Effect of peroxynitrite on production of superoxide anion radicals and calcium homeostasis in cells of astroglial origin

Abstract

We studied the effect of a donor of peroxynitrite, SIN-1, on the morphological characteristics of interweaved rat С6 glioma cells, on menadione-induced production of superoxide anion radicals, and on the concentration of Са 2+ in these cells. In concentrations of 1.25·10 -4 to 2.5·10 -7 М, SIN-1 demonstrated cytotoxic and antimitogenic effects. This donor of peroxynitrite caused abnormal modifications of the size of С6 cells and the structure of cellular organelles, intensified in a dose-dependent manner the release of Са 2+ from cellular stores into the cytoplasm, and suppressed menadione-induced production of superoxide anion radicals. Therefore, it should be believed that peroxynitrite exerts a modifying effect on the processes of mitotic division and induces apoptosis; it is also involved in the processes of intracellular signalling providing an increase in the concentration of cytosolic Са 2+ and a decrease in the redox activity of cells.

Proliferation and Survival of Rat C6 Glioma Culture in the Presence of Implants Coated with Modified Carbon-Based Films

“…It is one of the most widely used strains, easily cultured in vitro, with large (up to 20-30 μ) structural elements [1]. Tumor gliocytes (10 5 cell/ml) were cultured in Petri dishes (35 mm) in MEM with 10% fetal calf serum and 10 -4 g/ml gentamicin sulfate.…”

“…Cell viability and number of dead tumor cells [1] were evaluated by staining with 0.2% trypan blue (Merck) in phosphate buffer saline (pH 7.2). The culture was visualized by computerized device for light microscopy.…”

“…The culture was visualized by computerized device for light microscopy. Mitotic activity (proliferation index) was calculated in a Goryaev chamber as the proportion of grown to inoculated cells [1]. Each studied group was represented by three independent inoculations.…”