mice. Cancer Res 1998; 58: 1048-51.
• 32 Digirolamo M, Fine JB, Tagra K, Rossmanith R. Qualitative regional differences in adipose tissue growth and cellularity in male Wistar rats fed ad libitum. Am J Physiol 1998; 274: R1460-7.
• 33 Spiegelman BM, Flier JS. Adipogenesis and obesity: rounding out the big picture. Cell 1996; 87: 377-89.
• 34 Geloen A, Roy PE, Bukowecki LJ. Regression of white adipose tissue in diabetic rats. Am J Physiol 1989; 257: E547-E53.