Inhibition of NiCl2-induced MCP-1 mRNA and protein expression by pharmacological blockade of p38 and PC-PLC.
(A-B) HUVECs were cultured for 8 hours in the absence or presence of 1.25, 5, or 20 μM SB202190 (A) or 1.56, 6.25, or 25 μg/mL D609 (B) and stimulated with 1.5 mM NiCl2 as indicated. Thereafter, HUVECs were processed for detection of MCP-1 mRNA by Northern blot analysis. To control equal RNA loading of Northern blot lanes, ethidium bromide stains of 18S and 28S rRNA are shown. (C-D) Supernatants from HUVECs exposed to 1.25, 5, or 20 μM SB202190 (C) or 0.39, 1.56, 6.25, or 25 μg/mL D609 (D) 30 minutes before and during a 12-hour period of stimulation with 1.5 mM NiCl2 were studied for MCP-1 content by ELISA. Data are expressed as mean ± SE of duplicate wells from 2 independent experiments.
