ERK信号通路对玉米赤霉烯酮诱导大鼠睾丸支持细胞G2期阻滞的影响  

Effects of ERK signaling pathway on G2 phase arrest of rat’s sertoli cells induced by zearalenone

作　　者：包若昱 王冰洁[1] 冯楠楠[1] 邹辉[1] 袁燕[1] 顾建红[1] 刘学忠[1] 刘宗平[1,2] 卞建春 BAO Ruo-yu;WANG Bing-jie;FENG Nan-nan;ZOU Hui;YUAN Yan;GU Jian-hong;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 Disease and Zoonoses,Yangzhou 225009,China)

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《中国兽医科学》2018年第12期1594-1600,共7页Chinese Veterinary Science

基　　金：国家重点研发计划支持项目(2016YFD0501208);江苏省大学生学术科技创新基金项目(201711117090X);江苏高校优势学科建设工程资助项目(PAPD)

摘　　要：为揭示玉米赤霉烯酮（zearalenone,ZEA）雄性生殖毒性的机理,以雄性Wistar大鼠原代睾丸支持细胞（SC）为材料,对ERK信号通路在ZEA对睾丸支持细胞G2期阻滞中的作用进行了研究。采用20 mol/L的ZEA对SC进行染毒,分别采用U0126（ERK抑制剂）共处理、ERK siRNA沉默ERK基因的方法,利用流式细胞术、免疫荧光技术和Western-blot等技术对SC细胞周期的变化及细胞周期相关蛋白进行检测。结果显示,与ZEA组相比,U0126＋ZEA共处理组G2/M期细胞比例极显著降低（P〈0.01）,p53、p21蛋白的表达量极显著下降（P〈0.01）,关键激酶cdc2的活性极显著增强（P〈0.01）,cdc25B的蛋白表达量显著增加（P〈0.05）,CyclinB1表达有所上升但差异不显著（P〉0.05）,表达p-H3的细胞数显著增多。与ZEA染毒组相比,ERK1/2 siRNA＋ZEA组G2/M期细胞比例均极显著降低（P〈0.01）;ERK1/2 siRNA＋ZEA组的cdc2活性均增加（P〈0.05或P〈0.01）,cdc25B的蛋白表达量均升高（P〈0.05或P〈0.01）,CyclinB1的蛋白表达量均极显著升高（P〈0.01）。上述结果表明,ERK信号通路通过调控G2期相关蛋白参与了ZEA诱导的睾丸支持细胞G2期阻滞。In order to investigate the mechanism of male reproductive toxicity of zearalenone（ZEA）,the effects of zearalenone on sertoli cell cycle and ERK signaling pathway in G2 arrest of sertoli cells were studied in male Wistar rats.SC was contaminated with 20 mol/L ZEA.The method of U0126（ERK inhibitor） coprocessing with ZEA,or ERK si RNA silencing were used respectively.The flow cytometry,immunofluorescence technique and Western-blot were used to detect the changes of SC cell cycle and cell cycle related proteins.In result,compared with the ZEA（20 mol/L） treatment group,the proportion of G0/G1 phase cells in the U0126（10 mol/L）＋ZEA treatment group was extremely significant lower than that in G2/M phase cells（P〈0.01）.The expression of the p53 and p21 protein was significantly decreased（P〈0.01）.The activity of the cdc2 was significantly increased（P〈0.01） and the protein expression of cdc25 B was also significantly increased（P〈0.05）.The expression of the Cyclin B1 was slightly increased（P〈0.05）,while the amount of the p-H3 increased obviously.Compared with the ZEA treatment groups,the proportion of G2/M phase cells in the group of the ERK siRNA significantly decreased（P〈0.01）,while the activity of the cdc2 was increased（P〈0.05 or P〈0.01）.The protein expression of cdc25 B was significantly increased（P〈0.05 or P〈0.01）.The protein expression the Cyclin B1 was also extremely increased（P〈0.01）.The results showed that ERK signaling pathway participates in the G2 arrest of the sertoli cells induced by ZEA through the regulation of the G2 related proteins.

关 键 词：玉米赤霉烯酮 睾丸支持细胞 G2/M期阻滞 ERK信号通路 

分 类 号：S852.659.5[农业科学—基础兽医学]