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Journal of Molecular Medicine

, Volume 87, Issue 5, pp 481–492 | Cite as

NF-κB blockade upregulates Bax, TSP-1, and TSP-2 expression in rat granulation tissue

  • Daniela De Stefano
  • Giancarlo Nicolaus
  • Maria Chiara Maiuri
  • Daniela Cipolletta
  • Lorenzo Galluzzi
  • Maria Pia Cinelli
  • Gianfranco Tajana
  • Teresa Iuvone
  • Rosa CarnuccioEmail author
Original Article

Abstract

Several diseases are characterized by chronic inflammation, a condition frequently associated with angiogenesis and fibrogenesis that account for the development of granulation tissue. Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-κB) is a crucial modulator of intracellular prosurvival signaling pathways and is implicated in the pathogenesis of inflammatory process. In this study, we have investigated the role of NF-κB in the angiogenic and fibrogenic response induced by λ-carrageenin in a rat model of chronic inflammation at 1, 3, and 5 days. The subcutaneous implant of λ-carrageenin-soaked sponges in rat induced a time-related increase of granulation tissue formation accompanied by intense neovascularization. These λ-carrageenin-induced changes were significantly reduced by coinjection of wild-type oligodeoxynucleotide (WT ODN) decoy to NF-κB. Molecular, morphological, and ultrastructural analysis performed on whole granulation tissue demonstrated: (1) inhibition of NF-κB/DNA binding activity; (2) downregulation of cyclooxygenase-2, matrix metalloproteinase-9, tumor necrosis factor-α, and vascular endothelial growth factor; (3) upregulation of thrombospondin (TSP)-1 at 1 day and TSP-2 at 5 days; and (4) increase in Bax to Bcl-2 ratio. Our findings show that the blockade of NF-κB activation by WT ODN decoy prevents the development of granulation tissue induced by λ-carrageenin-soaked sponge implant upregulating Bax as well as TSP-1 and TSP-2 expression.

Keywords

Angiogenesis Apoptosis Bcl-2 protein family Granulation tissue NF-κB Phagocytosis 

Notes

Acknowledgements

Prof. M.L. Del Basso De Caro is gratefully thanked for the assistance and technical support. We also kindly acknowledge Dr. M.T. Ribecco for her constant help and invaluable suggestions. This work has been supported by the Italian Government.

Supplementary material

109_2009_443_MOESM1_ESM.doc (22 kb)
Supplementry legend information (DOC 22KB).
109_2009_443_MOESM2_ESM.ppt (1.7 mb)
Supplementry figures (PPT 1.71 MB).

References

  1. 1.
    Jackson JR, Bolognese B, Kircher CH, Marshall LA, Winkler JD (1997) Modulation of angiogenesis in a model of chronic inflammation. Inflamm Res 46(Suppl 2):S129–S130CrossRefPubMedGoogle Scholar
  2. 2.
    Majno G (1998) Chronic inflammation: links with angiogenesis and wound healing. Am J Pathol 153:1035–1039PubMedGoogle Scholar
  3. 3.
    Baud V, Karin M (2001) Signal transduction by tumor necrosis factor and its relatives. Trends Cell Biol 11:372–377CrossRefPubMedGoogle Scholar
  4. 4.
    Cross MJ, Claesson-Welsh L (2001) FGF and VEGF function in angiogenesis: signalling pathways, biological responses and therapeutic inhibition. Trends Pharmacol Sci 22:201–207CrossRefPubMedGoogle Scholar
  5. 5.
    Neufeld G, Cohen T, Gengrinovitch S, Poltorak Z (1999) Vascular endothelial growth factor (VEGF) and its receptors. FASEB J 13:9–22PubMedGoogle Scholar
  6. 6.
    Ghosh AK, Hirasawa N, Niki H, Ohuchi K (2000) Cyclooxygenase-2-mediated angiogenesis in carrageenin-induced granulation tissue in rats. J Pharmacol Exp Ther 295:802–809PubMedGoogle Scholar
  7. 7.
    Bornstein P, Armstrong LC, Hankenson KD, Kyriakides TR, Yang Z (2000) Thrombospondin 2, a matricellular protein with diverse functions. Matrix Biol 19:557–568CrossRefPubMedGoogle Scholar
  8. 8.
    Streit M, Velasco P, Riccardi L, Spencer L, Brown LF, Janes L, Lange-Asschenfeldt B, Yano K, Hawighorst T, Iruela-Arispe L, Detmar M (2000) Thrombospondin-1 suppresses wound healing and granulation tissue formation in the skin of transgenic mice. EMBO J 19:3272–3282CrossRefPubMedGoogle Scholar
  9. 9.
    Park YW, Kang YM, Butterfield J, Detmar M, Goronzy JJ, Weyand CM (2004) Thrombospondin 2 functions as an endogenous regulator of angiogenesis and inflammation in rheumatoid arthritis. Am J Pathol 165:2087–2098PubMedGoogle Scholar
  10. 10.
    Nör JE, Peters MC, Christensen JB, Sutorik MM, Linn S, Khan MK, Addison CL, Mooney DJ, Polverini PJ (2000) Thrombospondin-1 induces endothelial cell apoptosis and inhibits angiogenesis by activating the caspase death pathway. J Vasc Res 37:209–218CrossRefPubMedGoogle Scholar
  11. 11.
    Kroemer G, Galluzzi L, Brenner C (2007) Mitochondrial membrane permeabilization in cell death. Physiol Rev 87:99–163CrossRefPubMedGoogle Scholar
  12. 12.
    Tak PP, Firestein GS (2001) NF-kappaB: a key role in inflammatory diseases. J Clin Invest 107:7–11CrossRefPubMedGoogle Scholar
  13. 13.
    Farina AR, Tacconelli A, Vacca A, Maroder M, Gulino A, Mackay AR (1999) Transcriptional up-regulation of matrix metalloproteinase-9 expression during spontaneous epithelial to neuroblast phenotype conversion by SK-N-SH neuroblastoma cells, involved in enhanced invasivity, depends upon GT-box and nuclear factor kappaB elements. Cell Growth Differ 10:353–367PubMedGoogle Scholar
  14. 14.
    Sid B, Sartelet H, Bellon G, El Btaouri H, Rath G, Delorme N, Haye B, Martiny L (2004) Thrombospondin 1: a multifunctional protein implicated in the regulation of tumor growth. Crit Rev Oncol Hematol 49:245–258CrossRefPubMedGoogle Scholar
  15. 15.
    Yang YL, Chuang LY, Guh JY, Liu SF, Hung MY, Liao TN, Huang YL (2004) Thrombospondin-1 mediates distal tubule hypertrophy induced by glycated albumin. Biochem J 379:89–97CrossRefPubMedGoogle Scholar
  16. 16.
    Lin B, Williams-Skipp C, Tao Y, Schleicher MS, Cano LL, Duke RC, Scheinman RI (1999) NF-kappaB functions as both a proapoptotic and antiapoptotic regulatory factor within a single cell type. Cell Death Differ 6:570–582CrossRefPubMedGoogle Scholar
  17. 17.
    Radhakrishnan SK, Kamalakaran S (2006) Pro-apoptotic role of NF-kappaB: implications for cancer therapy. Biochim Biophys Acta 1766:53–62PubMedGoogle Scholar
  18. 18.
    Beg AA, Baltimore D (1996) An essential role for NF-kappaB in preventing TNF-alpha-induced cell death. Science 274:782–784CrossRefPubMedGoogle Scholar
  19. 19.
    Van Antwerp DJ, Martin SJ, Kafri T, Green DR, Verma IM (1996) Suppression of TNF-alpha-induced apoptosis by NF-kappaB. Science 274:787–789CrossRefPubMedGoogle Scholar
  20. 20.
    Wang CY, Mayo MW, Korneluk RG, Goeddel DV, Baldwin AS Jr (1998) NF-kappaB antiapoptosis: induction of TRAF1 and TRAF2 and c-IAP1 and c-IAP2 to suppress caspase-8 activation. Science 281:1680–1683CrossRefPubMedGoogle Scholar
  21. 21.
    De Smaele E, Zazzeroni F, Papa S, Nguyen DU, Jin R, Jones J, Cong R, Franzoso G (2001) Induction of gadd45beta by NF-kappaB downregulates pro-apoptotic JNK signalling. Nature 414:308–313CrossRefPubMedGoogle Scholar
  22. 22.
    Karin M, Lin A (2002) NF-kappaB at the crossroads of life and death. Nat Immunol 3:221–227CrossRefPubMedGoogle Scholar
  23. 23.
    D’Acquisto F, de Cristofaro F, Maiuri MC, Tajana G, Carnuccio R (2001) Protective role of nuclear factor kappa B against nitric oxide-induced apoptosis in J774 macrophages. Cell Death Differ 8:144–151CrossRefPubMedGoogle Scholar
  24. 24.
    Fujihara S, Ward C, Dransfield I, Hay RT, Uings IJ, Hayes B, Farrow SN, Haslett C, Rossi AG (2002) Inhibition of nuclear factor-kappaB activation un-masks the ability of TNF-alpha to induce human eosinophil apoptosis. Eur J Immunol 32:457–466CrossRefPubMedGoogle Scholar
  25. 25.
    Ward C, Chilvers ER, Lawson MF, Pryde JG, Fujihara S, Farrow SN, Haslett C, Rossi AG (1999) NF-kappaB activation is a critical regulator of human granulocyte apoptosis in vitro. J Biol Chem 274:4309–4318CrossRefPubMedGoogle Scholar
  26. 26.
    Wu M, Lee H, Bellas RE, Schauer SL, Arsura M, Katz D, FitzGerald MJ, Rothstein TL, Sherr DH, Sonenshein GE (1996) Inhibition of NF-kappaB/Rel induces apoptosis of murine B cells. EMBO J 15:4682–4690PubMedGoogle Scholar
  27. 27.
    Maiuri MC, Tajana G, Iuvone T, De Stefano D, Mele G, Ribecco MT, Cinelli MP, Romano MF, Turco MC, Carnuccio R (2004) Nuclear factor-kappaB regulates inflammatory cell apoptosis and phagocytosis in rat carrageenin-sponge implant model. Am J Pathol 165:115–126PubMedGoogle Scholar
  28. 28.
    Iuvone T, Carnuccio R, Di Rosa M (1994) Modulation of granuloma formation by endogenous nitric oxide. Eur J Pharmacol 265:89–92CrossRefPubMedGoogle Scholar
  29. 29.
    De Filippis D, Russo A, De Stefano D, Maiuri MC, Esposito G, Cinelli MP, Pietropaolo C, Carnuccio R, Russo G, Iuvone T (2007) Local administration of WIN 55,212-2 reduces chronic granuloma-associated angiogenesis in rat by inhibiting NF-kappaB activation. J Mol Med 85:635–645CrossRefPubMedGoogle Scholar
  30. 30.
    Muramatsu M, Katada J, Hayashi I, Majima M (2000) Chymase as a proangiogenic factor. A possible involvement of chymase-angiotensin-dependent pathway in the hamster sponge angiogenesis model. J Biol Chem 275:5545–5552CrossRefPubMedGoogle Scholar
  31. 31.
    Aggarwal BB, Shishodia S, Sandur SK, Pandey MK, Sethi G (2006) Inflammation and cancer: how hot is the link? Biochem Pharmacol 72:1605–1621CrossRefPubMedGoogle Scholar
  32. 32.
    Lin WW, Karin M (2007) A cytokine-mediated link between innate immunity, inflammation, and cancer. J Clin Invest 117:1175–1183CrossRefPubMedGoogle Scholar
  33. 33.
    Tan TT, Coussens LM (2007) Humoral immunity, inflammation and cancer. Curr Opin Immunol 19:209–216CrossRefPubMedGoogle Scholar
  34. 34.
    Meneghin A, Hogaboam CM (2007) Infectious disease, the innate immune response, and fibrosis. J Clin Invest 117:530–538CrossRefPubMedGoogle Scholar
  35. 35.
    Savill J (1997) Apoptosis in resolution of inflammation. J Leukoc Biol 61:375–380PubMedGoogle Scholar
  36. 36.
    Matsuyama W, Watanabe M, Shirahama Y, Mitsuyama H, Higashimoto I, Osame M, Arimura K (2006) Discoidin domain receptor 1 contributes to the survival of lung fibroblast in idiopathic pulmonary fibrosis. Am J Pathol 168:866–877CrossRefPubMedGoogle Scholar
  37. 37.
    Bentires-Alj M, Dejardin E, Viatour P, Van Lint C, Froesch B, Reed JC, Merville MP, Bours V (2001) Inhibition of the NF-kappa B transcription factor increases Bax expression in cancer cell lines. Oncogene 20:2805–2813CrossRefPubMedGoogle Scholar
  38. 38.
    Feuillard J, Schuhmacher M, Kohanna S, Asso-Bonnet M, Ledeur F, Joubert-Caron R, Bissieres P, Polack A, Bornkamm GW, Raphael M (2000) Inducible loss of NF-kappaB activity is associated with apoptosis and Bcl-2 down-regulation in Epstein-Barr virus-transformed B lymphocytes. Blood 95:2068–2075PubMedGoogle Scholar
  39. 39.
    Grossmann M, O’Reilly LA, Gugasyan R, Strasser A, Adams JM, Gerondakis S (2000) The anti-apoptotic activities of Rel and RelA required during B-cell maturation involve the regulation of Bcl-2 expression. EMBO J 19:6351–6360CrossRefPubMedGoogle Scholar
  40. 40.
    Dikshit P, Chatterjee M, Goswami A, Mishra A, Jana NR (2006) Aspirin induces apoptosis through the inhibition of proteasome function. J Biol Chem 281:29228–29235CrossRefPubMedGoogle Scholar
  41. 41.
    Agah A, Kyriakides TR, Letrondo N, Bjorkblom B, Bornstein P (2004) Thrombospondin 2 levels are increased in aged mice: consequences for cutaneous wound healing and angiogenesis. Matrix Biol 22:539–547CrossRefPubMedGoogle Scholar
  42. 42.
    Agah A, Kyriakides TR, Lawler J, Bornstein P (2002) The lack of thrombospondin-1 (TSP1) dictates the course of wound healing in double-TSP1/TSP2-null mice. Am J Pathol 161:831–839PubMedGoogle Scholar
  43. 43.
    Cinatl J Jr, Bittoova M, Margraf S, Vogel JU, Cinatl J, Preiser W, Doerr HW (2000) Cytomegalovirus infection decreases expression of thrombospondin-1 and -2 in cultured human retinal glial cells: effects of antiviral agents. J Infect Dis 182:643–651CrossRefPubMedGoogle Scholar
  44. 44.
    Kuprash DV, Udalova IA, Turetskaya RL, Rice NR, Nedospasov SA (1995) Conserved kappa B element located downstream of the tumor necrosis factor alpha gene: distinct NF-kappa B binding pattern and enhancer activity in LPS activated murine macrophages. Oncogene 11:97–106PubMedGoogle Scholar
  45. 45.
    Schmedtje JF, Ji YS, Liu WL, DuBois RN, Runge MS (1997) Hypoxia induces cyclooxygenase-2 via the NF-kappaB p65 transcription factor in human vascular endothelial cells. J Biol Chem 272:601–608CrossRefPubMedGoogle Scholar
  46. 46.
    Huang WC, Chan ST, Yang TL, Tzeng CC, Chen CC (2004) Inhibition of ICAM-1 gene expression, monocyte adhesion and cancer cell invasion by targeting IKK complex: molecular and functional study of novel alpha-methylene-gamma-butyrolactone derivatives. Carcinogenesis 25:1925–1934CrossRefPubMedGoogle Scholar
  47. 47.
    Ondrey FG, Dong G, Sunwoo J, Chen Z, Wolf JS, Crowl-Bancroft CV, Mukaida N, Van Waes C (1999) Constitutive activation of transcription factors NF-(kappa)B, AP-1, and NF-IL6 in human head and neck squamous cell carcinoma cell lines that express pro-inflammatory and pro-angiogenic cytokines. Mol Carcinog 26:119–129CrossRefPubMedGoogle Scholar
  48. 48.
    Rodriguez-Manzaneque JC, Lane TF, Ortega MA, Hynes RO, Lawler J, Iruela-Arispe ML (2001) Thrombospondin-1 suppresses spontaneous tumor growth and inhibits activation of matrix metalloproteinase-9 and mobilization of vascular endothelial growth factor. Proc Natl Acad Sci U S A 98:12485–12490CrossRefPubMedGoogle Scholar
  49. 49.
    Sennlaub F, Valamanesh F, Vazquez-Tello A, El-Asrar AM, Checchin D, Brault S, Gobeil F, Beauchamp MH, Mwaikambo B, Courtois Y, Geboes K, Varma DR, Lachapelle P, Ong H, Behar-Cohen F, Chemtob S (2003) Cyclooxygenase-2 in human and experimental ischemic proliferative retinopathy. Circulation 108:198–204CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Daniela De Stefano
    • 1
  • Giancarlo Nicolaus
    • 2
  • Maria Chiara Maiuri
    • 1
  • Daniela Cipolletta
    • 2
  • Lorenzo Galluzzi
    • 3
  • Maria Pia Cinelli
    • 4
  • Gianfranco Tajana
    • 5
  • Teresa Iuvone
    • 6
  • Rosa Carnuccio
    • 1
    Email author
  1. 1.Dipartimento di Farmacologia Sperimentale, Facoltà di Scienze BiotecnologicheUniversità degli Studi di Napoli Federico IINaplesItaly
  2. 2.Istituto di Ricerche di Biologia Molecolare P. AngelettiPomeziaItaly
  3. 3.INSERM U848, Institut Gustave RoussyVillejuifFrance
  4. 4.Dipartimento di Scienze Biomorfologiche e Funzionali, Facoltà di Medicina e ChirurgiaUniversità degli Studi di Napoli Federico IINaplesItaly
  5. 5.Dipartimento di Scienze Farmaceutiche, Facoltà di Medicina e ChirurgiaUniversità degli Studi di SalernoFiscianoItaly
  6. 6.Dipartimento di Farmacologia Sperimentale, Facoltà di FarmaciaUniversità degli Studi di Napoli Federico IINaplesItaly

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