The apoptotic/necrotic changes in isolated human peripheral blood mononuclear cells (MNCs) subjected to hydrogen peroxide (HO), cyanate (NaOCN) and their combination were examined. The mitochondrial potential (ΔΨm), the activities of caspases (-2, -3, -6, -8 and -9) and the level of carbonyls and amino groups in proteins were determined and DNA fragmentation. Apoptotic or necrotic cells were identified by fluorescence microscopy using double staining with Hoechst 33258/propidium iodide. Treatment of MNCs with NaOCN (1 mmol/L and 2 mmol/L), alone and in combination with HO (100 μmol/L), led to a significant decrease in the content of amine groups and a significant increase in the carbonyl level of MNCs in comparison with the control. Measurements taken at three time points (30, 60 and 150 min) showed a significant decrease in ΔΨm in MNCs incubated with HO, cyanate and their combination. The highest decrease in ΔΨm was observed after 150 min, when a combination of NaOCN and HO was applied. We observed significant increases in the activities of caspases-2 and -3 in cells exposed to HO and the combination of NaOCN and HO. An increase in caspase-2 but not in caspase-3 activity was noted in cells incubated with cyanate. A significant increase in caspase-9 activity in MNCs was observed in all arrangements of tested compounds in comparison with the control. In HO-treated cells, a higher level of necrotic cells was noted in comparison to apoptotic cells, whereas carbamylation led mainly to apoptotic cell death. The combination of cyanate and HO increased the population of necrotic cells.
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