Abstract
Tumor necrosis factor-α (TNF-α), an early inflammatory mediator typically regulated by nuclear factor kappa B (NF-κB), plays a critical role in the development of cardiovascular dysfunction in sepsis. While several myocardial cell types synthesize TNF-α, the importance of the myocardial endothelium in sepsis-related cardiac cytokine production is unclear. To determine the role of the human coronary artery endothelial cell (HCA-EC) in the cytokine response to endotoxin we measured in vitro TNF-α synthesis, TNF-α mRNA, and the associated NF-κB response to LPS. To determine the magnitude of the HCA-EC response we assessed the TNF-α and NF-κB response to LPS in a human monocytic cell line (THP-1) as well. We observed an increase in supernatant TNF-α from LPS-stimulated HCA-EC (12 h) that was ablated by the proteosome inhibitor, ALLN (N-acetyl-Leu-Leu-norleucinal). Similarly, ALLN-sensitive TNF-α was produced by monocytes following LPS, although at concentrations 100-fold higher than HCA-EC. TNF-α mRNA from HCA-EC was detected at 60 min in LPS-stimulated cells, but not in unstimulated cells or cells pretreated with ALLN. NF-κB p50/p65 subunits were detectable in endothelial nuclear protein 60 min following LPS. In contrast, NF-κB subunits from monocytes were detected at 15 min. Also, while ALLN only attenuated endothelial NF-κB translocation, monocyte NF-κB translocation was completely inhibited. These data suggest endotoxin-stimulated human coronary endothelial cells express TNF-α, which is regulated in part by NF-κB activation, in a manner and degree distinct from human monocytes.
Original language | English (US) |
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Pages (from-to) | 349-354 |
Number of pages | 6 |
Journal | Shock |
Volume | 16 |
Issue number | 5 |
DOIs | |
State | Published - Nov 2001 |
Keywords
- Electrophoretic mobility shift assay
- Endothelium
- Human coronary endothelial cells
- Human monocyte
- Lipopolysaccharide
- N-acetyl-Leu-Leu-norleucinal
- RT-PCR
- Sepsis
ASJC Scopus subject areas
- Emergency Medicine
- Critical Care and Intensive Care Medicine