Abstract
Zearalenone (ZEN), a widely known mycotoxin, is mainly produced by various Fusarium species, and it is a potent estrogenic metabolite that affects reproductive health in livestock and humans. In this study, the molecular mechanisms of toxicity and cell damage induced by ZEN in GC-1 spermatogonia (spg) cells were evaluated. Our results showed that cell viability decreased and apoptosis increased in a dose-dependent manner when GC-1 spg cells were exposed to ZEN. In addition, the key proteins involved in apoptosis, cleaved caspase-3 and -8, BAD, BAX, and phosphorylation of p53 and ERK1/2, were significantly increased in ZEN-exposed GC-1 spg cells for 24 h, and cytochrome c was released from mitochondria by ZEN. Interestingly, ZEN also triggered autophagy in GC-1 spg cells. The expression levels of the autophagy-related genes Atg5, Atg3, Beclin 1, LC3, Ulk1, Bnip 3, and p62 were significantly higher in ZEN-treated GC-1 spg cells, and the protein levels of both LC3A/B and Atg12 were remarkably increased in a dose-dependent manner in ZEN-exposed GC-1 spg cells compared to the control. In addition, immunostaining results showed that ZEN-treated groups showed a remarkable increase in LC 3A/B positive puncta as compared to the control in a dose-dependent manner based on confocal microscopy analysis in GC-1 spg cells. Our findings suggest that ZEN has toxic effects on tGC-1 spg cells and induces both apoptosis and autophagy.
Highlights
We evaluated the effects of ZEN in vitro using spermatogonia GC-1 spg cells, which were derived from 10-day mouse testes
The cell viability of GC-1 spg cells decreased in a dose-dependent manner when the cells were exposed to 30–300 μM ZEN for 24 h compared to the control (0 μM ZEN)
GC-1 spg cells is unclear, our results have clearly demonstrated the toxic effect of ZEN on spermatogonia via the study of apoptosis and autophagy mechanisms
Summary
Zearalenone (ZEN), known as the F-2 toxin, is a non-steroid mycotoxin with estrogenic effects produced by some Fusarium and Gibberella species [1]. ZEN can accumulate in the body and induce reproductive toxicity. In vivo studies have reported on the toxicological effects of ZEN in females, including functional alterations in reproductive organs, a depressing of the maturation of ovarian follicles, oocyte death in the follicles, a lack of ovulation [5,6], decreased fertility, increased embryo-lethal resorption, and abnormal hormone levels in the female reproductive system of rodents and livestock [7,8]. Male mice exposed to this mycoestrogen showed an increased number of abnormal spermatozoa; a low pregnancy rate was observed when females were mated with ZEN-exposed males [10]
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