Zinc protection of HepG2 cells from sporidesmin toxicity does not require de novo gene transcription.
Sporidesmin is an epidithiodioxopiperazine mycotoxin secreted by the saprophytic fungus Pithomyces chartarum. Ingestion of sporidesmin by ruminants grazing on the saprophyte infested pasture causes severe liver and bile duct damage leading to secondary photosensitisation. Zinc supplementation is used as an effective prophylaxis against sporidesmin toxicity in ruminants, however, the mechanism by which zinc protects is unknown. This study used the human hepatoma cell line, HepG2, as a model to examine the mechanism of zinc protection against sporidesmin toxicity. Treatment of cells with various concentrations of sporidesmin (0-10 microg/ml) resulted in a sigmoidal dose response curve with an LC50 of 5 microg/ml. Cells were protected from sporidesmin toxicity by pre-treatment for 2h or 16 h with zinc sulphate in a concentration dependent manner, with significant protection at 50 microM zinc and maximal protection at 200 microM zinc. To determine whether zinc protection required de novo gene transcription, cells were treated with the transcriptional inhibitor actinomycin D for one hour prior to and throughout the zinc pre-treatment. The presence of actinomycin D did not significantly reduce the zinc protection against sporidesmin cytotoxicity (80% protection without actinomycin D versus 71% protection with actinomycin D). Therefore, de novo gene transcription does not play a major role in the mechanism of zinc protection against sporidesmin toxicity in HepG2 cells.
Duncan
Elizabeth J
EJ
AgResearch Molecular Biology Unit, Department of Biochemistry, University of Otago, P.O. Box 56, Dunedin, New Zealand.
Thompson
Mary P
MP
Phua
Sin H
SH
D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects*; D003609:Dactinomycin / Q000494:pharmacology; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D020011:Protective Agents / Q000494:pharmacology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D013173:Sporidesmins / Q000633:toxicity*; D014158:Transcription, Genetic; D015032:Zinc / Q000494:pharmacology*
D020011:Protective Agents; D011500:Protein Synthesis Inhibitors; D013173:Sporidesmins; D003609:Dactinomycin; D007770:L-Lactate Dehydrogenase; C001873:sporidesmin; D015032:Zinc