Regulation of proteinase-activated receptor 1 by inflammatory mediators in human vascular endothelial cells.
Thrombin plays a critical role in haemostasis, inflammation, and cell proliferation, mediated by proteinase-activated receptor 1 (PAR-1; thrombin receptor). The physiological and pathological regulation of PAR-1 by inflammatory mediators has not yet been fully elucidated. The aim of this study is to investigate the effects of inflammatory mediators on mRNA and protein expression of PAR-1 in early passage human vascular endothelial cells. Endothelial cells were activated by inflammatory mediators, such as tumour necrosis factor alpha (TNFalpha), interferon gamma (IFN gamma), and bacterial substance lipopolysaccharide (LPS), and the PAR-1 expression was verified by flow cytometry or RT-PCR. By stimulating endothelial cells with TNFalpha, IFN gamma, and LPS, the PAR-1 expression on the cell surface remained almost unchanged for 48 h. After stimulation with 20-300 U/ml TNFalpha, the total cellular PAR-1 expression (both on cell surface and in the cytoplasm) significantly decreased at 24h and thereafter recovered to the basal level at 48 h. The stimulation with 100 U/ml TNFalpha transiently down-regulated the PAR-1 mRNA expression to approximately 0.3-fold of the basal level at 30 min, but it rebounded 3-fold above the basal level at 6h, and again decreased to 0.5-fold of the basal level at 12h, and finally returned to the basal level at 24h. In contrast, IFN gamma or LPS did not affect the PAR-1 mRNA expression.
Department of Internal Medicine I, National Defense Medical College, Tokorozawa, Saitama, Japan.
Tadashi
T
Shinohara
Kimihiro
K
Suzuki
Kunio
K
Takada
Makoto
M
Okada
Fumitaka
F
Ohsuzu
D000970:Antineoplastic Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D018613:Microscopy, Confocal; D044463:Receptor, PAR-1; D018179:Receptors, Thrombin / Q000096:biosynthesis / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D000970:Antineoplastic Agents; D008070:Lipopolysaccharides; D044463:Receptor, PAR-1; D018179:Receptors, Thrombin; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma