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:2015-03-23
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:2015-03-23

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。

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.

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.

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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

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