Study of Possible Mechanisms Involved in the Inhibitory Effects of Coumarin Derivatives on Neutrophil Activity
Doses of the compounds tested producing 50% inhibition of control extracellular CL and intracellular CL of human neutrophils and cell-free CL system.
Human isolated neutrophils (5 × 105/sample) were stimulated with PMA (0.05 μmol/L) at 37°C, in the presence of HHC or HHDC (0.01–100 μmol/L). Extra- and intracellular ROS production were measured by isoluminol-/luminol-enhanced CL. In the cell-free system (luminol, horseradish peroxidase, and hydrogen peroxide), CL responses were measured in the presence of HHC or HHDC (1, 10, or 100 μmol/L) at 37°C. Percentage inhibition was calculated on the basis of integrated values of CL over 1800 s (extra- and intracellular CL) and of cell-free CL over 10 minutes. Mean ± SEM, –8. Control values given in RLU × seconds were 5.8 × 106 ± 1.4 × 106 for extracellular CL, 5 × 105 ± 0.5 × 104 for intracellular CL, and 3.8 × 105 ± 0.18 × 104 for cell-free CL.