Download PDF
Thromb Haemost 2004; 92(04): 858-866
DOI: 10.1160/TH04-04-0261
Endothelium and Vascular Development
Schattauer GmbH

Fibrinogen regulates the expression of inflammatory chemokines through NF-κB activation of endothelial cells

Min Guo
1   Hematology-Oncology Unit, Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York, USA
,
Sanjeev K. Sahni
1   Hematology-Oncology Unit, Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York, USA
,
Abha Sahni
1   Hematology-Oncology Unit, Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York, USA
,
Charles W. Francis
1   Hematology-Oncology Unit, Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York, USA
› Author AffiliationsFinancial support: This work was supported in part by Grant No. HL-30616 from the National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland.
Further Information

Publication History

Received 27 April 2004

Accepted after resubmission 05 July 2004

Publication Date:
06 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

The objective of this study was to characterize the role of fibrinogen in stimulating expression of inflammatory chemokines in endothelial cells through NF-κB activation. Human umbilical vein endothelial cells (HUVEC) were exposed to fibrinogen up to 3,000 µg/ml, and NF-κB activation was assessed using electrophoretic mobility shift assay (EMSA). Fibrinogen exposure resulted in a concentration dependent increase in NF-κB activation that reached a maximum at 1,000 µg/ml after 4 hours and was sustained up to 24 hours. The effect was inhibited by antibodies to αvβ3 and α5β1 and by the GRGDS peptide, indicating integrin involvement. Preincubation with Mn2+ lowered the fibrinogen concentration-dependence, consistent with integrin activation. Supershift assays demonstrated involvement of the p50, p65 and c-Rel components of NF-κB. Fibrinogen exposure also resulted in up-regulation of expression of monocyte chemoattractant protein-1 (MCP-1) and of interleukin-8 as shown by RNase protection assays and by real-time RT-PCR. Increased secretion of MCP-1 was confirmed by ELISA. Parthenolide, an IκB kinase inhibitor, prevented up-regulation of MCP-1 by fibrinogen, linking this response to NF-κB activation. From our findings, we conclude that fibrinogen regulates NF-κB activation and expression of inflammatory chemokines in endothelial cells and may be involved in mediating inflammatory processes.

Keywords

Fibrinogen - NF-κB - chemokines - endothelial cells
 

  • References

  • 1 Conforti G, Dominguez-Jimenez C, Zanetti A. et al. Human endothelial cells express integrin receptors on the luminal aspect of their membrane. Blood 1992; 80: 437-46.
    PubMedGoogle Scholar
  • 2 Bini A, Kudryk BJ. Fibrinogen in human atherosclerosis. Ann N Y Acad Sci 1995; 748: 461-71 discussion 71-3..
    PubMedGoogle Scholar
  • 3 Danesh J, Collins R, Appleby P. et al. Association of fibrinogen, C-reactive protein, albumin, or leukocyte count with coronary heart disease: meta-analyses of prospective studies. JAMA 1998; 279: 1477-82.
    CrossrefPubMedGoogle Scholar
  • 4 Ross R. Atherosclerosis—an inflammatory disease. N Engl J Med 1999; 340: 115-26.
    CrossrefPubMedGoogle Scholar
  • 5 Shin WS, Szuba A, Rockson SG. The role of chemokines in human cardiovascular pathology: enhanced biological insights. Atherosclerosis 2002; 160: 91-102.
    CrossrefPubMedGoogle Scholar
  • 6 Gerszten RE, Garcia-Zepeda EA, Lim YC. et al. MCP-1 and IL-8 trigger firm adhesion of monocytes to vascular endothelium under flow conditions. Nature 1999; 398: 718-23.
    CrossrefPubMedGoogle Scholar
  • 7 Rollins BJ, Yoshimura T, Leonard EJ. et al. Cytokine-activated human endothelial cells synthesize and secrete a monocyte chemoattractant, MCP-1/JE. Am J Pathol 1990; 136: 1229-33.
    PubMedGoogle Scholar
  • 8 Zeiher AM, Fisslthaler B, Schray-Utz B. et al. Nitric oxide modulates the expression of monocyte chemoattractant protein 1 in cultured human endothelial cells. Circ Res 1995; 76: 980-6.
    CrossrefPubMedGoogle Scholar
  • 9 Lind L. Circulating markers of inflammation and atherosclerosis. Atherosclerosis 2003; 169: 203-14.
    CrossrefPubMedGoogle Scholar
  • 10 Tak PP, Firestein GS. NF-κB: a key role in inflammatory diseases. J Clin Invest 2001; 107: 7-11.
    CrossrefPubMedGoogle Scholar
  • 11 De Martin R, Hoeth M, Hofer-Warbinek R. et al. The transcription factor NF-kappa B and the regulation of vascular cell function. Arterioscler Thromb Vasc Biol 2000; 20: E83-8.
    CrossrefPubMedGoogle Scholar
  • 12 Bunce LA, Sporn LA, Francis CW. Endothelial cell spreading on fibrin requires fibrinopeptide B cleavage and amino acid residues 15-42 of the beta chain. J Clin Invest 1992; 89: 842-50.
    CrossrefPubMedGoogle Scholar
  • 13 Gimbrone Jr MA, Cotran RS, Folkman J. Human vascular endothelial cells in culture. Growth and DNA synthesis. J Cell Biol 1974; 60: 673-84.
    CrossrefPubMedGoogle Scholar
  • 14 Sitrin RG, Pan PM, Srikanth S. et al. Fibrinogen activates NF-κ- B transcription factors in mononuclear phagocytes. J Immunol 1998; 161: 1462-70.
    PubMedGoogle Scholar
  • 15 Mukaida N, Shiroo M, Matsushima K. Genomic structure of the human monocytederived neutrophil chemotactic factor IL-8. J Immunol 1989; 143: 1366-71.
    PubMedGoogle Scholar
  • 16 Shyy YJ, Li YS, Kolattukudy PE. Structure of human monocyte chemotactic protein gene and its regulation by TPA. Biochem Biophys Res Commun 1990; 169: 346-51.
    CrossrefPubMedGoogle Scholar
  • 17 Brink N, Szamel M, Young AR. et al. Comparative quantification of IL-1β, IL-10, IL-10r, TNFalpha and IL-7 mRNA levels in UV-irradiated human skin in vivo. Inflamm Res 2000; 49: 290-6.
    CrossrefPubMedGoogle Scholar
  • 18 Hehner SP, Hofmann TG, Droge W. et al. The antiinflammatory sesquiterpene lactone parthenolide inhibits NF-κ B by targeting the I κ B kinase complex. J Immunol 1999; 163: 5617-23.
    PubMedGoogle Scholar
  • 19 Byzova TV, Goldman CK, Pampori N. et al. A mechanism for modulation of cellular responses to VEGF: activation of the integrins. Mol Cell 2000; 06: 851-60.
    PubMedGoogle Scholar
  • 20 Herrick S, Blanc-Brude O, Gray A. et al. Fibrinogen. Int J Biochem Cell Biol 1999; 31: 741-6.
    CrossrefPubMedGoogle Scholar
  • 21 Hoffmann A, Levchenko A, Scott ML. et al. The IκB-NF-κB signaling module: temporal control and selective gene activation. Science 2002; 298: 1241-5.
    CrossrefPubMedGoogle Scholar
  • 22 Gnant MF, Turner EM, Alexander Jr HR. Effects of hyperthermia and tumour necrosis factor on inflammatory cytokine secretion and procoagulant activity in endothelial cells. Cytokine 2000; 12: 339-47.
    CrossrefPubMedGoogle Scholar
  • 23 Bootle-Wilbraham CA, Tazzyman S, Marshall JM. et al. Fibrinogen E-fragment inhibits the migration and tubule formation of human dermal microvascular endothelial cells in vitro. Cancer Res 2000; 60: 4719-24.
    PubMedGoogle Scholar
  • 24 Karin M, Lin A. NF-κB at the crossroads of life and death. Nat Immunol 2002; 03: 221-7.
    PubMedGoogle Scholar
  • 25 Scatena M, Almeida M, Chaisson ML. et al. NF-κB mediates αvβ3 integrin-induced endothelial cell survival. J Cell Biol 1998; 141: 1083-93.
    CrossrefPubMedGoogle Scholar
  • 26 Kadish JL, Butterfield CE, Folkman J. The effect of fibrin on cultured vascular endothelial cells. Tissue Cell 1979; 11: 99-108.
    CrossrefPubMedGoogle Scholar
  • 27 Watanabe K, Tanaka K. Influence of fibrin, fibrinogen and fibrinogen degradation products on cultured endothelial cells. Atherosclerosis 1983; 48: 57-70.
    CrossrefPubMedGoogle Scholar
  • 28 Kaplan KL, Mather T, DeMarco L. et al. Effect of fibrin on endothelial cell production of prostacyclin and tissue plasminogen activator. Arteriosclerosis 1989; 09: 43-9.
    CrossrefPubMedGoogle Scholar
  • 29 Qi J, Kreutzer DL. Fibrin activation of vascular endothelial cells. Induction of IL-8 expression. J Immunol 1995; 155: 867-76.
    PubMedGoogle Scholar
  • 30 Dejana E, Languino LR, Polentarutti N. et al. Interaction between fibrinogen and cultured endothelial cells. Induction of migration and specific binding. J Clin Invest 1985; 75: 11-8.
    CrossrefPubMedGoogle Scholar
  • 31 Suehiro K, Gailit J, Plow EF. Fibrinogen is a ligand for integrin α5β1 on endothelial cells. J Biol Chem 1997; 272: 5360-6.
    CrossrefPubMedGoogle Scholar
  • 32 Harley SL, Powell JT. Fibrinogen up-regulates the expression of monocyte chemoattractant protein 1 in human saphenous vein endothelial cells. Biochem J 1999; 341: 739-44.
    CrossrefPubMedGoogle Scholar
  • 33 Takagi J, Springer TA. Integrin activation and structural rearrangement. Immunol Rev 2002; 186: 141-63.
    CrossrefPubMedGoogle Scholar
  • 34 Takagi J, Petre BM, Walz T. et al. Global conformational rearrangements in integrin extracellular domains in outside-in and insideout signaling. Cell 2002; 110: 599-11.
    CrossrefPubMedGoogle Scholar
  • 35 Ueda A, Ishigatsubo Y, Okubo T. et al. Transcriptional regulation of the human monocyte chemoattractant protein-1 gene. Cooperation of two NF-κB sites and NF-κB/Rel subunit specificity. J Biol Chem 1997; 272: 31092-9.
    CrossrefPubMedGoogle Scholar
  • 36 Roebuck KA. Regulation of interleukin-8 gene expression. J Interferon Cytokine Res 1999; 19: 429-38.
    CrossrefPubMedGoogle Scholar
  • 37 Hipp MS, Urbich C, Mayer P. et al. Proteasome inhibition leads to NF-κB-independent IL-8 transactivation in human endothelial cells through induction of AP-1. Eur J Immunol 2002; 32: 2208-17.
    CrossrefPubMedGoogle Scholar
  • 38 Collins T, Cybulsky MI. NF-κB: pivotal mediator or innocent bystander in atherogenesis?. J Clin Invest 2001; 107: 255-64.
    CrossrefPubMedGoogle Scholar
  • 39 Aiello RJ, Bourassa PA, Lindsey S. et al. Monocyte chemoattractant protein-1 accelerates atherosclerosis in apolipoprotein E-deficient mice. Arterioscler Thromb Vasc Biol 1999; 19: 1518-25.
    CrossrefPubMedGoogle Scholar