Cylindrospermopsin-induced protein synthesis inhibition and its dissociation from acute toxicity in mouse hepatocytes.
The toxicology of the cyanobacterial alkaloid cylindrospermopsin (CYN), a potent inhibitor of protein synthesis, appears complex and is not well understood. In exposed mice the liver is the main target for the toxic effects of CYN. In this study primary mouse hepatocyte cultures were used to investigate the mechanisms involved in CYN toxicity. The results show that 1-5 microM CYN caused significant concentration-dependent cytotoxicity (52%-82% cell death) at 18 h. Protein synthesis inhibition was a sensitive, early indicator of cellular responses to CYN. Following removal of the toxin, the inhibition of protein synthesis could not be reversed, showing behavior similar to that of the irreversible inhibitor emetine. In contrast to the LDH leakage, protein synthesis was maximally inhibited by 0.5 microM CYN. No protein synthesis occurred over 4-18 h at or above this concentration. Inhibition of cytochrome P450 (CYP450) activity with 50 microM proadifen or 50 microM ketoconazole diminished the toxicity of CYN but not the effects on protein synthesis. These findings imply a dissociation of the two events and implicate the involvement of CYP450-derived metabolites in the toxicity process, but not in the impairment of protein synthesis. Thus, the total abolition of protein synthesis may exaggerate the metabolite effects but cannot be considered a primary cause of cell death in hepatocytes over an acute time frame. In cell types deficient in CYP450 enzymes, protein synthesis inhibition may play a more crucial role in the development of cytotoxicity.
Froscio
Suzanne M
SM
Department of Clinical and Experimental Pharmacology, University of Adelaide, Adelaide, South Australia 5005, Australia. suzanne.froscio@rmit.edu.au
Humpage
Andrew R
AR
Burcham
Philip C
PC
Falconer
Ian R
IR
D000470:Alkaloids; D000704:Analysis of Variance; D000818:Animals; D001427:Bacterial Toxins; D000087522:Cyanobacteria Toxins; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000737:chemistry; D051379:Mice; D010452:Peptide Biosynthesis / Q000187:drug effects*; D011335:Proadifen / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000633:toxicity*; D014498:Uracil / Q000031:analogs & derivatives* / Q000633:toxicity*
D000470:Alkaloids; D001427:Bacterial Toxins; D000087522:Cyanobacteria Toxins; D065607:Cytochrome P-450 Enzyme Inhibitors; D011500:Protein Synthesis Inhibitors; C089595:cylindrospermopsin; D014498:Uracil; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D011335:Proadifen; D007770:L-Lactate Dehydrogenase