摘要
为探究白藜芦醇(RSV)对玉米赤霉烯酮(ZEA)致睾丸支持细胞氧化损伤及凋亡的保护作用,以小鼠睾丸支持细胞系TM4细胞为研究对象,分别设对照组、ZEA组(20μmol/L)、RSV组(5μmol/L)、ZEA+RSV组(5μmol/L RSV处理30 min后+20μmol/L ZEA),24 h后,通过CCK-8法检测细胞活力,流式细胞术检测ROS水平和细胞凋亡率,试剂盒检测细胞内MDA含量和SOD活力,免疫荧光法结合流式细胞术检测线粒体膜电位(ΔΨm)变化,Western-blot检测Fas/FasL通路相关蛋白表达。结果显示,与对照组相比,ZEA极显著抑制TM4细胞活性(P<0.01),增加细胞凋亡率及提高ROS、SOD和MDA水平(P<0.01),并能够显著增加Fas/FasL信号通路相关蛋白FasL、FADD、Cleaved-caspase8的表达水平(P<0.05),极显著增加Fas的表达水平(P<0.01);与ZEA组相比,ZEA+RSV组细胞活力显著提高(P<0.05),细胞凋亡率和SOD水平显著降低(P<0.05),ROS和MDA水平极显著下降(P<0.01),Fas、FasL表达下调(P<0.01和P<0.05)。结果表明,RSV可有效减轻ZEA对小鼠睾丸支持细胞的氧化损伤作用,并能够通过调控Fas/FasL通路缓解ZEA诱导的睾丸支持细胞凋亡。
In order to explore the protective effect of resveratrol(RSV)on oxidative damage and apoptosis of testicular support induced by zearalenone(ZEA),TM4 cells(mouse testicular sertoli cell line)were divided into control group,ZEA(20μmol/L)group,RSV(5μmol/L)group and ZEA+RSV(5μmol/L RSV+20μmol/L ZEA)group.24 hours later,the cell viability was detected by CCK-8.The level of ROS and the rate of apoptosis were detected by flow cytometry.The intracellular MDA content and SOD activity were measured by using the kits,the changes of mitochondrial membrane potential(ΔΨm)were detected by immunofluorescence combined with flow cytometry,and the expression of proteins related to Fas/FasL pathway was detected by western blot.The results showed that compared with the control group,ZEA significantly inhibited the activity of TM4 cells(P<0.01),increased the apoptosis rate and the levels of ROS,SOD and MDA(P<0.01),and significantly increased the expression of Fas/fasL signal pathway related proteins Fas,FasL,FADD,Cleaved-caspase8(P<0.05 or P<0.01).Compared with ZEA group,the cell viability of ZEA+RSV group was significantly increased(P<0.05),while the apoptosis rate and SOD level were significantly decreased(P<0.05),ROS and MDA levels were significantly decreased(P<0.01),and Fas,FasL expression was down-regulated(P<0.01 or P<0.05).and Fas,FasL expression was down-regulated(P<0.01 or P<0.05).It is concluded that RSV can effectively reduce the oxidative damage of mouse testicular sertoli cells induced by ZEA and alleviate the apoptosis of testicular sertoli cells induced by ZEA by regulating Fas/FasL pathway.
作者
佘进进
冯楠楠
郑豪
刘双双
邹辉
顾建红
袁燕
刘学忠
刘宗平
卞建春
SHE Jin-jin;FENG Nan-nan;ZHENG Hao;LIU Shuang-shuang;ZOU Hui;GU Jian-hong;YUAN Yan;LIU Xue-zhong;LIU Zong-ping;BIAN Jian-chun(College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2020年第11期1453-1460,共8页
Chinese Veterinary Science
基金
国家重点研发计划支持项目(2016YFD0501208)
江苏高校优势学科建设工程资助项目(PAPD)。
作者简介
佘进进(1996-),男,江苏如皋人,硕士生,主要从事动物营养代谢病与中毒病研究,E-mail:mx120190774@yzu.edu.cn;通讯作者:卞建春(1965-),男,教授,博士,博士生导师,主要从事动物营养代谢病与中毒病研究,Tel:0514-87979042,E-mail:jcbian@yzu.edu.cn。
