[1]冯楠楠,王冰洁,朱启航,等.玉米赤霉烯酮通过PI3K/Akt/mTOR通路诱导大鼠睾丸支持细胞自噬及对细胞周期分布的影响[J].南京农业大学学报,2018,41(4):708-714.[doi:10.7685/jnau.201712033]
 FENG Nannan,WANG Bingjie,ZHU Qihang,et al.Effects of zearalenone on the autophagy and cell cycle of sertoli cells by PI3K/Akt/mTOR pathway[J].Journal of Nanjing Agricultural University,2018,41(4):708-714.[doi:10.7685/jnau.201712033]
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玉米赤霉烯酮通过PI3K/Akt/mTOR通路诱导大鼠睾丸支持细胞自噬及对细胞周期分布的影响()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
41卷
期数:
2018年4期
页码:
708-714
栏目:
出版日期:
2018-07-09

文章信息/Info

Title:
Effects of zearalenone on the autophagy and cell cycle of sertoli cells by PI3K/Akt/mTOR pathway
作者:
冯楠楠12 王冰洁1 朱启航1 郑王龙1 邹辉1 顾建红1 袁燕1 刘学忠1 刘宗平12 卞建春12
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
Author(s):
FENG Nannan12 WANG Bingjie1 ZHU Qihang1 ZHENG Wanglong1 ZOU Hui1 GU Jianhong1 YUAN Yan1 LIU Xuezhong1 LIU Zongping12 BIAN Jianchun12
1. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;
2. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China
关键词:
玉米赤霉烯酮睾丸支持细胞G2/M期阻滞PI3K/Akt/mTOR通路自噬
Keywords:
zearalenone(ZEA)sertoli cells(SC)G2/M arrestPI3K/Akt/mTOR pathwayautophagy
分类号:
S851.4
DOI:
10.7685/jnau.201712033
摘要:
[目的]探究磷脂酰肌醇3激酶/蛋白激酶/哺乳动物雷帕霉素靶蛋白(PI3K/Akt/mTOR)信号通路在玉米赤霉烯酮(ZEA)诱导睾丸支持细胞(SC)自噬及影响SC细胞周期分布中的作用,揭示ZEA对雄性生殖毒性的机制。[方法]以Wistar大鼠原代睾丸支持细胞为材料,利用Western blot、流式细胞术和免疫荧光技术等方法检测PI3K/Akt/mTOR信号通路在ZEA对SC自噬及细胞周期分布的影响。[结果]随着ZEA浓度升高,PI3K/Akt/mTOR信号通路关键蛋白磷酸化水平显著或极显著下降(P<0.05或P<0.01);加入PI3K抑制剂LY294002后,微管相关蛋白1轻链3(LC3)的荧光信号强度显著增强,荧光聚点更加明显(P<0.01);同时,与ZEA单独处理组比,Akt、mTOR磷酸化水平显著或极显著下降(P<0.05或P<0.01),LC3Ⅱ/LC3Ⅰ表达量极显著上升(P<0.01);G2/M期关键蛋白Cdc2、Cdc25B等表达水平随着ZEA浓度的升高显著或极显著下降(P<0.05或P<0.01),磷酸化组蛋白H3(p-histone H3)的荧光明显减少;加入PI3K抑制剂LY294002后,ZEA致睾丸支持细胞G2/M期阻滞的比例显著降低(P<0.05),且细胞周期关键激酶Cdc2的活性显著增加(P<0.05)。[结论]通过PI3K/Akt/mTOR信号通路,一定浓度范围的ZEA在诱导SC自噬中起负调控作用,而此通路的激活可以部分逆转ZEA对SC细胞G2/M期的阻滞,从而起到保护作用。
Abstract:
[Objectives]This study aimed to reveal the mechanism of the male reproductive toxicity of zearalenone(ZEA),this study explored the role of PI3K/Akt/mTOR signaling pathway in ZEA induced autophagy in sertoli cells(SC)and its influence on the distribution of cell cycle.[Methods]Using primary SC as test materials,the effects of PI3K/Akt/mTOR signaling pathway on autophagy and cell cycle distribution of SC were detected by Western blot technique,flow cytometry and immunofluorescence techniques.[Results]With the increasing concentration of ZEA,the phosphorylation level of key protein in PI3K/Akt/mTOR pathway decreased gradually(P<0.01 or P<0.05). After adding the PI3K inhibitor LY294002,the fluorescence signal intensity of LC3 significantly increased,which resulted in the more obvious fluorescence accumulation(P<0.01). At the same time,comparing the combined treatment group to the ZEA individual treatment group,the phosphorylation levels of Akt proteins and mTOR proteins significantly decreased(P<0.01 or P<0.05).The expression of LC3Ⅱ/LC3Ⅰ significantly increased(P<0.01). In addition,the expression of key proteins Cdc2,CDC25B in G2/M phase decreased gradually with the increase of ZEA concentrations(P<0.05 or P<0.01). Fluorescence reduction of p-histone H3 was detected by immunofluorescence. After adding the PI3K inhibitor LY294002,the proportion of ZEA induced SC G2/M arrest significantly declined(P<0.05). Moreover,the activity of the cyclic key protease Cdc2 significantly increased(P<0.05).[Conclusions]The results suggest that in a certain concentration range,ZEA can induce autophagy through the activation of the PI3K/Akt/mTOR signaling pathway which plays a negative regulatory role in autophagy process. ZEA can affect the cycle distribution of SC,induce the cell cycle arrest and inhibit proliferation. However,the activation of the PI3K/Akt/mTOR signaling pathway may partially reverse cell G2/M arrest,which plays a protective role.

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备注/Memo

备注/Memo:
收稿日期:2017-12-25。
基金项目: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:冯楠楠,硕士研究生。
通信作者:卞建春,博士,教授,主要从事动物营养代谢病与中毒病研究,E-mail:jcbian@yzu.edu.cn
更新日期/Last Update: 1900-01-01