中药有效成分对小鼠2-细胞胚胎体外发育的作用  

Effect of Chinese medicine active ingredients on mouse 2-cell embryos in vitro development

作　　者：高建明[1] 孙玉成[1] 陈武[1] 于同泉[2] 穆祥[1] 杨柳[2] 路萍[2] 

机构地区：[1]北京农学院动物科学技术系,北京102206 [2]北京市农业应用新技术重点实验室,北京102206

出　　处：《广西农业生物科学》2006年第B09期203-204,共2页Journal of Guangxi Agricultural and Biological Science

基　　金：北京市委组织部优秀人才培养专项经费D类项目;北京市农业应用新技术实验室开放课题项目(KF2003-04)

摘　　要：如何获得更多具有活力的可操作的桑椹胚、囊胚一直是胚胎体外培养的研究热点。近二十年来，虽经采用改进培养液成分、改善培养条件、建立体细胞与胚胎共培养体系和添加生长因子、细胞因子等手段，胚胎质量有所提高，但体外受精卵的囊胚发育率仍远低于体内受精，与体内受精囊胚相比，体外受精囊胚的细胞总数和内细胞团细胞数量明显减少，胚胎移植妊娠率和产仔率较低，这些问题均严重影响了胚胎工厂化生产和胚胎产业化的发展。以及育种工作的进程。因此，探索开辟胚胎体外培养生长发育的新途径，提高其发育率和移植妊娠率显得尤为必要。目前，国内外有关中草药有效成分对体外胚胎发育作用的研究甚少。本研究以ICR小鼠2-细胞胚胎为试验材料，将中药有效成分黄芩苷（Baicalin，Bai）、川芎嗪（Ligustrazine，Lig）和小檗碱（Berberine，BR）添加于mCZB基础液中，并以添加双抗的mCZB基础液为试验对照组，比较其对胚胎体外发育的影响。结果表明，体外培养120h的胚胎孵化率，Bai组（80．6％）、Lig组（78．3％）及BR组（75．9％）极显著高于对照组（51．8％）（P〈0．01）。孵化胚胎细胞计数，BR组、Lig组和Bai组（分别为87．2±8．6、83．9±7．7和81．9±6．2），与对照组（77．4±5．6）差异极显著（P〈0．01）。说明3种中药有效成分均能显著促进小鼠早期胚胎的体外发育及胚胎细胞增殖。为进一步探讨Bai、Lig、BR对胚胎体外培养发育的效果和验证其胚胎质量，将不同组别培养至桑椹、囊胚期的胚胎进行两种程序常规冷冻，解冻后继续培养，观察其冷冻效果，并将各组解冻后发育至囊胚期胚胎移植。结果表明：冷冻解冻桑椹胚体外培养8～14h囊胚发育率，程序一中Bai组（65.8％）和Lig组（81．1％）分别显著（P〈0．05）和极显著（P〈0．01The study was compared the effects of Baicalin （Bai）, Ligustrazine （Lig） and Berberine （BR） added to mCZB medium on 2-cell mouse embryos in vitro culture. The results showed that the rates of hatched embryos at 120 h from the three groups of Bai （80. 6%）, Lig （78.3%） and BR （75.9%） was significant higher than that of control group （51.8%）, respectively （P〈0. 01）. The differences of the cell amount of hatched embryos between treated groups （ B1R ： 8 7. 2 ±8. 6 , Lig ： 83. 9±57. 7 and Bai ： 81.9±56.2） and control group （77.4±55.6） were significant （P〈0.01） . Three kinds of Chinese medicine active ingredients could efficiently promote development of mouse early embryos in vitro and increase embryo cells number. The morula and blastocyst in vitro culture were conserved through two freezing process for validating effects of Bai, Lig and BR on embryos quality. The frozen-thawed embryos were cultured to the stage of blastula and transferred to recipient uterus. The results showed that the development rate of morula cultured for 8～14 h after frozen-thawed in the first procedure of the Bai was 65.8% whichwas superior to that of the contro/ （48.6%） and the BR （46.7%） （P〈0. 05）, which of the Lig （81.1%） was significantly higher than that of the control and the BR （P〈0.01）. In the second procedure, the development rates of the Bai （80.6%） and the Lig （78.3%） were significantly higher than that of the control（55.20% ） and the BR（47.6%） （P〈0.01）. The development rates of blastocyst cultured for 6～8 h in the first procedure, there was no significant difference （P〉0.05） between each group . In the second procedure , the effects of treatments were superior to that of control group （P〈0.05） . There was. no significant differences between the different procedures （P〉0.05） . The transfer pregnancy rates and the birth rates from the three groups of Bai （56.3%, 63.3% ）, Lig （52. 6%, 67.3% ）

关 键 词：中药有效成分 2-细胞胚胎 体外培养 冷冻解冻 胚胎移植 小鼠 

分 类 号：S814.8[农业科学—畜牧学]