ASTA-treatment partially reduced (23%) the H2O2 production observed in FA group as compared with PMA-control group (Fig. 3C). There was

an increase Bleomycin molecular weight of 97% in NO production after treatment with 0.3 mM of FA as compared with control group without LPS. Treatment of cells with ASTA in the FA group did not prevent the increase caused by the presence of FA. ASTA per se, raises nitric oxide production by 99% as compared with control group without LPS ( Fig. 3D). N-acetylcysteine (NAC) and BSA partially reduced the NO production induced by the FA mixture. To determine whether the increased levels of ROS induced by the FA mixture can modulate antioxidant status of cells, we evaluated the antioxidant enzyme activities after 24 h of treatment (Table 1). The FA mixture decreased the activity of CAT by 42% and increased the total-SOD activity by 27% as compared to the control group. The FA group with ASTA restored total-SOD activity to those of the control group, whereas CAT activity decreased by 71% and GR activity increased by 80% as compared with the control group. Among all front line antioxidant enzymes tested, total-SOD was increased by 52% and GR HDAC inhibitor activity was decreased by 28% due to ASTA treatment. Oxidative damages in biomolecules were also

modulated by the FA mixture. TBARS levels were dramatically increased by treatment of cells with a FA mixture (210%) and ASTA-treatment partially restored TBARS levels (112%) as compared with the control cells. Free protein SH-group was decreased in 69% in cells treated with FA. After treatment with 2 μM of ASTA a partial restoring of 41% was observed in thiol content groups. Carbonyl groups were not modulated by the treatment of cells with FA and ASTA (data not shown). A significant reduction in the content of GSH and GSSG of 73% and 35%, respectively was observed in lymphocytes treated with 0.3 mM

of the FA mixture when compared to the control DNA ligase group. This reduction was not prevented by ASTA addition (Fig 4). It has been postulated that FA may influence cells of the immune system, including lymphocytes by modifying cell-membrane composition (Fan et al., 2004 and Li et al., 2006), altering intracellular signaling pathways (Gorjao et al., 2007, Lee et al., 2004, Madani et al., 2001 and Mizota et al., 2009), proliferation capacity, interleukins release (Nunes et al., 2008, Sacerdote et al., 2005 and Verlengia et al., 2004), ROS production (Cury-Boaventura and Curi, 2005 and Stentz and Kitabchi, 2006; Otton et al., 2007), gene expression (Verlengia et al., 2004) and calcium mobilization (Otton et al., 2007). Overall, the mixture of FA used in the present study caused a marked increase in the production of superoxide anion, hydrogen peroxide and nitric oxide, which was accompanied by an increase in total-SOD activity and in levels of TBARS as well as a reduction of catalase, levels of free thiol groups and GSH content.