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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)

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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
 

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