Type I collagen induces tissue factor expression and matrix metalloproteinase 9 production in human primary monocytes through a redox‐sensitive pathway

• Published in: Journal of thrombosis and haemostasis 2008-09, Vol.6 (9), p.1586-1594
• Main Authors: POITEVIN, S., GARNOTEL, R., ANTONICELLI, F., GILLERY, P., NGUYEN, P.
• Format: Article
• Language: English
• Subjects: Acetophenones - pharmacology; Acetylcysteine - pharmacology; Base Sequence; Blotting, Western; Cells, Cultured; collagen I; Collagen Type I - physiology; Cytokines - physiology; DNA Primers; Electrophoretic Mobility Shift Assay; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Humans; Matrix Metalloproteinase 9 - biosynthesis; metalloproteinases; monocytes; Monocytes - enzymology; Monocytes - metabolism; NF-kappa B - metabolism; Oxidation-Reduction; Pyrrolidines - pharmacology; Reactive Oxygen Species - metabolism; Reverse Transcriptase Polymerase Chain Reaction; ROS; Thiocarbamates - pharmacology; Thromboplastin - metabolism; tissue factor
• ISSN: 1538-7933; 1538-7836; 1538-7836
• DOI: 10.1111/j.1538-7836.2008.03051.x

Background: Tissue factor (TF), the main trigger of coagulation cascade, is a major component of the atherosclerotic plaque. Matrix metalloproteinases (MMPs) are recognized as key mediators of extracellular matrix remodeling during inflammation. It was recently emphasized that both TF and MMP‐9 were overexpressed in atherosclerotic plaques, suggesting a role of both molecules in plaque instability and thrombogenicity. Objective: The present study was designed to determine whether human monocytes could co‐express TF and MMP‐9 when the cells interact with type I collagen, a major component of the extracellular matrix and atherosclerotic plaque. Methods: Human monocytes were isolated by elutriation and incubated in collagen I‐coated plates. Tissue factor and MMP‐9 expression were examined using real‐time reverse transcription‐polymerase chain reaction, flow cytometry, western blot and zymography. The activation of nuclear factor‐κ B (NF‐κB) and the role of reactive oxygen species (ROS) in TF and MMP‐9 production was studied using gel shift experiments, antioxidants pyrrolidine dithiocarbamate (PDTC) and N‐acetyl‐cysteine (NAC), and apocynin (a specific inhibitor of the NADPH oxidase). Results: Type I collagen induced TF expression and increased MMP‐9 production. In addition, the pro‐inflammatory tumor necrosis factor‐α (TNF‐α), produced in response to collagen I, increased MMP‐9 production. PDTC and NAC inhibited NF‐κB activation during monocyte interaction with collagen I. Finally, both antioxidants and apocynin decreased the expression of TF, TNF‐α, and MMP‐9. Conclusions: These results indicate a new mechanism in the monocyte expression of TF and MMP‐9 in response to collagen I involving a ROS‐dependent pathway linked to the activation of the NADPH oxidase.