自由基吸收类抗氧化剂对延边黄牛体细胞克隆重构胚发育的影响

摘要

为提高延边黄牛体细胞克隆重构胚发育率,研究了在体外培养液中单独添加不同浓度的自由基吸收类抗氧化剂维生素E(VE)、表没食子儿茶素没食子酸酯(EGCG)和亚硒酸钠(SS)对克隆重构胚发育率的影响.试验中选择成熟培养20～22 h的卵母细胞,脱颗粒细胞后选择排出第一极体的卵母细胞进行去核与注核操作,然后将融合、激活后的重构胚在添加不同种类不同浓度抗氧化剂的体外培养液中进行培养.结果表明,在卵裂率上,VE的100 μmol·L-1组显著高于其他3组(84.58％ vs 57.64％、70.87％、67.64％)；50和200 μmol·L-1组之间无差异,但都显著高于0 μmol·L-1组(70.87％、67.64％ vs 57.64％).EGCG的7.5和15 μmol·L-1组显著高于加0 μmol· L-1组(79.47％、81.67％ vs 69.47％)且其他2组间差异不显著.SS的添加,5 ng·mL-1组与0和10 ng·mL-1组有显著差异(80.44％ vs 70.27％、67.16％)；与2.5 ng·mL-1组差异不显著(80.44％ vs 73.92％),且其他2组间差异不显著；在囊胚发育率上,VE的100 μmol·L-1组要显著高于其他3组(26.36％ vs 13.04％、18.08％、17.26％).EGCG的15 μmol·L 1组显著高于0和30 μmol·L-1组(24.86％ vs 8.99％、12.10％),和7.5 μmol·L-1组无差异.SS的5与0组和10 ng·mL 1组有显著差异(23.31％ vs 10.32％、12.65％)；2.5 ng·mL 1组与0和10 ng·mL-1l组有显著差异(19.97％ vs 10.32％、12.65％)且其他2组间差异不显著.体外培养液中分别单独添加浓度为100 μmol·L-1、15 μmol·L-1和5 ng·mL-1的维生素E,EGCG和亚硒酸钠都能够显著提高延边黄体细胞克隆牛重构胚的发育率.%In order to improve the development efficiency of somatic cell nuclear transfer embryos of Yanbian Yellow cattle, free radical absorptive antioxidants, such as vitamin E, EGCG and sodium selenite, were added in culture medium in vitro with different concentrations for observing the development of the reconstructed embroys. Oocytes were cultured 20-22 h in maturation medium, oocytes with first polar body were chose for enucleation, injection, integration and activation, then restructured embryos were cultured in in vitro culture mediums with antioxidants with different concentrations. On the cleavage rate, in VE study, results showed that 100 μmol o L-1 group was significantly higher than those of other concertrations(84. 58% vs 57. 64%, 70. 87%, 67. 64%); there ' were no significant differences between 50 and 200 μmol o L-1 groups, but they were both significantly higher than that of 0 μmol o L-1 group(70. 87%, 67. 64% vs 57. 64%); In EGCG study, results showed that 7. 5 and 15 μmol o L-1 groups were significantly higher than that of 0 μmol o L -1 group(79. 47%, 81. 67% vs 69. 47%)and there were no significant differences hetween other groups;In SS study,5 and 0 ng o mL-1 group were significantly higher than 10 ng o ml/-1 group (80.44% vs 70. 27%,67. 16%) while there were no significant differences with 2. 5 ng o mL-1 group(80. 44% vs 73. 92%) , and there were no significant differences between other groups. On the Blastocyst rate,in VE study,results showed that 100 μmol o L-1 group was significantly higher than those of other concertrations(26. 36% vs 13. 04%,18. 08%,17. 26%);In EGCG study, results showed that 15 μmol o L-1 group was significantly higher than 0 and 30 μmol o L-1 groups (24. 86% vs 8. 99%, 12. 10%)and there was no significant differences with 7. 5 μmol o L-1 group;In SS study, 5 ng o mL-1 group was significantly higher than that of 0 and 10 ng o mL-1 groups (23. 31% vs 10. 32%,12. 65%);2. 5 ng o mL-1 group was significantly higher than that of 0 and 10 ng o mL-1 groups(19. 97% vs 10. 32%, 12. 65%) while there were no significant differences between other two groups. Adding 100 μmol o L-1 vitamin E,15 μmol o L-1 EGCG or 5 ng o mL-1 sodium selenite alone in vitro into culture medium can enhance the early development rate of Yanbian Yellow cattle restructured embryos in vitro.