Genotoxic response and alteration of intracellular redox balance in Hep-2 cell line by exposure to Iprodione

CHAUFAN GABRIELA; RIOS DE MOLINA, MARÍA DEL CARMEN; MUDRI, MARTA DOLORES; GALVANO, CAMILA; ANDRIOLI, NANCY

Roma

Congreso; SETAC Europe 28th annual meeting, Responsible and Innovative Reseach for Environmental Quality; 2018

SETAC Europa

The use of fungicides represents one of the most important factors in the control of pests and diseases, which affects the production systems of fruits and vegetables. It is known that most fungicide residues remain stable in food for long periods of time, increasing exposure risk for the general population. The aim of the present study was to evaluate the oxidative damage, the antioxidant response and the genotoxic effect in a cell line (HEp-2) against the exposure of sublethal concentrations of the fungicide Iprodione. For this proposal, we determine the content of protein carbonyls as marker of oxidative damage, the equivalent content of glutathione (GSH), the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) and from detoxifying enzyme GSH-S-transferase (GST), in 3 concentrations of Iprodione (1.5, 7 and 25 μg/ml). The cell division index, the replication index, the frequency of chromosomal aberrations and micronuclei were also determined in the presence of 7.5; 17.5 and 25 μg/ml of Iprodione. The cells were cultured in minimal essential medium supplemented with 10% fetal bovine serum (v/v), penicillin (100 U ml), streptomycin (100 mg/ml), amphotericin B (2.5 mg/ml) in a humid environment with 5% CO2 (v/v), at 37 ºC. For the cytotoxicity assays, the cells were seeded in 96-well plates, for enzymatic determinations and protein damage in Petri dishes (7.5x106 cells) and for genotoxicity parameters in 6-well plates. From the MTT assays, the LC50 was determined (29.88 (25.98 - 34.37) μg/ml Iprodione). The SOD activity decreased significantly 40% (p < 0.05) at 25 μg/ml of Iprodione, while no effect on the activity levels of CAT and GST was observed. The content of protein carbonyls increased 30% (p < 0.001) at the highest concentration of Iprodione tested. In addition, it was observed that Iprodione induces tripolar and micronucleus divisions at 17.5 and 25 μg/ml and bridges with all concentrations tested. Both the index of division and the index of replication indicate that the cells maintain their proliferation capacity, which allows to studying the biomarkers of genotoxicity in this system. These results confirm that Iprodione produces genotoxicity and an alteration in the redox equilibrium at the concentrations tested, which indicates the potential risk of exposure to this xenobiotic.