The flavonoids present in extract from Radix Scutellariae baicalensis Georgi are very effective antioxidants. It was proved that xylene has an influence on hydroxyl radical generation, increasing its level.

The aim of present study was to explain whether antioxidative properties of Antoxid (AX) are connected with its influence on hydroxyl radical generation and whether the mechanism connected with ^˙OH generation is responsible for biological effect of AX. This radical is very aggressive and could initiate lipid peroxidation. It isn't deactivated with superoxide dismutase or scavenge by other active molecule. So it was interesting to look into AX mechanism of inhibition of lipid peroxidation caused by xylene.

The study was performed on in vitro model, human placental mitochondria. The mitochondria were isolated by Radi method from mature placenta obtained after physiological delivery from Medical University Clinic. The proteins in mitochondria were measured by Lowry method. The Antoxid was dissolved in mitochondrial buffer (TRIS-HCl - pH-7.4) and used in following concentrations: 1.5, 3.0, 6.0, 12.0 and 30 µg/mL. The hydroxyl radical was measured by deoxyribose degradation.

At first the effect of AX on ^˙OH generation in mitochondria by t-BOOH was measured. The statistically significant (p<0.001) decrease in ^˙OH level was noted after AX treatment in doses 1.5-12.0 µg/ml. It looks, that inhibiting effect of AX on lipid peroxidation provoked by t-BOOH, expressed as MDA level, could be caused by inhibition of hydroxyl radical generation by AX. Quite different seems the mechanism of AX effect towards lipid peroxidation provoke by xylene. It looks that the pathway connected with ^˙OH generation is not responsible for MDA decrease resulted after AX treatment. When mitochondria exposed to xylene in dose 17.64 µg/mL were treated with AX in doses 1.5, 3.0, 6.0 or 12.0 µg/mL any decrease in ^˙OH generation was observed. Quite apposite the low doses (1.5 and 3.0 µg/mL) even stimulated the ^˙OH formation, whereas higher doses (6.0 or 12.0 µg/mL) don't show any influences.

The mechanism of AX preventing action was also examined by treatment of mitochondria with AX, 30 min. before exposition to xylene. The results demonstrate that also the preventing activity towards lipid peroxidation caused by xylene is not connected with inhibition ^˙OH generation by AX. The results show that ^˙OH generation was even increased by AX, but it didn't correlate with MDA increase.

Conclusions.

1. Antoxid decreased hydroxyl radical generation in exposure to t-BOOH but not to xylene.

2. The preventing and repairing effect of Antoxid in oxidative stress caused by xylene is not connected with hydroxyl radical scavenging.

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