Figure 6
Lovastatin inhibits endotoxin-induced IL-8 release and superoxide production. Freshly isolated PBMC (~1.0 × 106 total cells/well, 100,000 monocytes) were prepared in 200 μl of incubation buffer with and without 1.5 μM lovastatin, and samples were incubated at 37°C for 16 h. (A) Cells were pelleted, washed gently, and resuspended in 200 μl of incubation buffer containing 100 ng/mL of LBP, and the indicated concentration of endotoxin. Cultures were then incubated at 37 °C for 6 h. At the end of the incubation period, cells were pelleted, and buffer samples were frozen at -20°C for subsequent IL-8 analysis by ELISA. Data are expressed as the mean ± S.E.M. of triplicate samples. (B) Cells were pelleted, washed gently, and resuspended on ice in individual wells of a 96-well white Optiplate in 200 μl of HBSS/0.1% HSA/0.1% G containing 100 ng/mL of LBP, 0.1 mM lucigenin, and the indicated concentration of endotoxin. The plates were then placed in the luminometer and two-sec readings were taken at 37°C every 2 min for 180 min. The resulting readings in relative light units (LU) were summed to provide a measure of activity. Data are expressed as total LU (in thousands) per 100,000 monocytes and represent the mean ± S.E.M. of 4 replicate samples; error bars for some conditions were so small they are obscured by the symbols. (A) **p < 0.001 for the no lovastatin control relative to the lovastatin value at the same LPS concentration. (B) *p < 0.05, **p < 0.001 for the no lovastatin control relative to the lovastatin value at the same LPS concentration. Similar results to A and B were seen in 3 independent experiments.