Severe mushroom toxin alpha amanitin causes generation of reactive oxygen species in liver tissues of mice - a comparative study by two different instrumental methods.

Alpha amanitin is a natural hepatotoxin isolated from deadly poisonous Amanita phalloides mushroom and causes dramatic toxic consequences mainly within the liver and kidneys. By our formerly ex vivo studies using a spectrophotometrical method we demonstrated that alpha amanitin can induce increased levels of lipid peroxidation (LPO) products in livers of mice treated by the toxin compairing to those of the control mice. We assumed that during its metabolism alpha amanitin probably is involved in free radical reactions which were the reason for developing of lipid peroxidation process in livers of mice. The aim of the present study was to evaluate by two different unstrumental methods: Electron Paramagnetic Resonance (EPR) direct and indirect spectroscopy and visible spectrophotometry, the effect of alpha amanitin on livers of mice treated by the toxin. Twenty hours after toxin treatment, ROS production marked by EPR spectra signals of spin-adducts in the mice livers could be detected by indirect EPR spectroscopy. It was found twice higher levels of ROS production in the livers of poisonous mice comparing to those of the control mice. By direct EPR spectroscopy higher levels of ascorbate free radicals were found in liver homogenates of treated mice in comparison with those of the control mice, which confirmed that an oxidative process was developed in livers of poisonous mice. At the same time the levels of lipid peroxidation products measured by visible spectrophotometry as malondialdehyde reactive products in livers of treated mice were slightly increased in comparison with those of the controls.