Pflügers Archiv - European Journal of Physiology

, Volume 458, Issue 6, pp 1103–1114 | Cite as

Microarray analysis identifies matrix metalloproteinases (MMPs) as key genes whose expression is up-regulated in human adipocytes by macrophage-conditioned medium

  • Adrian O’Hara
  • Fei-Ling Lim
  • Dawn J. Mazzatti
  • Paul Trayhurn
Molecular and Genomic Physiology


White adipose tissue exhibits inflammation as tissue mass expands in obesity, involving macrophage infiltration and a direct inflammatory response by adipocytes. DNA microarrays and conditioned medium have been used to examine the effects of macrophages on global gene expression in human adipocytes. SGBS adipocytes, differentiated in culture, were treated with macrophage-conditioned medium (U937 cells) for 4 or 24 h; control cells received unconditioned medium. Agilent arrays comprising 44,000 probes were used to analyse gene expression. Microarray analysis identified 1,088 genes differentially expressed in response to the conditioned medium at both 4 and 24 h (754 up-regulated, 334 down-regulated at 24 h); these included genes associated with inflammation and macrophage infiltration. A cluster of matrix metalloproteinase genes were highly up-regulated at both time-points, including MMP1, MMP3, MMP9, MMP10, MMP12 and MMP19. At 4 and 24 h, MMP1 was the most highly up-regulated gene (>2,400-fold increase in mRNA at 24 h). ELISA measurements indicated that substantial quantities of MMP1 and MMP3 were released from adipocytes incubated with conditioned medium, with little release by control adipocytes. Treatment with TNFα induced substantial increases in MMP1 (>100-fold) and MMP3 (27-fold) mRNA level and MMP1 and MMP3 release in adipocytes, suggesting that this cytokine could contribute to the stimulation of MMP expression by macrophages. In conclusion, macrophage-secreted factors induce a major inflammatory response in human adipocytes, with expression of MMP family members being strongly up-regulated. The induction of MMP1 and other MMPs suggests that macrophages stimulate tissue remodelling during adipose tissue expansion in obesity.


Adipose tissue Inflammation Macrophages Obesity TNFα 





macrophage conditioned


matrix metalloproteinase


tumour necrosis factor-α





We are grateful to Dr. Stuart Wood and to Mr. Leif Hunter for their help and advice. PT is a member of COST BM0602. AOH is in receipt of an Industrial CASE Studentship from the Biotechnology and Biological Sciences Research Council (UK), which is also thanked for grant support to PT.


The authors have no conflicts of interests to declare; this study has no commercial implications.

Supplementary material

424_2009_693_MOESM1_ESM.doc (836 kb)


  1. 1.
    Ahima RS, Flier JS (2000) Adipose tissue as an endocrine organ. Trends Endocrinol Metab 11:327–332PubMedCrossRefGoogle Scholar
  2. 2.
    Alvarez-Llamas G, Szalowska E, de Vries MP, Weening D, Landman K, Hoek A, Wolffenbuttel BH, Roelofsen H, Vonk RJ (2007) Characterization of the human visceral adipose tissue secretome. Mol Cell Proteomics 6:589–600PubMedCrossRefGoogle Scholar
  3. 3.
    Aoki N, Jin-no S, Nakagawa Y, Asai N, Arakawa E, Tamura N, Tamura T, Matsuda T (2007) Identification and characterization of microvesicles secreted by 3T3–L1 adipocytes: redox- and hormone-dependent induction of milk fat globule-epidermal growth factor 8-associated microvesicles. Endocrinology 148:3850–3862PubMedCrossRefGoogle Scholar
  4. 4.
    Bulló M, Garcia-Lorda P, Megias I, Salas-Salvado J (2003) Systemic inflammation, adipose tissue tumor necrosis factor, and leptin expression. Obes Res 11:525–531PubMedCrossRefGoogle Scholar
  5. 5.
    Cancello R, Tordjman J, Poitou C, Guilhem G, Bouillot JL, Hugol D, Coussieu C, Basdevant A, Hen AB, Bedossa P, Guerre-Millo M, Clément K (2006) Increased infiltration of macrophages in omental adipose tissue is associated with marked hepatic lesions in morbid human obesity. Diabetes 55:1554–1561PubMedCrossRefGoogle Scholar
  6. 6.
    Chavey C, Mari B, Monthouel MN, Bonnafous S, Anglard P, Van Obberghen E, Tartare-Deckert S (2003) Matrix metalloproteinases are differentially expressed in adipose tissue during obesity and modulate adipocyte differentiation. J Biol Chem 278:11888–11896PubMedCrossRefGoogle Scholar
  7. 7.
    Chizzolini C, Rezzonico R, De Luca C, Burger D, Dayer JM (2000) Th2 cell membrane factors in association with IL-4 enhance matrix metalloproteinase-1 (MMP-1) while decreasing MMP-9 production by granulocyte-macrophage colony-stimulating factor-differentiated human monocytes. J Immunol 164:5952–5960PubMedGoogle Scholar
  8. 8.
    Cinti S, Mitchell G, Barbatelli G, Murano I, Ceresi E, Faloia E, Wang S, Fortier M, Greenberg AS, Obin MS (2005) Adipocyte death defines macrophage localization and function in adipose tissue of obese mice and humans. J Lipid Res 46:2347–2355PubMedCrossRefGoogle Scholar
  9. 9.
    Constant V, Gagnon A, Landry A, Sorisky A (2006) Macrophage-conditioned medium inhibits the differentiation of 3T3–L1 and human abdominal preadipocytes. Diabetologia 49:1402–1411PubMedCrossRefGoogle Scholar
  10. 10.
    Croissandeau G, Chrétien M, Mbikay M (2002) Involvement of matrix metalloproteinases in the adipose conversion of 3T3–L1 preadipocytes. Biochem J 364:739–746PubMedCrossRefGoogle Scholar
  11. 11.
    Curat C, Wegner V, Sengenѐs C, Miranville A, Tonus C, Busse R, Bouloumié A (2006) Macrophages in human visceral adipose tissue: increased accumulation in obesity and a source of resistin and visfatin. Diabetologia 49:744–747PubMedCrossRefGoogle Scholar
  12. 12.
    Derosa G, Ferrari I, D'Angelo A, Tinelli C, Salvadeo SA, Ciccarelli L, Piccinni MN, Gravina A, Ramondetti F, Maffioli P, Cicero AF (2008) Matrix metalloproteinase-2 and -9 levels in obese patients. Endothelium 15:219–224PubMedCrossRefGoogle Scholar
  13. 13.
    Do MS, Nam SY, Hong SE, Kim KW, Duncan JS, Beattie JH, Trayhurn P (2002) Metallothionein gene expression in human adipose tissue from lean and obese subjects. Horm Metab Res 34:348–351PubMedCrossRefGoogle Scholar
  14. 14.
    Engström G, Hedblad B, Stavenow L, Lind P, Janzon L, Lindgarde F (2003) Inflammation-sensitive plasma proteins are associated with future weight gain. Diabetes 52:2097–2101PubMedCrossRefGoogle Scholar
  15. 15.
    Festa A, D'Agostino R, Williams K, Karter AJ, Mayer-Davis EJ, Tracy RP, Haffner SM (2001) The relation of body fat mass and distribution to markers of chronic inflammation. Int J Obes 25:1407–1415CrossRefGoogle Scholar
  16. 16.
    Guerre-Millo M (2002) Adipose tissue hormones. J Endocrinol Investig 25:855–861Google Scholar
  17. 17.
    Hausman GJ, Poulos SP, Richardson RL, Barb CR, Andacht T, Kirk HC, Mynatt RL (2006) Secreted proteins and genes in fetal and neonatal pig adipose tissue and stromal-vascular cells. J Anim Sci 84:1666–1681PubMedCrossRefGoogle Scholar
  18. 18.
    Hirasaka K, Kohno S, Goto J, Furochi H, Mawatari K, Harada N, Hosaka T, Nakaya Y, Ishidoh K, Obata T, Ebina Y, Gu H, Takeda S, Kishi K, Nikawa T (2007) Deficiency of CBL-B gene enhances infiltration and activation of macrophages in adipose tissue and causes peripheral insulin resistance in mice. Diabetes 56:2511–2522PubMedCrossRefGoogle Scholar
  19. 19.
    Hotamisligil GS (2006) Inflammation and metabolic disorders. Nature 444:860–867PubMedCrossRefGoogle Scholar
  20. 20.
    Hotamisligil GS, Shargill NS, Spiegelman BM (1993) Adipose expression of tumor-necrosis-factor-α—direct role in obesity-linked insulin resistance. Science 259:87–91PubMedCrossRefGoogle Scholar
  21. 21.
    Karmakar S, Das C (2002) Regulation of trophoblast invasion by IL-1β and TGF-β1. Am J Reprod Immunol 48:210–219PubMedCrossRefGoogle Scholar
  22. 22.
    Kawamura T, Murakami K, Bujo H, Unoki H, Jiang M, Nakayama T, Saito Y (2008) Matrix metalloproteinase-3 enhances the free fatty acids-induced VEGF expression in adipocytes through toll-like receptor 2. Exp Biol Med 233:1213–1221CrossRefGoogle Scholar
  23. 23.
    Klatt AR, Klinger G, Neumuller O, Eidenmuller B, Wagner I, Achenbach T, Aigner T, Bartnik E (2006) TAK1 downregulation reduces IL-1β induced expression of MMP13, MMP1 and TNF-α. Biomed Pharmacother 60:55–61PubMedCrossRefGoogle Scholar
  24. 24.
    Lacasa D, Taleb S, Keophiphath M, Miranville A, Clément K (2007) Macrophage-secreted factors impair human adipogenesis: involvement of proinflammatory state in preadipocytes. Endocrinology 148:868–877PubMedCrossRefGoogle Scholar
  25. 25.
    Lumeng CN, Deyoung SM, Saltiel AR (2007) Macrophages block insulin action in adipocytes by altering expression of signaling and glucose transport proteins. Am J Physiol Endocrinol Metabol 292:E166–E174CrossRefGoogle Scholar
  26. 26.
    Maquoi E, Munaut C, Colige A, Collen D, Lijnen HR (2002) Modulation of adipose tissue expression of murine matrix metalloproteinases and their tissue inhibitors with obesity. Diabetes 51:1093–1101PubMedCrossRefGoogle Scholar
  27. 27.
    Mazzatti DJ, Smith MA, Oita RC, Lim FL, White AJ, Reid MB (2008) Muscle unloading-induced metabolic remodeling is associated with acute alterations in PPARδ and UCP-3 expression. Physiol Genomics 34:149–161PubMedCrossRefGoogle Scholar
  28. 28.
    Page-McCaw A, Ewald AJ, Werb Z (2007) Matrix metalloproteinases and the regulation of tissue remodelling. Nature Rev Mol Cell Biol 8:221–233CrossRefGoogle Scholar
  29. 29.
    Permana PA, Menge C, Reaven PD (2006) Macrophage-secreted factors induce adipocyte inflammation and insulin resistance. Biochem Biophys Res Commun 341:507–514PubMedCrossRefGoogle Scholar
  30. 30.
    Rajala MW, Scherer PE (2003) The adipocyte—at the crossroads of energy homeostasis, inflammation, and atherosclerosis. Endocrinology 144:3765–3773PubMedCrossRefGoogle Scholar
  31. 31.
    Rennie KL, Jebb SA (2005) Prevalence of obesity in Great Britain. Obes Rev 6:11–12PubMedCrossRefGoogle Scholar
  32. 32.
    Rifas L, Fausto A, Scott MJ, Avioli LV, Welgus HG (1994) Expression of metalloproteinases and tissue inhibitors of metalloproteinases in human osteoblast-like cells: differentiation is associated with repression of metalloproteinase biosynthesis. Endocrinology 134:213–221PubMedCrossRefGoogle Scholar
  33. 33.
    Rosen ED, Spiegelman BM (2006) Adipocytes as regulators of energy balance and glucose homeostasis. Nature 444:847–853PubMedCrossRefGoogle Scholar
  34. 34.
    Sampieri CL, Nuttall RK, Young DA, Goldspink D, Clark IM, Edwards DR (2008) Activation of p38 and JNK MAPK pathways abrogates requirement for new protein synthesis for phorbol ester mediated induction of select MMP and TIMP genes. Matrix Biol 27:128–138PubMedCrossRefGoogle Scholar
  35. 35.
    Suganami T, Nishida J, Ogawa Y (2005) A paracrine loop between adipocytes and macrophages aggravates inflammatory changes—role of free fatty acids and tumor necrosis factor α. Arterioscler Thromb Vasc Biol 25:2062–2068PubMedCrossRefGoogle Scholar
  36. 36.
    Sundström C, Nilsson K (1976) Establishment and characterization of a human histiocytic lymphoma cell line (U-937). Int J Cancer 17:565–577PubMedCrossRefGoogle Scholar
  37. 37.
    Trayhurn P, Beattie JH (2001) Physiological role of adipose tissue: white adipose tissue as an endocrine and secretory organ. Proc Nutr Soc 60:329–339PubMedCrossRefGoogle Scholar
  38. 38.
    Trayhurn P, Duncan JS, Wood AM, Beattie JH (2000) Metallothionein gene expression and secretion in white adipose tissue. Am J Physiol Regul Integr Comp Physiol 279:R2329–R2335PubMedGoogle Scholar
  39. 39.
    Trayhurn P, Duncan JS, Wood AM, Beattie JH (2000) Regulation of metallothionein gene expression and secretion in rat adipocytes differentiated from preadipocytes in primary culture. Horm Metab Res 32:542–547PubMedCrossRefGoogle Scholar
  40. 40.
    Trayhurn P, Wood IS (2004) Adipokines: inflammation and the pleiotropic role of white adipose tissue. Br J Nutr 92:347–355PubMedCrossRefGoogle Scholar
  41. 41.
    Trayhurn P, Wood IS (2005) Signalling role of adipose tissue: adipokines and inflammation in obesity. Biochem Soc Trans 33:1078–1081PubMedCrossRefGoogle Scholar
  42. 42.
    van Beek EA, Bakker AH, Kruyt PM, Vink C, Saris WH, Franssen-van Hal NLW, Keijer J (2008) Comparative expression analysis of isolated human adipocytes and the human adipose cell lines LiSa-2 and PAZ6. Int J Obes 32:912–921CrossRefGoogle Scholar
  43. 43.
    Wabitsch M, Brenner RE, Melzner I, Braun M, Moller P, Heinze E, Debatin KM, Hauner H (2001) Characterization of a human preadipocyte cell strain with high capacity for adipose differentiation. Int J Obes 25:8–15CrossRefGoogle Scholar
  44. 44.
    Wang B, Jenkins JR, Trayhurn P (2005) Expression and secretion of inflammation-related adipokines by human adipocytes differentiated in culture: integrated response to TNF-α. Am J Physiol Endocrinol Metab 288:E731–E740PubMedCrossRefGoogle Scholar
  45. 45.
    Wang B, Trayhurn P (2006) Acute and prolonged effects of TNF-alpha on the expression and secretion of inflammation-related adipokines by human adipocytes differentiated in culture. Pflügers Arch Eur J Physiol 452:418–427CrossRefGoogle Scholar
  46. 46.
    Wang B, Wood IS, Trayhurn P (2008) PCR arrays identify metallothionein-3 as a highly hypoxia-inducible gene in human adipocytes. Biochem Biophys Res Commun 368:88–93PubMedCrossRefGoogle Scholar
  47. 47.
    Watari M, Watari H, DiSanto ME, Chacko S, Shi GP, Strauss JF 3rd (1999) Pro-inflammatory cytokines induce expression of matrix-metabolizing enzymes in human cervical smooth muscle cells. Am J Pathol 154:1755–1762PubMedGoogle Scholar
  48. 48.
    Weisberg SP, McCann D, Desai M, Rosenbaum M, Leibel RL, Ferrante AW Jr (2003) Obesity is associated with macrophage accumulation in adipose tissue. J Clin Invest 112:1796–1808PubMedGoogle Scholar
  49. 49.
    Xu H, Barnes GT, Yang Q, Tan G, Yang D, Chou CJ, Sole J, Nichols A, Ross JS, Tartaglia LA, Chen H (2003) Chronic inflammation in fat plays a crucial role in the development of obesity-related insulin resistance. J Clin Invest 112:1821–1830PubMedGoogle Scholar
  50. 50.
    Yamashita A, Soga Y, Iwamoto Y, Asano T, Li Y, Abiko Y, Nishimura F (2008) DNA microarray analyses of genes expressed differentially in 3T3–L1 adipocytes co-cultured with murine macrophage cell line RAW264.7 in the presence of the toll-like receptor 4 ligand bacterial endotoxin. Int J Obesity 32:1725–1729 2005CrossRefGoogle Scholar
  51. 51.
    Yudkin JS, Stehouwer CDA, Emeis JJ, Coppack SW (1999) C-reactive protein in wealthy subjects: associations with obesity, insulin resistance, and endothelial dysfunction—a potential role for cytokines originating from adipose tissue? Arterioscler Thromb Vasc Biol 19:972–978PubMedGoogle Scholar
  52. 52.
    Zhang YY, Proenca R, Maffei M, Barone M, Leopold L, Friedman JM (1994) Positional cloning of the mouse obese gene and its human homolog. Nature 372:425–432PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Adrian O’Hara
    • 1
  • Fei-Ling Lim
    • 2
  • Dawn J. Mazzatti
    • 2
  • Paul Trayhurn
    • 1
  1. 1.Obesity Biology Research Unit, School of Clinical SciencesUniversity of LiverpoolLiverpoolUK
  2. 2.Unilever R&D DiscoverBedfordshireUK

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