Reduction of Monocyte Chemoattractant Protein-1 and Interleukin-8 Levels by Ticlopidine in TNF- Stimulated Human Umbilical Vein Endothelial Cells
Reduction of MCP-1 mRNA and protein levels by ticlopidine in HUVECs. HUVECs were incubated with serum-free medium for 12 hours before different concentrations of ticlopidine were added, after 12 hours incubation cells were stimulated with/without TNF- (10 ng/mL) for another 24 hours. (a) Cell growth was analyzed by MTT. Results are mean ± SEM (). (b) Total RNA was extracted and analyzed by RT-PCR. Lane 1, without TNF-; lane 2, TNF- (10 ng/mL) alone; lane 3, TNF- + ticlopidine (0.1 g/mL); lane 4 TNF- + ticlopidine (30 g/mL); M denotes molecular size marker. The lower panel is -actin as internal control. Results are representative of one of three independent experiments. (c) Relative amount of MCP-1 mRNA level was determined by quantitative real-time RT-PCR. (d) Culture supernatants were analyzed by ELISA. Data are expressed as the mean ± SEM of duplicate wells and are representative of five individual experiments. Significantly different versus TNF-treated alone *P < .05, **P < .001.