Biologia, Bratislava, 55/Suppl. 8: 81-85, 2000.

ISSN 0006-3088 (Biologia). ISSN 1335-6399 (Biologia. Section Cellular and Molecular Biology).

 

Full Paper

Biochemical variables of oxidative cell and tissue damage induced by phenytoin.

 

Jana Navarova1*, Zuzana Seemannova2, Eduard Ujhazy1, Ruzena Sotnikova1, Michal Dubovicky1, Sona Muchova2 & Katarina Horakova2

1 Institute of Experimental Pharmacology, Slovak Academy of Sciences, Dubravska cesta 9, SK-84216 Bratislava, Slovakia; fax: ++ 421 7 54775928, e-mail: exfajana@savba.sk

2 Department of  Biochemistry and Microbiology, Faculty of Chemical Technology, Slovak University of Technology, Radlinskeho 9, SK-81237 Bratislava, Slovakia

* corresponding author

Received: July 24, 2000 / Accepted: October 24, 2000

 

Abstract

The aim of our study was to evaluate a model for studying possible preventive effects of selective natural and synthetic antioxidants on phenytoin (PHT)-induced biochemical changes in cells (in vitro) and in organs (in vivo). We compared the cytotoxic effect of PHT on HeLa cells using one direct (cell number) and four indirect (Lowry, MTT, NR, KB) methods. Further the adverse effect of PHT on pregnant Wistar/DV rats was studied using the activity of the lysosomal enzyme N-acetyl-ß-D-glucosaminidase (NAGA) and the level of glutathione (GSH)  in maternal serum, heart and aorta, foetal liver and brain, and in the placenta as markers of tissue damage. In vitro experiments: PHT caused unbalanced growth accompanied with increased production of proteins in the cells. The activity of lysosomal enzymes and mitochondrial dehydrogenases was increased. The amount of lysosomes in the cells was also raised. According to its cytotoxic activity, PHT can be classified as a toxic drug as it caused 53.5% inhibition of cell proliferation in 1mM concentration. In vivo experiments: On day 20 of gestation, PHT-induced toxic damage (150 mg/kg) was associated with an increase in NAGA activity in maternal serum and  in placenta  and foetal liver. The GSH level decreased in maternal aorta, in placenta and in foetal liver. NAGA activity and GSH level in maternal heart and in foetal brain remained unchanged. The PHT-induced biochemical changes established in the in vitro and in vivo models used provide the possibility to assess potential preventive effects of antioxidants.

 

Key words: phenytoin, oxidative stress, HeLa cells, NAGA, GSH, pregnant rats.