Toxic Effects Of Zearalenone On The Early Embryo Implantation In Sows

Zearalenone is a Fusarium produced mycotoxin which mainly destroy the mammalian reproductive system,especially in pigs.The synchronized development of the blastocyst and uterine luminal and the cross-talk between them are essential to the implantation process and the estrogen level is one of the important factors in this process.Zearalenone can destabilize the balance of endocrine system as an oestrogen-like hormone and damage the intrauterine milieu as mycotoxin,affect the embryo implantation,and reduce the farrowing rate.Since it was rarely reported.We use the early pregnancy swine as experimental object to explore the effect of ZEA on embryo implantation and its molecular mechanism.The four groups with 5 swine post mating were exposed to 0,1,2,10mg/kg feed ZEA in 7-14 days.After sampling electron we had done HE stain,electron microscopy,primary cell were cultured for detecting apoptosis,proliferation,oxidative stress and mitochondrial membrane potential with flow cytometry detection.Further on autophagy using Western Blot,mRFP-GFP-LC3 double fluorescence detection system experiment technology of autophagy marker genes LC3 and P62 and apoptosis pathway proteins Bcl-2/Bax and Caspase-3 respectively on the tissues and cells were studied.The main results were as follows:1.HE staining results for endometrial showed the hemorrhage,inflammation and the hemal wall attenuates,there was a trend toward increased the endometrial epithelial cell layer thickness.2.The ultrastructure of endometrial was observed under electron microscopy,found that the autophagosomes increased significantly while the mitochondria were damaged hardly with ZEA concentration enrichment.The results show that ZEA leads to increased autophagy in endometrial tissue.3.Primary cell exposed ZEA cause a decline in cell proliferation rate and increased cell death;Apoptosis detection results show that the number of apoptosis increased with the higher concentration;Have also taken place in the mitochondrial membrane potential flow;Oxidative stress change is not obvious.4.Western Blot results found that high concentrations of ZEA lead to Bcl-2/ Bax expression decreased,LC-3II,Cleaved Caspase-3 higher expression in endometrial tissue,but there is P62 does not decrease with the increase of LC-3 II,confirmed that ZEA lead to cell apoptosis and activate autophagy,but autophagy flow is not smooth.5.Using MRFFP-GFP-LC-3 double fluorescence systems to observe that autophagy increases with the treatment of ZEA,but autophagy flow were suppressed,this result further confirmed the histones.6.The final conclusion: ZEA exposed in sow embryos implantation process,result in endometrium inflammation,and through autophagy damage and Caspase-3 pathways destroy the embryo implantation within uterus environment balance,that is affect embryos implant success.