[Reactive nitrogen and oxygen species metabolism in rat heart mitochondria upon administration of NO donor in vivo]

Abstract

Some aspects of reactive nitrogen and oxygen species (RNS and ROS) metabolism in rat heart mitochondria under administration of different doses of nitroglycerine (NG) in vivo are discussed. It is shown that NG administration results in a dose-dependent increase in Ca2+-uptake in mitochondria, due to the dose-dependent inhibition of mitochondrial permeability transition pore (MPTP) in vivo and the activation of Ca2+-dependent mitochondrial NOS. It was shown that NOS activity increases in accord with the increase of Ca2+-uptake in mitochondria. The dose-dependent activation of nitratreductase is observed. However, nitrite production decreases dose-dependently, according to the change of NO2-/NO3- ratio on behalf of NO3-, the end product of NO transformations. The relation between nitrosylation of mitochondrial proteins with the nitrosothiols formation and nitrate production also changes towards NO3-, which shows the activation of oxidation reactions in heart mitochondria after NG administration. Accordingly, dose-dependent increase in lipid peroxidation (LP) products is shown, the hallmark of the membrane damage in mitochondria. It is established that the cause of oxidative stress, besides the dose-dependent increase in ROS production (hydroperoxide, superoxide and hydroxyl-radical), lies in the increase of free iron content, derived from the oxidation of mitochondrial iron-containing proteins. The iron interaction with hydroperoxide following Fenton reaction as well as free-radical decomposition ofperoxynitrite, derived from NO3- are the possible cause of manifold increase in ROS as well as LP production, and RNS oxidation to NO3-. Thus, NO-dependent MPTP blockage, due to NO synthesis in mitochondria in vivo, results in the activation of both constituents of NO-cycle: NOS-dependent, due to Ca2+-dependent activation of mitochondrial NOS, and nitrate-reductase-dependent, due to the increase in NO3- formation. However, increase in ROS production, augmented by the iron release, leads to the oxidative stress and the shift of RNS metabolism towards NO3- formation, in spite of the activation of nitrate-reductase-dependent pathway of NO-cycle. It is shown that reversible MPTP opening in vitro diminishes ROS production, whereas MPTP blockage by cyclosporine A restores the ROS formation to control level. Thus, MPTP-dependent inhibition of ROS overproduction both in vitro and in vivo, shows the importance of MPTP in the regulation of ROS and RNS metabolism in mitochondria.