Toxicity mechanism of zearalenone on goat testicular interstitial cells

【Objective】To explore the effects of zearalenone(ZEA) on the viability,morphology and apoptosisrelated gene expression of goat testicular interstitial cells,and clarify the mechanism of its toxicity to goat testicular interstitial cells,providing theoretical basis for the subsequent discovery of the toxic mechanism of ZEA on the reproductive system of male goats.【Method】Goat testicular interstitial cells were treated with different concentrations of ZEA for 24 h. The cell proliferation was detected by CCK-8 method and the optimal concentration was selected. Then,the morphological characteristics,ultrastructural changes,damage healing ability,apoptosis and oxidative stress-related genes expression,mitochondrial membrane potential and intracellular reactive oxygen species(ROS) levels of testicular interstitial cells treated with ZEA were detected by optical microscopy,transmission electron microscopy,cell scratch test,realtime fluorescence quantitative PCR,JC-1 kit and ROS kit.【Result】With the increase of ZEA concentration,the viability cells died,the cells were wrinkled and bead-like,and the intercellular connections were loose,nuclear membrane was damaged,the endoplasmic reticulum was broken,the structure was blurred,the mitochondria was broken,autophagy vesicles were formed,and the nucleus was wrinkled and deeply stained. The cell healing ability was decreased,and the cell mobility was about 80% after 18 h of scratch culture. The relative expressions of Bcl-2,caspase-3 and SOD2 genes were significantly increased(P<0.05),and the relative expressions of Bax,HO-1,Nrf2,GPX1 and CAT genes were extremely significantly increased. JC-1 detection showed that the amount of red fluorescence decreased and the fluorescence intensity was weak,while the amount of green fluorescence increased and the fluorescence intensity was strengthened. ROS detection showed that the level of ROS in cells was significantly increased,that was,the degree of cellular oxidative stress was intensified.【Conclusion】ZEA exerts its toxic effect by inducing the deformation,mitochondria,endoplasmic reticulum and other cell ultrastructural damage of goat testicular interstitial cells,reducing their damage repair ability,the reduction of mitochondrial membrane potential and aggravating oxidative stress. of goat testicular interstitial cells decreased extremely significantly in a dose-dependent effect(P<0.01,the same below), and the half-inhibitory concentration of(IC₅₀) ZEA was around 20 μmol/L,so 20 μmol/L was selected as the treatment concentration for subsequent tests. After being treated with 20 μmol/L ZEA,a large number of goat testicular interstitial