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METHODS
In vitro chemical assays
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Scavenging activity of antioxidant compounds (including plant extracts) against Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (RNS):
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Superoxide anion radical;
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Hydrogen peroxide;
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Hypochlorous acid;
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Singlet oxygen;
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Peroxyl radicals;
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Nitric oxide;
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Peroxynitrite.
Microplates used for the in vitro chemical assays
In vitro cellular assays
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Isolated human neutrophils:
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Viability studies: trypan blue and LDH assays;
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Evaluation of apoptosis by morphology and flow cytometry (annexin-V/PI);
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p53 determination;
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Measurement of caspases activity;
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Evaluation of the neutrophils' oxidative burst modulation;
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Neutrophils' chemotaxis assay;
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Inhibitory effect of the inflammatory enzyme 5-LOX;
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Determination of the enzymatic type of inhibition of soybean LOX-1.
Isolated human neutrophils
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Isolated human erythrocytes:
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Lipid peroxidation (TBARS);
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Hemoglobin oxidation;
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Hemolysis;
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Both forms of glutathione: reduced (GSH) and oxidized (GSSG).
Whole blood assays:
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Cytokines determination;
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Inhibitory effect of the inflammatory enzymes (COX-1 and COX-2).
Isolated human erythrocytes
(red portion)