Study of Possible Mechanisms Involved in the Inhibitory Effects of Coumarin Derivatives on Neutrophil Activity
Phosphorylation of the NADPH oxidase component p40phox 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 level p40phox subunit was analyzed by Western blotting and detected by Phospho-p40phox (Thr 154) Antibody. The results are evaluated as optical density of p40phox corrected to optical density of the corresponding β-actin band. Mean ± SEM, (the data are representative of 4 donors performed in 2 separate experiments).