Study of Possible Mechanisms Involved in the Inhibitory Effects of Coumarin Derivatives on Neutrophil Activity
PKC phosphorylation in PMA stimulated human neutrophils treated with 10 and 100 μmol/L HHC or HHDC. Human isolated neutrophils ( cells) were incubated at 37°C with HHC or HHDC (10 or 100 μmol/L) for 1 min, prior to addition of PMA (0.15 μmol/L). Cell lysates were prepared, and the protein levels of PKC isoenzymes (α and βII) were analyzed by Western blotting and detected by Phospho-PKCα/βII (Thr638/641) Antibody. The data are evaluated as optical density of PKC corrected to optical density of the corresponding β-actin band. Mean ± SEM, (the data are representative of 4 donors performed in 2 separate experiments).