EGF对牦牛冻精运动性能、细胞凋亡和IVF胚胎发育潜力的影响  

The Effects of EGF on Yak Frozen Sperm Motility,Apoptosis and the Development Competence of Embryos after IVF

作　　者：潘阳阳[1] 王萌[1] 余四九[1] 崔燕[1] 何俊峰[1] 

机构地区：[1]甘肃农业大学动物医学院甘肃省牛羊胚胎工程技术研究中心,兰州730070

出　　处：《畜牧兽医学报》2017年第5期854-862,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：甘肃省农业生物技术研究与应用开发项目(GNSW-2015-12)

摘　　要：本研究旨在探明牦牛冻精体外获能过程中,EGF是否调控精子的细胞凋亡、运动性能及其对后续胚胎发育的影响。在牦牛冻精体外获能过程中,加入不同浓度EGF(0、50、75、100、150和200ng·mL-1),应用精子分析仪分析不同处理组精子活性;运用qRT-RCR、Western blot检测Bax和Bcl-2的表达;评估IVF后受精胚的发育能力。结果表明:1)EGF可显著提高牦牛冻精运动性能、胚胎卵裂率(P<0.05),且这种调控具有一定的浓度依赖性。2)75~150ng·mL^(-1) EGF可以显著提高获能冻精Bcl-2基因和蛋白表达,降低Bax表达(P<0.05)。综上表明,牦牛冻精体外获能过程中,EGF不仅降低牦牛冷冻精子的细胞凋亡和提高牦牛精子运动性能,而且能够提高IVF后胚胎发育潜力,且其最佳作用浓度为75~150ng·mL^(-1)。本研究为通过生长因子作用提高牦牛体外胚胎生产提供重要参考信息。The study was to explore the effects of epidermal growth factor （EGF） on the apoptosis and motility of yak frozen sperm, the development competence of embryos after in vitro fertilization （IVF）. Frozen-thawed spermatozoa of yak was incubated at 37.5 ℃ for 1 h in Tyrode＇s bicarbonate-buffered medium with EGF at different concentrations （0, 50, 75, 100, 150 and 200 ng·mL-1）. Sperm motility was measured by a computer-assisted sperm analyzer system; the expression of Bax and Bcl-2 in sperm were examined by qRT-PCR and Western blot; the development competence of embryo was also evaluated by using cleavage and blastocyst ratio after IVF. Our results showed that： 1） EGF dramatically enhanced spermatozoa motility and fertilized egg cleavage ratio （P〈0.05）, which were dose-dependence with EGF; 2） The Bcl-2 level was enhanced significantly with EGF at 75-150 ng·mL-1 （P〈0.05）, while the Bax was reduced （P〈0.05）. In conclusion, EGF inhibited the apoptosis and increased the spermatozoa motility of yak frozen sperm during in vitro capacitation, but also enhanced the development competence of embryo after IVF, of which the optimal concentration was 75-150 ng·mL-1. The present study was contributed to improve the yak embryos production in vitro referring to the growth factors.

关 键 词：EGF 牦牛 精子 胚胎 细胞凋亡 

分 类 号：S823.8[农业科学—畜牧学]