摘要
为探讨自噬在玉米赤霉烯酮（ZEA）诱导睾丸支持细胞株TM4凋亡中的作用,用不同浓度ZEA（0、5、10、20μmol/L）处理细胞24 h后,流式细胞术检测细胞凋亡率;Western blot检测Bax等凋亡相关蛋白的表达情况;同时检测20μmol/L ZEA联合自噬抑制剂氯喹（CQ）或促进剂雷帕霉素（RAP）后对TM4细胞凋亡率和凋亡相关蛋白及自噬关键蛋白LC-3的变化。结果显示：与对照组相比,5μmol/L ZEA以上染毒组Bax/Bcl-2的比值、cleaved-caspase-3和cleaved-caspase-9蛋白表达量均显著或极显著增加。20μmol/LZEA＋c Q可使ZEA诱导的TM4细胞的凋亡率上升,Bax/Bcl-2的比率以及cleaved-caspase-3、cleaved-caspase-9蛋白表达显著下降而自噬标志蛋白LC-3的表达上升;20μmol/LZEA＋RAP,可使ZEA诱导的TM4细胞的凋亡率下降;Bax/Bcl-2的比率,cleaved-caspase-3和cleaved-caspase-9蛋白表达及LC-3的表达显著上升。结果表明：在ZEA诱导的TM4细胞发生凋亡的过程中,细胞凋亡率与自噬水平的高低有明显的关系,抑制自噬,ZEA诱导的TM4细胞凋亡率明显升高;促进自噬,ZEA诱导的TM4细胞凋亡率明显降低。研究表明,自噬能够延迟TM4细胞发生凋亡,自噬在ZEA对TM4细胞的毒性损伤中发挥保护作用。
In order to investigate the role of autophagy in the apoptosis of TM4 cells induced by zearalenone （ZEA）, TM4 cells were used as experiment model. After exposing TM4 cells to different concentrations of ZEA （0, 5, 10 and 20 μmol/L） for 24 h, the flow cytometry was used to detect the apopotic rate, and western blot was used to detect expression of apoptosis-related proteins. The changes in the apopto-sis rate, apoptosis-related protein and autophagy-related protein LC-3 in TM4 cells were detected by 20 μmol/L ZEA combined with auto-phagy chloroquine （CQ） or Rapamyein （Rap）. The results showed that, compared with the control group, the ratio of Bax/Bcl-2 and the expression of cleaved-caspase-3 and cleaved-caspase-9 were decreased and the expression of autophagy marker protein LC-3 was increased significantly （P〈0. 05 or P〈0. 01 ）. The apoptosis rate increased significantly after the ZEA and CQ, compared with that after the ZEA treat-ment. The ratio of Bax/Bcl-2, cleaved-caspase-3 and eleaved-easpase-9 and LC-3 protein expressions decreased significantly （P〈0. 05 or P〈0. 01 ）. The apoptosis rate significantly decreased after the ZEA and Rap, compared with that after the ZEA treatment. The ratio of Bax/ Bcl-2, cleaved-caspase-3 and cleaved-easpase-9 and LC-3 protein expressions increased significantly （P〈0. 05 or P〈0. 01）. The apop-totic rate was significantly correlated with the level of autophagy during the apoptosis of TM4 induced by ZEA, and the apoptosis rate of TM4 induced by ZEA was significantly increased. The apoptosis rate of TM4 cells induced by ZEA was significantly decreased. In conclusion ： au-tophagy delayed the apoptosis of TM4 cells and played a protective role in the toxicity of ZEA on TM4 cells.
作者
司梦雪
王冰洁
冯楠楠
邹辉
顾建红
袁燕
刘学忠
刘宗平
卞建春
SI Mengxue;WANG Bingjie;FENG Nannan;ZOU Hui;GU Jianhong;YUAN Yan;LIU Xuezhong;LIU Zongping;BIAN Jianchun(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
《畜牧与兽医》
北大核心
2018年第8期104-109,共6页
Animal Husbandry & Veterinary Medicine
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)
