Reduction of Monocyte Chemoattractant Protein-1 and Interleukin-8 Levels by Ticlopidine in TNF- Stimulated Human Umbilical Vein Endothelial Cells
Ticlopidine has no effect on NF-kB activation. HUVECs were incubated with different concentrations of ticlopidine, and 100 g/mL of aspirin was added as the control. Cells were stimulated with/without TNF- (10 ng/mL) for 30 minutes, nuclear and cytosolic proteins were collected. Briefly, the nuclear protein extracts were collected, and western blots were performed using anti-NFkB p65 and anti-B23 Ab (a). Cytosolic protein extracts were collected, and western blots were performed using anti-Ik-B- and -tubulin Ab (b). Lane 1, without TNF-; lane 2, TNF- (10 ng/mL) alone; lane 3, TNF- + aspirin (100 g/mL); lane 4, TNF- + ticlopidine (1 g/mL); lane 5, TNF- + ticlopidine (10 g/mL); lane 6, TNF- + ticlopidine (30 g/mL). Results are representative of one of three independent experiments.