南方农业学报2023，Vol.54Issue(11)：3396-3404,9.DOI:10.3969/j.issn.2095-1191.2023.11.026
玉米赤霉烯酮对山羊睾丸间质细胞的毒性机制研究
Toxicity mechanism of zearalenone on goat testicular interstitial cells
摘要
Abstract
[Objective]To explore the effects of zearalenone(ZEA)on the viability,morphology and apoptosis-related gene expression of goat testicular interstitial cells,and clarify the mechanism of its toxicity to goat testicular inter-stitial 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 morphologi-cal characteristics,ultrastructural changes,damage healing ability,apoptosis and oxidative stress-related genes expres-sion,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,real-time fluorescence quantitative PCR,JC-1 kit and ROS kit.[Result]With the increase of ZEA concentration,the viability 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 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 ex-tremely 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.关键词
山羊/玉米赤霉烯酮(ZEA)/睾丸间质细胞/细胞凋亡/线粒体膜电位Key words
goat/zearalenone(ZEA)/testicular interstitial cells/apoptosis/mitochondrial membrane potential分类
畜牧业引用本文复制引用
宁春妹,赵颖,安佳秀,付叶,田亚原,杨艺..玉米赤霉烯酮对山羊睾丸间质细胞的毒性机制研究[J].南方农业学报,2023,54(11):3396-3404,9.基金项目
国家自然科学基金项目(32002205) （32002205）
贵州省科技支撑计划项目(黔科合支撑[2021]一般154号) National Natural Science Foundation of China(32002205) （黔科合支撑[2021]一般154号）
Guizhou Science and Technology Support Plan Project(QKHZC[2021]154) （QKHZC[2021]154）
