SAHA对水牛卵母细胞体外成熟及胚胎发育潜能的影响  

Effect of SAHA on Development Potency of Oocytes and Embryos of Buffalo(Bubalus bubalus)

作　　者：陆杏蓉 梁贤威[1] 邓廷贤[1] 朱鹏[1] 段安琴 庞春英[1] 

机构地区：[1]中国农业科学院广西水牛研究所/广西水牛遗传繁育重点实验室,南宁530001

出　　处：《农业生物技术学报》2016年第9期1450-1456,共7页Journal of Agricultural Biotechnology

基　　金：广西科技项目(桂渔牧科201452032和桂科合14123001-5);广西水牛研究所基本科研业务费(水牛基1504004)

摘　　要：卵母细胞体外成熟过程中组蛋白修饰发生了剧烈的变化,其表达模式的改变对卵母细胞的成熟乃至后续胚胎的发育有着至关重要的作用。去乙酰化酶抑制剂-伏立诺他(suberoylanilide hydroxamic acid,SAHA)作为新型抗癌药物,是组蛋白去乙酰化酶的潜在抑制剂,在体内和体外抑制多种类型肿瘤细胞生长并促进其分化和凋亡。为探讨组蛋白去SAHA对水牛(Bubalus bubalus)卵母细胞体外成熟和胚胎体外发育潜能的影响,本研究比较了不同浓度(0,2,5,10,20和40 nmol/L)SAHA处理成熟期的水牛卵母细胞对其成熟率(第一极体排出率)、受精卵分裂率、囊胚率和囊胚总细胞数的影响。结果表明,10 nmol/L SAHA处理组成熟率、卵裂率、囊胚率显著高于对照组((77.07±1.79)%和(62.3±2.08)%,(86.81±1.93)%和(74.86±2.07)%,(27.56±2.86)%和(24.23±1.74)%,P<0.05)。随着浓度增加,各处理组囊胚总细胞数有提高的趋势,但组间差异不显著(141±10,151±13,165±17,170±14,147±20,185±22,P>0.05)。q RT-PCR检测结果显示,SAHA对水牛卵母细胞成熟过程的处理,均显著下调了c AMP反应元件结合蛋白(c AMP responsive element binding protein,CREB)结合蛋白基因CBP、组蛋白乙酰转移酶1基因(histone acetyltransferases 1,HAT1)、组蛋白去乙酰化酶1基因(histone deacetylase 1,HDAC1)在MⅡ期卵母细胞中的表达,P300则有所上调。实验结果表明,组蛋白去乙酰化酶抑制剂SAHA处理是一种能够提高水牛卵母细胞体外成熟效率促进胚胎发育的有效途径之一,该研究结果可为今后研究转基因克隆胚胎发育相关基因调控机理提供了理论基础。There is dramatic changing of histone modifications during in vitro maturating of oocytes, the changing of its expression pattern plays an important role on oocyte maturation and subsequently embryonic development. Suberoyl anilide hydroxamic acid （SAHA） acting as a anticancer drug is the potential inhibitor of histone deacetyltransferase, inhibiting the growth, differentiation and apoptosis of various types of tumor cell in vivo and vitro. To explore the effect of inhibitor of histone deacetyltransferase SAHA in vitro maturation and embryonic development potential of buffalo （Bubalus bubalus） oocytes, effects of maturing rate （first polar body discharge rate）, cleavage rate of fertilized eggs, blastocyst rate and total cell number of blastocyst of buffalo oocytes were compared by different concentrations of （0, 2, 5, 10, 20 and 40 nmol/L） SAHA treatment. Results showed that maturing rate, cleavage rate, and blastocyst rate of group 10 nmol/LSAHA was significantly higher than control group （（77.07±1.79）% and （62.3±2.08）%, （86.81±1.93）% and （74.86±2.07）%, （27.56±2.86）% and （24.23±1.74）%, P〈0.05）. As the concentration increasing, total cell number of blastocyst in each treatment group had a tendency to increase, but had no significantly difference with each other （141±10, 151±13, 165±17, 170±14, 147±20 and 185±22, P〉0.05）. qRT-PCR tests revealed that the expression of （cAMP responsive element binding protein, CREB） binding protein gene （CBP）, histone acetyltransferases 1 （HAT1） gene and histonedeacetylase I（HDAC1） gene in M II stage oocytes of treatment groups were significantly reduced, though p300 was increased. In conclusion, histone deacetylase inhibitors SAHA was one of the most effective way to promote maturation of buffalo oocytes and embryonic development in vitro, this result will provid a theoretical basis for future study about gene regulatory mechanism of transgenic cloned embrvonic-developmental.

关 键 词：水牛 卵母细胞 胚胎发育 SAHA 组蛋白去乙酰化 

分 类 号：S823.83[农业科学—畜牧学]