On the day of experiment, the cells were treated with check details complexes at indicated concentrations for 1 h. An equivalent concentration of DMSO was used in the control culture. In all experiments, incubation with 2 mM N-acetylcysteine
(NAC) for 1 h was used as a reference control. After treatment, cells were collected, washed with PBS and incubated with the 5 μM H2DCF-DA at 37 °C for 30 min in the dark. Immediately after staining cells were collected and analyzed by flow cytometry (FACSCalibur; Becton–Dickinson, Mountain View, CA, USA). All results were processed by using CellQuest software (Becton–Dickinson). Results and discussion Chemistry We have prepared a two series of Cu(II) complexes with a substituted pyrazoles (1a–c), as depicted in Fig. 1. Complexes 2a–c of the general formula (CuLCl2) were obtained in reaction of ligands Selleck Staurosporine with CuCl2·2H2O (in a 1:1 molar ratio) click here in ethyl acetate. The complexes 3a–c were synthesized in molar ratio 2:1 giving ionic complexes of general formula [CuL2](ClO4)2 (Fig. 2). The details of synthesis, results of elemental analysis and characterization of complexes using IR, NMR and MS spectroscopy were described in
our previous articles (Miernicka et al., 2008; Budzisz et al., 2009, 2010). All complexes were recrystallized from DMF, but only compounds 2a–c yielded crystals suitable for X-ray diffraction. The complexes exhibit trigonal bipyramidal configuration at Cu(II) centre. Fig. 1 Structure of the ligands Fig. 2 Proposed structures of the 2a–c and 3a–c complexes SOD/CAT/GPx-like
activity Complexes 2a–c and 3a–c were investigated on their antioxidant cAMP activity. The SOD (SOD-1), GPx and CAT activities and moreover total antioxidative status (TAS) have been determined. The results were expressed as enhancement (in %) of antioxidant enzymes activity and TAS value in blood samples treated with Cu(II) complexes in comparison to antioxidant activity in control samples and are presented in Fig. 3. Fig. 3 The enhancement (in %, mean value + SEM) of antioxidant activity of catalase, superoxide dismutase (SOD), glutathione peroxidase (GPx) and total antioxidant status (TAS) value in blood samples treated with Cu(II) complexes (20 μg/mL of blood) in comparison with antioxidant activity in control samples Our data underline that addition of compounds in blood lead to statistically significant increase in enzymes activities in comparison to control samples. The differences between two groups (samples with synthesized compounds and control group) were calculated using t test for dependent samples. T test results indicate that activity of CAT, SOD, GPx and TAS value in all samples with metal complexes was statistically significant (p < 0.01) greater than in control samples. SOD-1 is metalloprotein that catalyze ‘dismutation’ reaction which detoxify superoxide radicals (O 2 •− ) (Ercal et al.
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