As compared with the control selleckchem group, MPO activity was increased by 40% in the GM-group and reduced 86% and 94% in the AV and AVGM groups, respectively. Neutrophils were stimulated to produce hypochlorous acid by the addition of PMA (60 ng/well). Hypochlorous acid concentration was significantly reduced by 25% in the AV group and increased by 135%
and 99% in the GM and AVGM groups, respectively, when compared with the control group (Table 1). The maximum G6PDH activity was assessed by the reduction of the co-factor NADP+ into NADPH in human neutrophils (Table 1). GM promoted a significant reduction of 37% in G6PDH activity and astaxanthin + vitamin C addition (AVGM group) increased the G6PDH activity by 52% when compared to the GM group. TNF-α, IL-1β, and IL-6 are inflammatory cytokines which play important roles in immune responses to a variety of inflammatory stimuli. Therefore, we evaluated the effects of GM on TNF-α, IL-1β, and IL-6 after 18 h of LPS-stimulation. The levels of these cytokines LBH589 in the culture supernatants were measured using ELISA kits. Control neutrophils treated with LPS showed a significant increase in cytokine production when compared with the basal condition (100 ± 10 pg/ml, data not shown). The production of pro-inflammatory cytokines IL-6, IL-1β and TNF-α by human neutrophils in the AVGM group
was significantly decreased by 46%, 36% and 77%, respectively, when compared with the GM-group. IL-1β and TNF-α were also reduced in the AV-group by 42% and 89%, respectively, when compared with the control group. The production of reactive oxygen species is among the key weapons used by neutrophils to exterminate pathogens. In order to evaluate some possible modulation of
MGO + glucose and astaxanthin and vitamin C in a few of these species we used different probes. Superoxide anion production was measured by using two different probes, DHE and lucigenin. As assayed by the DHE probe, when GM-treated cells were stimulated with PMA there was an increase of 41% in the superoxide anion production compared with the PMA-control cells. Cells treated with astaxanthin plus vitamin C decreased production of superoxide anion by 54% as compared with the control-stimulated group. Addition of antioxidants to cells treated find more with GM (AVGM group) promoted a reduction of 66% in superoxide as compared with the GM group in stimulated conditions. Rotenone + Sodium Azide and DPI were added to neutrophils under PMA-stimulation. Both inhibitors significantly reduced superoxide anion production to basal levels. SOD enzyme addition was used to evaluate the specificity of DHE probe to superoxide anion (Fig. 3A), and as expected there was no significant fluorescence in this group. As an internal control, we also carried out the addition of 50 μM of H2O2 to PMA-treated cells. As expected, there was no increase in the fluorescence produced, thus ensuring the specificity of DHE for superoxide anion (data not shown). The lucigenin probe (Fig.