Corn Gibberellic Ketene Toxic Effect Of Leydig Cells

Zearalenone is a mycotoxin produced by fusarium fungi that are found widely in foods and animal feeds, and it can cause reproductive and developmental toxicity, immunotoxicity, it also have some effects on tumorigenesis, so it has a great potential threat to animal performance and human health. In China ZEA contamination is graveness, and the effect is more and more widely. In recent years, more and more attention has been paid on the toxic effects of ZEA to male reproductive system. The studies reveal that ZEA poisoning can lead male animals to hypermastia and get feminization, preputial edema, hyposexuality, orchiatrophy and reduce the quantity and quality of semen. But as yet, the male reproductive toxicity caused by ZEA especially on the molecular level has not absolutely cleared. To explore the male reproductive toxicity cused by ZEA further, on the ground of establishing in vitro models of zearalenone and Leydig cells, the study investigated the Leydig cells toxicity caused by zearalenone on the cell and molecular level.The exposure concentractions of ZEA were determined by MTT method; The effect of ZEA to testosterone secretion in primary cultured SD rat Leydig cells which eoxposed in10U/ml HCG and ZEA was determined by ELISA kit; The DNA damage effect of ZEA to Leydig cells was detected by Single-Cell Gel Electrophoresis; The effect of ZEA to Leydig cells apoptotic rate was determined by flow cytometry and by determining the expression of apoptosis related protein Bax and Bcl-2in Leydig cells using the method of western blot. The results showed that:①the motility rat of Leydig cells reduced as the increase of ZEA dosage, and there is a significant difference between each groups (P<0.05), the cells motility rat is54.49%when exposed inlOμg/ml ZEA,which is close to lethal concentration50(LD50), so we determined the exposure concentrations1,5,10,20μg/ml.②ZEA has a depressant effect on testosterone secretion, the testosterone secretion had a significant downward trend (P<0.05) as the increase of the exposure concentration (exposed in0μg/ml、1μg/ml、10μg/ml、20μg/ml ZEA for12h) and the exposure time (exposed in5μg/ml ZEA for0h、1h、6h、12h、24h)③ZEA has a DNA damage effect on Leydig cells, the comet rate increased when exposure concentrate increased, and the tail length and DNA content in tail have a upgrade trend too, every group have a significant difference(P<0.05) to control group beside the group1μg/ml.④ ZEA can induce Leydig cells apoptosis, the apoptotic rate and the protein express of Bax/Bcl-2were firstly and then decreased as the increase of ZEA exposure concentration, That is the low concentration of ZEA mainly induce cells apoptosis and the high concentration mainly induce cells death. The apoptosis rate and Bax/Bcl-2were increased (P<0.05) as the increase of ZEA exposure time.The results showed that ZEN exposure can reduce the motility and the ability of testosterone secretion, induce DNA damage,and lead apoptosis of Leydig cells.