The protective effect of follicular fluid against the emerging mycotoxins alternariol and beauvericin

Abstract

Porcine granulosa cells were cultured in the absence or presence of 10% porcine follicular fluid (FF) at different concentrations (0-20 mu M) of the mycotoxins alternariol (AOH) and beauvericin (BEA). The analyses were performed after exposure to these mycotoxins in a medium supplemented or not with FF harvested from gifts and sows. ,Cell enzymatic activity and nuclear membrane integrity were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and ethidium homodimer-1 labelling. Trolox equivalent antioxidant capacity was measured to calculate the capacity of the cells to counteract reactive oxygen species. qRT-PCR was used to determine the relative gene expression of efflux transporters (ABCG1 and ABCG2) as well CYP11 and CYP19. Mycotoxin cytotoxicity was more related to enzymatic activity than to nuclear membrane damage and no direct relationship with oxidative stress was observed, except when cells were exposed to AOH. In this case, medium supplementation with FF from sows increased the antioxidant capacity of the cells. AOH did not regulate gene expression in the present conditions, but 5 mu M BEA led to the up-regulation of ABCG2 gene expression and a down-regulation of CYP19 expression. In conclusion, follicular fluid from sows is capable to decrease toxicity of AOH and of BEA.