Antioxidant activity of micronized diosmin on oxygen species from stimulated human neutrophils

Flavonoids are known to reduce reactive oxygen species released by polymorphonuclear neutrophils (PMNs) in vitro. We have studied the effects of S5682 (Daflon 500 mg), a purified flavonoid fraction composed of 90% diosmin and 10% hesperidin. S5682 produced a dose-dependent inhibition of the luminol chemiluminescence (CL) induced by phorbol myristate acetate on PMNs (IC50 = 5 x 10(-5) M), with no effect on superoxide anion (O2.-) formation and on cellular superoxide dismutase activity as determined by lucigenin-amplified CL. The CL results were confirmed by the hydrogen peroxide (H2O2) determination showing that S5682 reduced H2O2 formed through either PMN stimulation (IC50 = 1.6 x 10(-6) M) or an in vitro enzymatic mechanism (IC50 = 2 x 10(-6) M). S5682 inhibited luminol-dependent CL induced by H2O2 (IC50 = 5 x 10(-6) M). However, O2 was not formed from H2O2 in contact with S5682 and the UV spectrum of this compound was not modified. In contrast, S5682 inhibited luminol-dependent CL induced by H2O2 in the presence of horseradish peroxidase (IC50 = 3 x 10(-6) M), and the UV spectrum of S5682 was modified. Luminol-dependent CL induced by hypochlorite (OCl- 10(-5) M) was also inhibited by S5682 (IC50 = 7 x 10(-5) M). This inhibitory effect was similar to that of sodium azide on myeloperoxidase activity. Moreover, OCl- 5 x 10(-4) M also altered the UV spectrum of S5682 10(-4) M. These results indicate that S5682 could be active on the H2O2-OCl(-)-myeloperoxidase system.