Deoxynivalenol induces apoptosis in cells cultures and the antibiotic fosfomycin protects cells from nuclear changes.

PÉREZ, S.E.; MARTINEZ, G; RENSETTI, D; FERNANDEZ PAGGI, B; RICCIO, B; PEREZ, S. E; TAPIA, O; SORACI, A

Florida

Simposio; Global Veterinary Summit; 2015

Deoxynivalenol (DON) is a mycotoxin which inhibits protein synthesis and causes cytotoxicity. On the other hand, Fosfomycin (cis-1,2-epoxyphosphonic acid) (FOS) is a bactericidal broad-spectrum antibiotic used in human and veterinary medicine, which inhibits cell wall synthesis. It has been shown that FOS has some extra antimicrobial properties as phagocytosis promotion, immunomodulatory effects and protection against the toxicity caused by other drugs, among others. On previous studies we have demonstrated that after 4 h of incubation FOS (550 µg/mL) was able to prevent the cytopathic effect of DON (2.8 µg/mL) on Hep2 cells. To corroborate these results and to determine whether the effect of DON was due to the induction of apoptosis, Hep2 cells were seeded in 6 well-plates at a density of 4 x105 cells/well, and after 24 hours, cell monolayers were treated with DON at 2.8 µg/mL or with DON (2.8 µg/mL) and FOS (550 µg/mL). Four hours after treatment, cells were fixed with 4% paraformaldheyde and stained with 4´,6-diamino-2-phenylindole (DAPI). The presence of nuclear morphological changes representative of apoptosis (nuclear condensation and chromatin fragmentation) was evaluated under immunofluorescence microscope. Three to 6 visual field were observed, the percentage of apoptotic cells was determined and the results were evaluated by ANOVA/Tukey test. Experiments were assayed by triplicate. Cell cultures incubated with 2.8 µg/mL DON and 550 µg/mL FOS showed similarity to control culture plates (without antibiotic or toxin) showing the absence of cytotoxicity. Apoptotic cells percentage was 5.64 + 0.27 for the cells treated with DON and 0.65 + 0.5 for the cells incubated with both the mycotoxin and the antibiotic. The ANOVA/Tukey test HSD analysis shows a p-value of 0.0010053 (p