Inhibitory effects of bucillamine on the expression of vascular cell adhesion molecule-1 in human umbilical vein endothelial cells.

Bucillamine (BUC) has been found to have beneficial effects in the treatment of rheumatoid arthritis (RA), in which the activation of endothelial cells plays an important role in the pathogenesis. The current studies examined the effect of BUC and its intramolecular disulfide form (BUC-ID) on the expression of adhesion molecules in human umbilical vein endothelial cells (HUVEC) stimulated with tumor necrosis factor-alpha (TNF-alpha). HUVEC (4 x 10(4)/well) were incubated with medium M199 containing heparin and 20% FCS with endothelial cell growth supplement (ECGS) for 24 h in the presence or absence of BUC or BUC-ID, after which the culture medium was replaced with ECGS free medium. Then the cultures were further carried out for additional 24 h with TNF-alpha (10 ng/ml) in the presence or absence of BUC or BUC-ID. BUC-ID, but not BUC, appeared to suppress the expression of VCAM-1 on HUVEC stimulated with TNF-alpha in a dose-response manner at its pharmacologically relevant concentrations (0.3-3.0 microg/ml), whereas only the 3 microg/ml concentration level of BUC-ID had a statistically significant effect, although the effect was relatively small. By contrast, lower concentrations of BUC-ID (1-3 microg/ml) suppressed the secretion of soluble VCAM-1 by HUVEC much more effectively. Of note, at the concentration of 3 microg/ml neither BUC nor BUC-ID significantly influenced the expression of ICAM-1 and E-selectin on TNF-alpha stimulated HUVEC. These results indicate that BUC-ID, but not BUC, specifically downregulates the surface expression of VCAM-1 as well as the release of soluble VCAM-1 by HUVEC stimulated with TNF-alpha. BUC-ID suppressed the production of solubleVCAM-1 by RA bone marrow CD34+ cells stimulated with SCF, GM-CSF and TNF-alpha more effectively than BUC. The data thus suggest that one of the mechanisms of action of BUC involves the inhibition of the activation of endothelial cells.