表儿茶素对小鼠卵母细胞mtDNA拷贝数和胚胎体外发育能力的影响

doi:10.16431/j.cnki.1671-7236.2015.04.029

›› 2015, Vol. 42 ›› Issue (4): 966-972.doi: 10.16431/j.cnki.1671-7236.2015.04.029

表儿茶素对小鼠卵母细胞mtDNA拷贝数和胚胎体外发育能力的影响

吴柱连, 刘晓华, 杜凤娇, 雷钏, 尹娜, 郭振伟, 屈春凤, 刘晓淋, 阮秋燕, 陆凤花, 石德顺

广西大学亚热带农业生物资源保护与利用国家重点实验室, 南宁 530005

收稿日期:2014-09-09 出版日期:2015-04-20 发布日期:2015-05-05
通讯作者: 陆凤花, 石德顺 E-mail:lfhgggg@163.com;ardsshi@gxu.edu.cn
作者简介:吴柱连(1988-),女,广西梧州人,硕士生,研究方向:动物繁殖新技术与动物胚胎工程,E-mail:804224345@qq.com;刘晓华(1987-),男,山东临沂人,博士生,研究方向:动物繁殖新技术与动物胚胎工程,E-mail:364306228@qq.com
基金资助:
国家高技术研究发展计划(863)项目(2011AA100607);国家自然科学基金项目(31160457);广西科学基金项目(2012GXNSFFA060004、2011GXNSFA018084);教育部高等学校博士学科点博导基金项目(20114501110001)

Effect of Epicatechin on Mitochondrial DNA (mtDNA) Copy Number and Developmental Competence of Rat Oocytes Following Maturation and Culture in vitro

WU Zhu-lian, LIU Xiao-hua, DU Feng-jiao, LEI Chuan, YIN Na, GUO Zhen-wei, QU Chun-feng, LIU Xiao-lin, RUAN Qiu-yan, LU Feng-hua, SHI De-shun

State Key Laboratory of Subtropical Agricultural Biological Resources Protection and Utilization in Guangxi University, Nanning 530005, China

Received:2014-09-09 Online:2015-04-20 Published:2015-05-05

摘要/Abstract

摘要： 本研究旨在探究表儿茶素(epicatechin,EC)对小鼠体外成熟培养卵母细胞线粒体DNA(mtDNA)拷贝数及其随后孤雌激活胚胎发育能力的影响.小鼠卵丘-卵母细胞复合体(COCs)在添加不同浓度EC(0、5、10、15、20 μmol/L)的成熟液中体外成熟培养16 h后,采用实时荧光定量PCR的方法检测卵母细胞mtDNA拷贝数;同时,通过对卵母细胞进行孤雌激活处理,探讨其后续胚胎的体外发育能力.实时荧光定量PCR分析结果显示,添加 EC各处理组的卵母细胞mtDNA拷贝数均有所增加,其中,10、15 μmol/L组的mtDNA拷贝数均显著高于0 μmol/L对照组(P<0.05);但10 μmol/L组mtDNA拷贝数更接近自然排卵周期合子的mtDNA含量(P>0.05).体外培养观察结果发现,成熟液中添加10 μmol/L EC能提高卵母细胞第一极体排出率,与对照组相比差异不显著(P>0.05),但能显著提高卵母细胞孤雌激活后胚胎的囊胚发育率(P<0.05).综上表明,小鼠卵母细胞体外成熟液中添加10 μmol/L EC可提高卵母细胞的mtDNA拷贝数,有利于促进卵母细胞后续的发育能力.

关键词: 表儿茶素; 卵母细胞; mtDNA; 发育能力

Abstract: This study was designed to investigate the effect of epicatechin (EC) on mitochondrial DNA (mtDNA) copy number and developmental competence of rat oocytes following maturation and culture in vitro. Cumulus-oocyte complexes were respectively cultured in the maturation medium supplemented with different concentrations of EC (0, 5, 10, 15 and 20 μmol/L) for 16 h. And then the mtDNA copy number in matured oocytes was detected with the method of Real-time PCR. Moreover, the subsequent developmental competence of embryos from parthenogenetic activation (PA) was examined. The PCR result showed that the mtDNA copy number in each EC treatment group were increased than that of control group, and there was remarkably increase in the both groups of 10 and 15 μmol/L compared with the control group (P<0.05), but the number of 10 μmol/L group was more closed to that of the nature ovulation cycle oocytes (P>0.05). Furthermore, the result of in vitro culture showed that adding with 10 μmol/L EC into the maturation medium, there were no significant effect in the maturation rate of oocytes (P>0.05), but the developmental rate of blastocyst after PA was significantly improved than that of control group (P<0.05). The datas above showed that adding 10 μmol/L EC into the maturation medium could increase the mtDNA copy number of oocytes, and contribute to improve its subsequent developmental competence.

Key words: epicatechin; oocyte; mtDNA; developmental competence

中图分类号:

S814.9

吴柱连, 刘晓华, 杜凤娇, 雷钏, 尹娜, 郭振伟, 屈春凤, 刘晓淋, 阮秋燕, 陆凤花, 石德顺. 表儿茶素对小鼠卵母细胞mtDNA拷贝数和胚胎体外发育能力的影响[J]. , 2015, 42(4): 966-972.

WU Zhu-lian, LIU Xiao-hua, DU Feng-jiao, LEI Chuan, YIN Na, GUO Zhen-wei, QU Chun-feng, LIU Xiao-lin, RUAN Qiu-yan, LU Feng-hua, SHI De-shun. Effect of Epicatechin on Mitochondrial DNA (mtDNA) Copy Number and Developmental Competence of Rat Oocytes Following Maturation and Culture in vitro[J]. , 2015, 42(4): 966-972.

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表儿茶素对小鼠卵母细胞mtDNA拷贝数和胚胎体外发育能力的影响

Effect of Epicatechin on Mitochondrial DNA (mtDNA) Copy Number and Developmental Competence of Rat Oocytes Following Maturation and Culture in vitro

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