摘要
目的研究一氧化氮(NO)在小鼠囊胚发育和孵化中的调控作用和机制。方法从妊娠第2.5天雌鼠子宫角获取桑葚胚进行体外培养48 h。胚胎培养液中分别添加不同浓度N-硝基-L-精氨酸甲酯(L-NAME)或硝普钠(SNP),以及L-NAME与SNP联合添加。分别于第4天和第5天统计囊胚发育率和孵化率,激光扫描共聚焦显微镜观察统计囊胚总细胞数和活化型含半胱氨酸的天冬氨酸蛋白水解酶3(caspase 3)表达。结果随着L-NAME和SNP浓度增加,囊胚发育率、孵化率和囊胚总细胞数显著降低。在5 mmol/L L-NAME培养液中添加10 nmol/L SNP可显著提高囊胚发育率和孵化率;但随着SNP浓度增加,囊胚发育率和孵化率显著降低。L-NAME对囊胚细胞中活化型caspase 3表达没有显著影响,而500 nmol/L以上浓度SNP可显著提高囊胚细胞中活化型caspase 3表达,促进囊胚细胞凋亡。结论 NO在小鼠囊胚发育和孵化中可发挥重要的调控作用,囊胚细胞中NO浓度过低或过高都将会影响卵裂球分裂,降低囊胚发育率和孵化率,且过高浓度NO会引起囊胚细胞凋亡。
Objective To determine the regulatory role and mechanism of nitric oxide（ NO） in the development and hatching of mouse blastocysts. Methods The Kunming female mice were superovulated and then mated with mature male mice. On the day 2. 5 of their pregnancy,morulae were flushed from their uterine horns with culture media. Morulae were cultured in different concentrations of N-nitro-L arginine methyl ester（ L-NAME）,sodium nitroprusside（ SNP）,or the combination of L-NAME and SNP in culture media for 48 hours. The development and hatching of blastocysts were examined on day 4 and day 5 and the total numbers of blastocyst cells and cysteinyl aspartate specific proteinase 3（ caspase 3） were observed under confocal laser scanning microscope.Results With the increase of the concentration of L-NAME or SNP,the hatching rate of blastocysts and the total number of blastocyst cells were significantly reduced. The addition of 10 nmol / L SNP in culture media with5 mmol / L L-NAME significantly increased the development of blastocysts and promoted hatching of blastocysts.However,with increase of SNP concentration in culture media with 5 mmol / L L-NAME,the development and hatching rates of blastocysts were significantly decreased. L-NAME had no obvious effect on the expression of active caspase 3 in blastocyst cells. However,when being above 500 nmol / L,SNP significantly increased the expression of caspase 3 in blastocyst cells. Conclusions NO plays an important role in development and hatching of mouse blastocysts. Excessively high or low NO can damage the division of blastomeres,resulting in the failure of the blastocyst development and hatching. Also,excessively high NO can lead to the apoptosis of the blastocyst cells.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2015年第5期501-507,共7页
Acta Academiae Medicinae Sinicae
基金
吉林省教育厅"十二五"科学技术研究项目[吉教科合字(2013)第349号]
山东省自然科学基金(ZR2011HL005)
吉林省科技发展计划项目[201201077]
吉林医药学院大学生科研基金[吉医学科字(2012)第04号]~~
关键词
小鼠
一氧化氮
囊胚发育
囊胚孵化
凋亡
mouse
nitric oxide
blastocyst development
blastocyst hathing
apoptosis
