Osteoprotegerin upregulates endothelial cell adhesion molecule response to tumor necrosis factor-alpha associated with induction of angiopoietin-2.

OBJECTIVE Osteoprotegerin (OPG) and osteopontin (OPN) have been identified within unstable atherosclerosis and circulating concentrates have been linked to cardiovascular events. We studied the influence of OPG and OPN on endothelial adhesion molecule expression and monocyte binding. METHODS Resting or tumor necrosis factor (TNF-alpha) activated human endothelial cells were incubated with OPG (0, 0.5, 5, and 10 ng/mL) or OPN (0, 2.5, 10 and 50 nmol/L). The expression of endothelial genes and proteins was investigated with the Oligo GEArray microarray series, multiplexed gene expression analysis, flow cytometry, ELISA and immunohistochemistry. Monocyte-binding studies were carried out using fluorescently labeled THP-1 cells and analysed by flow cytometry. RESULTS OPG but not OPN stimulated a dose-dependent increase in the expression of intercellular adhesion molecule-1, vascular cell adhesion molecule-1 and E-selectin by endothelial cells in the presence of TNF-alpha (p<or=0.05) which was reflected by enhanced binding of THP-1 monocytes. In the absence of TNF-alpha, OPG had no significant effect on adhesion molecule expression but upregulated angiopoietin-2. When the induction of angiopoietin-2 was inhibited using interfering RNA the ability of OPG to upregulate adhesion molecules in the presence of TNF-alpha was abolished. OPN did not effect adhesion molecule expression by resting or activated endothelial cells. CONCLUSION OPG upregulates angiopoietin-2 in human endothelial cells sensitizing them to the effects of TNF-alpha. These findings suggest a mechanism by which OPG may stimulate inflammation in atheroma and thereby promote the progression and complications of atherosclerosis.