牛磺熊脱氧胆酸对黄牛体外受精早期胚胎发育的影响

doi:10.11843/j.issn.0366-6964.2015.03.007

畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (3): 395-401.doi: 10.11843/j.issn.0366-6964.2015.03.007

牛磺熊脱氧胆酸对黄牛体外受精早期胚胎发育的影响

李政达^#，阮子芸^#，雷钏，阮秋燕，蒋建荣，石德顺^*，陆凤花^*

（广西大学亚热带生物资源保护利用国家重点实验室，南宁 530004)

收稿日期:2014-10-28 出版日期:2015-03-23 发布日期:2015-03-24
通讯作者: 石德顺,研究员，博导,E-mail:ardsshi@gxu.edu.cn；陆凤花,研究员，博导,E-mail:lfhgggg@163.com
作者简介:李政达（1987-），男，壮族，广西南宁人，硕士，主要从事动物胚胎工程研究，E-mail:LZD-OK@163.con；阮子芸（1988-）,女，广西玉林人，博士生，主要从事动物胚胎工程研究，E-mail:ziyunruan@163.com。李政达和阮子芸为并列第一作者
基金资助:
国家农业部转基因重大专项（2011ZX08007-003）；国家高技术研究发展计划（863）项目（2011AA100607）；教育部高等学校博士学科点博导基金项目(20114501110001)；广西八桂学者项目；广西科学基金项目(2012GXNSFFA060004)；广西研究生教育创新计划项目（YCBZ2014012）

Effects of Tauro Ursodeoxychollc Acid (TUDCA) on Development of Bovine IVF Embryos

LI Zheng-da^#， RUAN Zi-yun^#，LEI Chuan，RUAN Qiu-yan，JIANG Jian-rong，SHI De-shun^*，LU Feng-hua^*

（State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources，Guangxi University，Nanning 530004，China）

Received:2014-10-28 Online:2015-03-23 Published:2015-03-24

摘要/Abstract

摘要：

旨在研究牛磺熊脱氧胆酸（Tauro ursodeoxychollc acid，TUDCA）对黄牛体外受精（IVF）早期胚胎发育的影响。采用RT-PCR法检测黄牛IVF胚胎早期发育各时期XBP-1 mRNA剪接激活情况，随后分别用添加不同浓度（0、100、250、500、1 000 μmol•L^-1）TUDCA的培养液培养胚胎，分别统计胚胎分裂率、囊胚率、囊胚细胞总数及囊胚细胞凋亡率，采用qRT-PCR法分析囊胚内质网应激相关基因Grp78、Chop、Bax、Bcl-2的表达情况。结果表明，在胚胎的4-细胞、桑椹胚、囊胚检测到XBP-1 mRNA明显剪接；IVF胚胎经不同浓度TUDCA培养后各组间的胚胎分裂率差异不显著（P>0.05），但500 μmol•L^-1TUDCA组提高了胚胎的囊胚发育率及囊胚细胞数（P<0.05），并降低了囊胚细胞凋亡率（P<0.05）；qRT-PCR检测表明，500 μmol•L^-1TUDCA处理组下调了囊胚Chop基因的表达，且上调了Bcl-2基因的表达（P<0.05），而Grp78与Bax表达与对照组相比差异不显著（P>0.05）。综上表明：早期胚胎体外发育过程中，存在内质网应激反应；胚胎培养液中添加500 μmol•L^-1TUDCA，可以降低内质网应激反应，有利于促进黄牛胚胎发育。

关键词: 黄牛, TUDCA, IVF, 内质网应激, 基因表达

Abstract:

This study was conducted to investigate the effects of tauro ursodeoxychollc acid (TUDCA) on the development of bovine in vitro fertilization (IVF) embryos.Firstly，the splicing of XBP-1 mRNA of the embryos at different developmental stages was detected with RT-PCR.Then，the bovine IVF embryos were cultured in the medium containing different concentration of TUDCA（0，100，250，500，1 000 μmol•L^-1），respectively，and the embryonic cleavage rate，blastocyst rate，the blastocyst cell number as well as apoptosis rate of blastocyst were investigated.Meanwhile，the expression of endoplasmic reticulum stress (ERS) related genes were detected by qRT-PCR.The results showed that：XBP-1 mRNA splicing was observed during embryonic development，especially in 4-cell，morula and blastocyst stages.No significant effect were observed in the cleavage rate of IVF embryos after treating with different concentration of TUDCA，while in the group treated with 500 μmol•L^-1 TUDCA，the blastocyst rate as well as the cell number of blastocyst were increased significantly (P<0.05)，moreover，the cells apoptosis rate of blastocysts in this group was decreased significantly (P<0.05).The results of qRT-PCR showed that 500 μmol•L^-1TUDCA supplemented into the culture medium for bovine IVF embryos could significantly down-regulate the expression level of Chop gene and up-regulate Bcl-2 gene (P<0.05)，however，the expressions of Grp78 and Bax gene were no significant difference as compared to that of control group (P>0.05).These results indicated that ERS was present in the process of bovine embryos in vitro culture(IVC)，and supplementing with 500 μmol•L^-1TUDCA into the embryo culture medium might decrease ERS，and promote embryonic development.

Key words: bovine, TUDCA, IVF, endoplasmic reticulum stress, gene expression

中图分类号:

S823.8⁺1.2

李政达，阮子芸，雷钏，阮秋燕，蒋建荣，石德顺，陆凤花. 牛磺熊脱氧胆酸对黄牛体外受精早期胚胎发育的影响[J]. 畜牧兽医学报, 2015, 46(3): 395-401.

LI Zheng-da， RUAN Zi-yun，LEI Chuan，RUAN Qiu-yan，JIANG Jian-rong，SHI De-shun，LU Feng-hua. Effects of Tauro Ursodeoxychollc Acid (TUDCA) on Development of Bovine IVF Embryos[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(3): 395-401.

参考文献

［1］LANE M,GARDNER D K.Understanding cellular disruptions during early embryo development that perturb viability and fetal development［J］.Reprod Fert Dev，2005，17(3)：371-378.
［2］BELL C E，LARIVI K.UNDERSTANDING C，et al.Mitogen-activated protein kinase(MAPK) pathways mediate embryonic responses to culture medium osmolarity by regulating Aquaporin 3 and 9 expression and localization，as well as embryonic apoptosis［J］.Hum Reprod，2009，24(6)：1373-1386.
［3］PAULA-LOPES F,HANSEN P.Heat shock-induced apoptosis in preimplantation bovine embryos is a developmentally regulated phenomenon［J］.Biol Reprod，2002,66(4):1169-1177.
［4］ABRAHAM T，PIN C L，WATSON A J.Embryo collection induces transient activation of XBP1 arm of the ER stress response while embryo vitrification does not［J］.Mol Hum Reprod, 2012，18(5)：229-242.
［5］LEE K，TIRASOPHON W，SHEN X,et al.IRE1-mediated unconventional mRNA splicing and S2P-mediated ATF6 cleavage merge to regulate XBP1 in signaling the unfolded protein esponse［J］.Genes Dev，2002,16(4)：452-466.
［6］OYADOMARI S,MORI M.Roles of CHOP/GADD153 in endoplasmic reticulum stress［J］.Cell Death & Differ，2004,11(4)：381-389.
［7］MCCULLOUGH K D，MARTINDALE J L，KLOTZ L O，et al.Gadd153 sensitizes cells to endoplasmic reticulum stress by down-regulating Bcl2 and perturbing the cellular redox state［J］.Mol Cell Biol，2001,21(4)：1249-1259.
［8］GORMAN A M，HEALY S J，STO J L,et al.Stress management at the ER：regulators of ER stress-induced apoptosis［J］.Pharm & Therap, 2012,134(3)：306-316.
［9］OMURA T，ASARI M，YAMAMOTO J，et al.Sodium tauroursodeoxycholate prevents paraquat-induced cell death by suppressing endoplasmic reticulum stress responses in human lung epithelial A549 cells［J］.Biochem Bioph Res Co，2013,432(4)：689-694.
［10］LEE Y Y，HONG S H，LEE Y J，et al.Tauroursodeoxycholate (TUDCA)，chemical chaperone，enhances function of islets by reducing ER stress［J］.Biochem Bioph Res Co，2010,397(4)：735-739.
［11］ZHANG J Y，DIAO Y F，KIM H R,et al.Inhibition of endoplasmic reticulum stress improves mouse embryo development［J］.PLoS ONE，2012，7(7)：e40433.
［12］陆凤花.水牛体细胞核移植的研究［D］.南宁：广西大学，2004.
LU F H.The research of somatic cell nuclear transfer of buffalo［D］.The dissertation of Guangxi University,2004.(in Chinese)
［13］SONG B S，KIM J S，YOON S B，et al.Inactivated Sendai-virus-mediated fusion improves early development of cloned bovine embryos by avoiding endoplasmic-reticulum-stress-associated apoptosis［J］. Reprod Fert Dev，2011,23(6)：826-836.
［14］LEE A H，IWAKOSHI N N，GLIMCHER L H.XBP-1 regulates a subset of endoplasmic reticulum resident chaperone genes in the unfolded protein response［J］.Mol Cell Biol，2003,23(21)：7448-7459.
［15］IWAWAKI T，AKAI R，KOHNO K，et al.A transgenic mouse model for monitoring endoplasmic reticulum stress［J］.Nat Med，2003,10(1)：98-102.
［16］SOUID S，LEPESANT J A，YANICOSTAS C.The xbp-1 gene is essential for development in Drosophila［J］.Dev Genes Evol，2007,217(2)：159-167.
［17］ZHANG J Y，DIAO Y F，OQANI R K,et al.Effect of endoplasmic reticulum stress on porcine oocyte maturation and parthenogenetic embryonic development in vitro［J］.Biol Reprod，2012,86(4)：128.
［18］KIM J S，SONG B S，LEE K S,et al.Tauroursodeoxycholic acid enhances the pre-implantation embryo development by reducing apoptosis in pigs［J］. Reprod Domest Anim，2012,47(5)：791-798.
［19］GUPTA M K，UHM S J，HAN D W，et al.Embryo quality and production efficiency of porcine parthenotes is improved by phytohemagglutinin［J］.Mol Reprod Dev, 2007,74(4)：435-444.

[1]	宋慧子, 栾兆进, 王兆琛, 杜炜, 赵勇超, 张家新. 永生化绵羊附睾上皮细胞系的建立及其生物学特性分析[J]. 畜牧兽医学报, 2019, 50(9): 1822-1831.
[2]	李华振, 狄冉, 刘秋月, 胡文萍, 王翔宇, 马琳, 刘武军, 储明星. 类视黄醇X受体基因调控动物季节性发情和发情周期的研究进展[J]. 畜牧兽医学报, 2019, 50(8): 1525-1535.
[3]	李丽敏, 邹云婧, 郝雪飘, 赵兴华, 李睿文, 孙继国, 袁万哲. 脑心肌炎病毒感染神经细胞的基因表达谱分析[J]. 畜牧兽医学报, 2019, 50(8): 1709-1714.
[4]	李华振, 狄冉, 刘秋月, 胡文萍, 王翔宇, 马琳, 刘武军, 储明星. 类视黄醇X受体基因调控动物季节性发情和发情周期的研究进展[J]. 畜牧兽医学报, 2019, 50(8): 1525-1535.
[5]	李丽敏, 邹云婧, 郝雪飘, 赵兴华, 李睿文, 孙继国, 袁万哲. 脑心肌炎病毒感染神经细胞的基因表达谱分析[J]. 畜牧兽医学报, 2019, 50(8): 1709-1714.
[6]	占思远, 丁雪, 仲涛, 王林杰, 李利, 张红平. IGF1R和IGF2R在南江黄羊不同组织和肌细胞中的表达模式比较[J]. 畜牧兽医学报, 2019, 50(4): 701-711.
[7]	刘同君, 赵盼, 赵敏孟, 刘龙, 崔恒宓, 龚道清, 耿拓宇. 内质网应激标记基因Grp78参与鹅肥肝免疫/炎症状态的调控[J]. 畜牧兽医学报, 2019, 50(4): 727-737.
[8]	王波, 刁志成, 马勇, 李发弟, 曲扬华, 徐晨晨, 卢晓楠, 罗海玲. 日粮添加油脂和出生类型对羔羊肌肉胶原蛋白特性及相关基因表达的影响[J]. 畜牧兽医学报, 2019, 50(4): 771-780.
[9]	占思远, 丁雪, 仲涛, 王林杰, 李利, 张红平. IGF1R和IGF2R在南江黄羊不同组织和肌细胞中的表达模式比较[J]. 畜牧兽医学报, 2019, 50(4): 701-711.
[10]	刘同君, 赵盼, 赵敏孟, 刘龙, 崔恒宓, 龚道清, 耿拓宇. 内质网应激标记基因Grp78参与鹅肥肝免疫/炎症状态的调控[J]. 畜牧兽医学报, 2019, 50(4): 727-737.
[11]	王波, 刁志成, 马勇, 李发弟, 曲扬华, 徐晨晨, 卢晓楠, 罗海玲. 日粮添加油脂和出生类型对羔羊肌肉胶原蛋白特性及相关基因表达的影响[J]. 畜牧兽医学报, 2019, 50(4): 771-780.
[12]	王惠影, 郭保地, 何大乾, 龚绍明, 刘毅, 李光全, 徐琪, 陈国宏. 饲粮蛋氨酸对鹅再生羽绒品质、血清指标及基因mRNA表达量的影响[J]. 畜牧兽医学报, 2019, 50(2): 323-331.
[13]	王惠影, 郭保地, 何大乾, 龚绍明, 刘毅, 李光全, 徐琪, 陈国宏. 饲粮蛋氨酸对鹅再生羽绒品质、血清指标及基因mRNA表达量的影响[J]. 畜牧兽医学报, 2019, 50(2): 323-331.
[14]	辛海瑞, 郝月, 张闯, 顾宪红. 葡萄糖调节蛋白94在内质网应激诱导肝细胞凋亡中的调控机制研究进展[J]. 畜牧兽医学报, 2019, 50(10): 1963-1971.
[15]	张秋旭, 张润厚, 史晓雪, 李刚, 高爱琴. 添加不同形式亚麻油对肉羊生产性能、肉品质、脂肪酸含量及脂代谢相关酶基因mRNA表达量的影响[J]. 畜牧兽医学报, 2018, 49(9): 1928-1939.

牛磺熊脱氧胆酸对黄牛体外受精早期胚胎发育的影响

Effects of Tauro Ursodeoxychollc Acid (TUDCA) on Development of Bovine IVF Embryos

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价