© 2003 by European Society of Cardiology
Copyright © 2003, European Society of Cardiology
Induction of matrix metalloproteinases-14 and -2 by cyclical mechanical stretch is mediated by tumor necrosis factor-
in cultured human umbilical vein endothelial cells
aDivision of Cardiology, Shin Kong Wu Ho-Su Memorial Hospital, 95 Wen-Chang Rd., Taipei 111, Taiwan
bDepartment of Emergency Medicine, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan
cGraduate Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan
* Corresponding author. Tel.: +886-2-2833-2211; fax: +886-2-2836-5775. shyukg{at}ms12.hinet.net
Objective: Mechanical forces have profound effects on endothelial cells. This study was undertaken to examine the hypothesis that tumor necrosis factor-
(TNF-) is a potential mediator of stretch-induced effects on matrix metalloproteinase (MMP). Methods: Human umbilical vein endothelial cells (HUVECs) grown on a flexible membrane base were stretched by vacuum to 20% of maximum elongation, at 60 cycles/min. We used the TNF- monoclonal antibody and c-Jun N-terminal kinase (JNK) inhibitor, SP600125, to investigate the cyclical stretch-induced expression of MMP-14 and -2 in cultured HUVECs. Results: Cyclical mechanical stretch significantly increased protein synthesis and mRNA expression for MMP-14 and -2 from 2 to 24 h. The increased MMP-14 and-2 proteins after stretch were completely blocked after the addition of TNF- monoclonal antibody (5 µg/ml) or SP600125 (20 µM) 30 min before stretch. By zymography, MMP-2 expression was induced by cyclical stretch and was attenuated by TNF- monoclonal antibody and SP600125. Cyclical stretch increased the immunohistochemical labeling of MMP-14 and -2 and significantly increased release of TNF- into the culture media from 120±2 to 331±2 pg/ml (P<0.001) after stretch for 12 h. Cyclical stretch increased and SP600125 decreased the phosphorylated JNK. Gel-shifting assay showed that DNA–protein binding activity of AP-1 increased after cyclical stretch and TNF- monoclonal antibody and SP600125 abolished the binding activity induced by cyclical stretch. Conclusion: These findings indicate that cyclical stretch augments TNF- production and MMP genes expression in HUVECs. TNF- mediates the stretch-induced MMP genes expression, at least in part, through the JNK pathway.
KEYWORDS Cell culture; Cytokines; Extracellular matrix; Gene expression; Stretch
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