The effect of copper acetate on hepatocyte metabolism in vitro

Background . Copper ions are necessary for maintaining basic physiological processes in the mammalian organism. However, their excessive absorption or accumulation in cells can lead to the development or exacerbation of various pathological processes. The cytotoxic and genotoxic effects of high concentrations of copper compounds are currently well studied in various cell cultures, whereas the effect of non-toxic amounts of copper ions on physiological processes in cells, including during their cultivation, has been extremely poorly studied. The aim of the study . To investigate the effect of copper ions on changes in the intracellular amount of mitochondrial cytochrome C oxidase and glutathione synthetase. Materials and methods . A primary culture of hepatocytes was obtained, which was exposed to copper acetate at a concentration of 200 µg/ml in terms of copper content for 24 hours. After fixation, the samples were stained immunocytochemically using antibodies to cytochrome C oxidase (CcO) subunit I and glutathione synthetase (GS). Results . In hepatocyte culture, a significant increase in the intensity of fluorescent staining of the two analyzed enzymes was demonstrated both after 6 hours and after 24 hours of exposure to copper ions, which indicates a change in their number in cells. At the same time, the increase in the amount of CcO was more intense in the first 6 hours of incubation with a microelement, whereas in the next 18 hours, changes in the intracellular content of CcO were less pronounced. The increase in the intensity of the GS fluorescent stain was more active and was observed throughout the entire cultivation period. Conclusion . From the results obtained, it can be concluded that copper ions in non-toxic concentrations are able to influence key indicators of cell viability in culture by changing the amount of one of the main energy metabolism enzymes and the enzyme that provides synthesis of the most important low-molecular antioxidant glutathione.