Oxidative stress, uptake and bioconversion of 5-fluorouracil in algae

Impact of cytostatic drug 5-fluorouracil (FU) and its metabolite 2-fluoro-3-alanine (FA) on green alga Scenedesmus quadricauda was studied. FA elevated fluorescence signal of reactive oxygen species (ROS) more pronouncedly than FU at 1 and 10 μM doses while both ROS and reactive nitrogen species (RNS/NO) increased more expressively in 100 μM FU treatment. Cellular damage staining (Acridine Orange and Calcofluor White) did no reveal substantial difference between FU and FA. Majority of free amino acids including proline was unaffected after 24 h of exposure. FA depleted ascorbate peroxidase activity more than FU therefore ascorbate content (AsA) was less affected while FU stimulated glutathione reductase activity less than FA and therefore glutathione (GSH) was more depleted. Both compounds accumulated concentration-dependently with higher absolute FA amounts but FU conversion to FA was also detected. We subsequently influenced 100 μM FU- and FA-induced changes using known ROS (DTT - dithiothreitol) and RNS/NO (SNP - sodium nitroprusside and PTIO - 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide) modulators and results showed that PTIO depleted NO and elevated ROS while the opposite was found after SNP and DTT addition. Changes of lipid peroxidation (using BODIPY staining) confirmed that FU and FA toxicity is related to alteration of ROS/RNS balance.