Human Aortic Endothelial Cells and Enhances Monocyte Adhesion Homocysteine Upregulates Vascular Cell Adhesion Molecule-1 Expression in Cultured

Elevated plasma homocysteine is an independent risk factor for atherosclerosis. We hypothesized that homocysteine enhances monocyte/human aortic endothelial cell (HAEC) interactions, a pivotal early event in atherogenesis, by upregulating endothelial adhesion molecules. After incubation of cultured HAECs with reduced DL-homocysteine for up to 24 hours, adhesion of human monocytes to homocysteine-stimulated HAECs was significantly upregulated in a timeand dose-dependent fashion. Pretreatment of HAECs with 100 mol/L homocysteine caused a 4.5-fold increase in the adhesion of normal human monocytes (P 0.001). Similarly, adhesion of monocytic U937 cells was maximally elevated by 3.5-fold at 100 mol/L homocysteine (P 0.001). In support of our hypothesis, vascular cell adhesion molecule (VCAM)-1 mRNA expression increased 5-fold in HAECs after 3 hours of treatment with 100 mol/L homocysteine, as assessed by quantitative reverse transcription–polymerase chain reaction. Neutralizing antibody studies confirmed the involvement of VCAM-1 in mediating monocyte adhesion to homocysteine-stimulated HAECs. Coincubation of HAECs with homocysteine and tumor necrosis factorsynergistically elevated monocyte adhesion as well as VCAM-1 protein expression, with the latter evaluated by flow cytometry. Preincubation of HAECs with cyclooxygenase inhibitors completely abrogated homocysteine-induced monocyte adhesion, whereas scavenging reactive oxygen species and the elevation of NO caused partial inhibition only. These data support the notion that the proinflammatory effects of homocysteine may have important implications in atherogenesis. (Arterioscler Thromb Vasc Biol. 2002;22:587-592.)