延边黄牛体细胞克隆胚胎氧化损伤的初步研究

为了优化体细胞克隆胚胎体外培养液,提高体细胞的克隆效率,以CRlaa+5% FBS+0.3%BSA为基础培养液,分别添加0、60、80、100和120 μmol/L H_2O_2观察延边黄牛体细胞克隆胚胎的氧化损伤情况和体外发育模式,并研究1、4和7 mmol/L GSH对延边黄牛重组胚的保护作用.结果表明:(1)添加0、60、80 μmol/LH_2O_2组卵裂率显著要高于100 μmol/L和120 μmol/L组(P<0.05),添加H_2O_2的组囊胚发育率显著低于未添加组(P<0.05);(2)随着H_2O_2处珲浓度的升高,重组胚发育速度增加,但是发育停滞现象也更明显;(3)在培养液中添加GSH后,1 mmol/L组卵裂率显著高于7 mmol/L组(P<0.05),与0和4 mmol/L组差异不显著(P>0.05),囊胚发育率显著高于其它组(P<0.05).可见,在体细胞克隆胚胎体外发育过程中H_2O_2对其具有氧化损伤作用,不利其体外发育,在培养液中添加1 mmol/L GSH对体细胞克隆胚胎的氧化损伤具有明显的保护作用.

A Study on Oxidative Damage to Reconstructed Embryos from Somatic Cell Nuclear Transfer in Yanbian Cattle

In order to optimize the culture solution and improve the efficiency of somatic cell nuclear transfer,oxidative damage and developmental mode in vitro of reconstructed embryos from somatic cell nuclear transfer in Yanbian catde were observed in the experiment treated with addition of H_2O_2 to the base culture so-lution(CRlaa+5%FBS+0.3%BSA),and the protective effect of GSH on reconstructed embryos was studied.The results showed that:(1)The cleavage rate of the group with addition of 0,60 and 80 μmol/L of H_2O_2 was significantly higher than that of 100 and 120 μmol/L(P<0.05).The blastoeyst rate of the group treated with H_2O_2 was significantly lower than that of the group without the treatment of H_2O_2(P<0.05);(2)The rate of development of reconstructed embryos increased along with the rise of the concentration of H_2O_2,but the arrest phenomenon of development was much more obvious;(3)The cleavage rate of the group with addition of 1 mmol/L of GSH was significantly higher than that of the group treated with 7 mmol/L(P<0.05),but was not significantly hither than that of the group treated with 0 and 4 mmol/L(P>0.05).The blastocyst rate of the group treated with 1 mmol/L was strikingly higher than that of the other treatment groups(P<0.05).It is thus evident that oxidative damage by H_2O_2 exists in the development of reconstructed embryos from so-matic cell nuclear transfer,and it is harmful for the development of reconstructed embryos in vitro.An addition of 1 mmol/L of GSH in culture solution possesses obvious protective effect on oxidative damage to reconstructed embryos from somatic cell nuclear transfer.