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

Min Guo; Sanjeev K. Sahni; Abha Sahni; Charles W. Francis

doi:10.1160/TH04-04-0261

Thrombosis and Haemostasis, Inhaltsverzeichnis

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

¹Hematology-Oncology Unit, Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York, USA

,

Sanjeev K. Sahni

¹Hematology-Oncology Unit, Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York, USA

,

Abha Sahni

¹Hematology-Oncology Unit, Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York, USA

,

Charles W. Francis

¹Hematology-Oncology Unit, Department of Medicine, University of Rochester School of Medicine and Dentistry, Rochester, New York, USA

› Institutsangaben

Abstract

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 α_vβ₃ 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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Referenzen

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