线粒体凋亡途径在玉米赤霉烯酮致大鼠睾丸支持细胞凋亡中的作用

doi:10.16431/j.cnki.1671-7236.2019.05.031

《中国畜牧兽医》 ›› 2019, Vol. 46 ›› Issue (5): 1516-1524.doi: 10.16431/j.cnki.1671-7236.2019.05.031

线粒体凋亡途径在玉米赤霉烯酮致大鼠睾丸支持细胞凋亡中的作用

朱启航^1,2, 司梦雪¹, 李茜¹, 刘双双¹, 邹辉¹, 顾建红¹, 袁燕¹, 刘学忠¹, 刘宗平^1,2, 卞建春^1,2

1. 扬州大学兽医学院, 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, 扬州 225009

收稿日期:2018-09-26 出版日期:2019-05-20 发布日期:2019-05-20
通讯作者: 卞建春 E-mail:jcbian@yzu.edu.cn
作者简介:朱启航(1994-),女,江苏盐城人,硕士生,研究方向:动物营养代谢病与中毒病,E-mail:904437772@qq.com
基金资助:
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）

Effect of Mitochondria-dependent Pathway on Apoptosis Induced by Zearalenone in Rat Sertoli Cells

ZHU Qihang^1,2, SI Mengxue¹, LI Xi¹, LIU Shuangshuang¹, ZOU Hui¹, GU Jianhong¹, YUAN Yan¹, LIU Xuezhong¹, LIU Zongping^1,2, BIAN Jianchun^1,2

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

Received:2018-09-26 Online:2019-05-20 Published:2019-05-20

摘要/Abstract

摘要：

为探讨玉米赤霉烯酮（zearalenon，ZEA）雄性生殖毒性作用机理，研究ZEA诱导大鼠睾丸支持细胞（sertoli cell，SC）的凋亡途径，试验选取大鼠原代SC为研究材料，用不同浓度ZEA （0、5、10、20 μmol/L）处理SC 24 h后，采用实时荧光定量PCR法检测Bax和Bcl-2 mRNA表达水平；用Western blotting法检测Bax、Bcl-2等线粒体凋亡途径相关蛋白表达；同时检测了20 μmol/L ZEA联合Caspase-8抑制剂（Z-IETD-FMK）处理对SC凋亡率和线粒体凋亡相关蛋白的影响。结果显示，与对照组相比，20 μmol/L ZEA组细胞Bcl-2转录水平显著下降（P<0.05），Bax转录水平显著上升（P<0.05）；10、20 μmol/L ZEA组Bax/Bcl-2比值及Cyto-CytC、Cleaved Caspase-9、Cleaved Caspase-3表达量极显著上升（P<0.01），而Mito-CytC表达量极显著下降（P<0.01）；与ZEA组相比，ZEA与Z-IETD-FMK联合处理可使ZEA诱导的SC凋亡率极显著下降（P<0.01），tBid/Bid、Bax/Bcl-2、Cyto-CytC、Cleaved-Caspase3、Cleaved-Caspase9蛋白表达显著或极显著下降（P<0.05；P<0.01），而Mito-CytC表达量极显著上升（P<0.01）。综合试验结果，ZEA能够通过线粒体途径诱导大鼠SC发生凋亡。

关键词: 玉米赤霉烯酮(ZEA); 睾丸支持细胞(SC); 线粒体凋亡途径; Caspase-8抑制剂

Abstract:

In order to elucidate the mechanism of zearalenone (ZEA) on male reproductive toxicity and investigate the pathway of sertoli cell (SC) apoptosis induced by ZEA,the primary SC was used as the experiment material,exposed to different concentration of ZEA(0,5,10,20 μmol/L) for 24 h.qRT-PCR was used to detect the level of gene transcription of Bax and Bcl-2;Western blotting method was used to detect the expression of proteins related to mitochondria-dependent apoptotic pathway.The changes of apoptosis rate and apoptosis-related protein were detected by 20 μmol/L ZEA combined with the inhibitor of Caspase-8 (Z-IETD-FMK).The results showed that compared with the control group,the level of gene transcription of Bcl-2 was decreased significantly in the group treated with 20 μmol/L of ZEA (P<0.05),while the level of gene transcription of Bax increased significantly (P<0.05).The ratio of Bax/Bcl-2 and the expression of Cyto-CytC,Cleaved caspase-3 and Cleaved caspase-9 were extremely significantly increased while the expression of Mito-CytC was decreased extremely significantly in the group treated with 10,20 μmol/L of ZEA (P<0.01).Compared with ZEA group,the apoptosis rate induced by ZEA decreased extremely significantly after being treated with ZEA and Z-IETD-FMK (P<0.01).The ratio of tBid/Bid and Bax/Bcl-2,Cyto-CytC,Cleaved Caspase-3 and Cleaved Caspase-9 were decreased significantly or extremely significantly (P<0.05;P<0.01),while the Mito-CytC protein expression was increased extremely significantly (P<0.01).The results showed that ZEA could induce the apoptosis of SC via the mitochondria-dependent apoptotic pathway.

Key words: zearalenone(ZEA); sertoli cell(SC); mitochondria-dependent apoptotic pathway; inhibitor of Caspase-8

中图分类号:

Q255

朱启航, 司梦雪, 李茜, 刘双双, 邹辉, 顾建红, 袁燕, 刘学忠, 刘宗平, 卞建春. 线粒体凋亡途径在玉米赤霉烯酮致大鼠睾丸支持细胞凋亡中的作用[J]. 《中国畜牧兽医》, 2019, 46(5): 1516-1524.

ZHU Qihang, SI Mengxue, LI Xi, LIU Shuangshuang, ZOU Hui, GU Jianhong, YUAN Yan, LIU Xuezhong, LIU Zongping, BIAN Jianchun. Effect of Mitochondria-dependent Pathway on Apoptosis Induced by Zearalenone in Rat Sertoli Cells[J]. , 2019, 46(5): 1516-1524.

参考文献

[1] CALDWELL R W,TUITE J,STOB M,et al.Zearalenone production by Fusarium species[J].Appllied Microbiology,1970,20(1):31-34.
[2] HIDY P H,BALDWIN R S,GREASHAM R L,et al.Zearalenone and some derivatives:Production and biological activities[J]. Advances in Applied Microbiology,1977,22:59.
[3] YAZAR S,OMURTAG G Z.Fumonisins,trichothecenes and zearalenone in cereals[J].International Journal of Molecular Sciences,2008,9(11):2062-2090.
[4] FAZEKAS B,TAR A.Determination of zearalenone content in cereals and feedstuffs by immunoaffinity column coupled with liquid chromatography[J].Journal of AOAC International,2001,84(5):1453-1459.
[5] IQBAL S Z,RABBANI T,ASI M R,et al.Assessment of aflatoxins,ochratoxin A and zearalenone in breakfast cereals[J].Food Chemistry,2014,157(33):257-262.
[6] GRENIER B,APPLEGATE T J.Modulation of intestinal functions following mycotoxin ingestion:Meta-analysis of published experiments in animals[J].Toxins,2013,5(2):396-430.
[7] KIM I H,SON H Y,CHO S W,et al.Zearalenone induces male germ cell apoptosis in rats[J].Toxicology Letters,2003,138(3):185-192.
[8] 肖治军.猪玉米赤霉烯酮中毒与繁殖障碍[J].中国畜牧兽医,2005,32(2):45-46. XIAO Z J.Porcine zearalenone poisoning and reproductive disorders[J].China Animal Husbandry & Veterinary Medicine,2005,32(2):45-46.(in Chinese)
[9] YANG J,ZHANG Y,WANG Y,et al.Toxic effects of zearalenone and alpha-zearalenol on the regulation of steroidogenesis and testosterone production in mouse Leydig cells[J].Toxicology in Vitro,2007,21(4):558-565.
[10] SYED V,HECHT N B.Disruption of germ cell-sertoli cell interactions leads to spermatogenic defects[J].Molecular & Cellular Endocrinology,2002,186(2):155-157.
[11] RD W W,KARSH L,GITTES R F.The role of germinal epithelium and spermatogenesis in the privileged survival of intratesticular grafts[J].Journal of Urology,1985,134(4):782-786.
[12] CCAMERON D F,WHITTINGTIO K,SCHULTZ R E,et al.Successful islet/abdominal testis transplantation does not require Leydig cells[J].Transplantation,1990,50(4):649-653.
[13] ZHENG W,PAN S,WANG G,et al.Zearalenone impairs the male reproductive system functions via inducing structural and functional alterations of sertoli cells[J].Environmental Toxicology & Pharmacology,2016,44(3):146-155.
[14] XU M L,HU J,GUO B P,et al.Exploration of intrinsic and extrinsic apoptotic pathways in zearalenone-treated rat sertoli cells[J].Environmental Toxicology,2016,31(12):1731-1739.
[15] KANTAR C,WALCZAK H.Caspase-8 and Bid:Caught in the act between death receptors and mitochondria[J].Biochimica et Biophysica Acta-Molecular Cell Research,2011,1813(4):558-563.
[16] GUO B,ZHAI D,CABEZAS E,et al.Humanin peptide suppresses apoptosis by interfering with Bax activation[J].Nature,2003,423(6938):456-461.
[17] LIU X,KIM C N,POHL J,et al.Purification and characterization of an interleukin-1 beta-converting enzyme family protease that activates cysteine protease P32(CPP32)[J].Journal of Biological Chemistry,1996,271(23):13371-13376.
[18] 王卫东,陈正堂.Bcl-2/Bax比率与细胞"命运"[J].中国肿瘤生物治疗杂志,2007,14(4):393-396. WANG W D,CHEN Z T.Bcl-2/Bax ratio and the "life or death fate" of cells[J].Chinese Journal of Cancer Biotherapy,2007,14(4):393-396.(in Chinese)
[19] 李环,赵金昌,徐一波,等.DBP对大鼠睾丸支持细胞内Ca²⁺、线粒体膜电位及Caspase活性的影响[J].北华大学学报(自然科学版),2015,16(4):454-457. LI H,ZHAO J C,XU Y B,et al.Effects of DBP on intracellular Ca²⁺,mitochondrial membrane potential and Caspase activity in sertoli cells in rats[J].Journal of Beihua University (Natural Science),2015,16(4):454-457.(in Chinese)
[20] 张振,王为,董煜,等.二硫化碳对大鼠睾丸支持-生精共培养细胞凋亡线粒体途径相关基因表达的影响[J].环境与职业医学,2013,30(11):870-874. ZHANG Z,WANG W,DONG Y,et al.Effect of carbon disulfide on expression of genes related with mitochondrion mediated sertoli-germ cells apoptosis in male rats[J].Journal of Environmental & Occupational Medicne,2013,30(11):870-874.(in Chinese)
[21] LI H,ZHU H,XU C J,et al.Cleavage of BID by caspase 8 mediates the mitochondrial damage in the Fas pathway of apoptosis[J].Cell,1998,94(4):491-501.
[22] KUWANA T,SMITH J J,MUZIO M,et al.Apoptosis induction by caspase-8 is amplified through the mitochondrial release of cytochrome C[J].Journal of Biological Chemistry,1998,273(26):16589-16594.
[23] LI M,KONDO T,ZHAO Q L,et al.Apoptosis induced by cadmium in human lymphoma U937 cells through Ca²⁺-calpain and caspase-mitochondria-dependent pathways[J].Journal of Biological Chemistry,2000,275(50):39702-39709.
[24] LEE J,LEE S,KIM S L,et al.Corn silk maysin induces apoptotic cell death in PC-3 prostate cancer cells via mitochondria-dependent pathway[J]. Life Sciences,2014,119(1-2):47-55.

线粒体凋亡途径在玉米赤霉烯酮致大鼠睾丸支持细胞凋亡中的作用

Effect of Mitochondria-dependent Pathway on Apoptosis Induced by Zearalenone in Rat Sertoli Cells

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