母体糖尿病环境对体外受精胚胎早期发育的影响

doi:10.3969/j.issn.1674-5817.2015.02.014

实验动物与比较医学 ›› 2015, Vol. 35 ›› Issue (2): 155-160.DOI: 10.3969/j.issn.1674-5817.2015.02.014

• 华东地区第十三届实验动物科学学术交流会优秀论文选刊 • 上一篇下一篇

母体糖尿病环境对体外受精胚胎早期发育的影响

强苏静¹, 陶凌云², 刘丽均¹, 徐平^1,3, 高诚²

1.上海斯莱克实验动物有限责任公司,上海 201615;
2.上海实验动物研究中心,上海 201203;
3.中国科学院上海生命科学院,上海 201615

收稿日期:2014-06-30 出版日期:2015-04-25 发布日期:2015-04-25
作者简介:强苏静(1987-), 女, 硕士研究生, 研究方向: 动物学。 liyang_sujing@163.com
基金资助:
上海市科委“创新行动计划”重点项目(10140900700); 国家科技支撑计划课题(2013BAK11B02)

Effect of Maternal Diabetic Environment on Development of Early Mouse Embryo by in vitro Fertilization

QIAANG Su jing¹, TAO Ling-yun², LIU Li-jun¹, XU Ping^1,3, GAO Cheng²

1. Shanghai SLAC Laboratory Animal Ltd. Co., Shanghai 201615 China;
2. Shanghai Laboratory Animal Research Center, Shanghai 201203, China;
3. Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, Shanghai 201615, China

Received:2014-06-30 Online:2015-04-25 Published:2015-04-25

摘要/Abstract

摘要： 目的通过腹腔注射链脲佐菌素(streptozotocin, STZ)构建小鼠糖尿病(diabetes mellitus,DM)模型后, 运用体外受精技术研究母体高血糖环境对早期胚胎发育的影响。方法选取6~8周龄ICR雌鼠经2次腹腔注射小剂量STZ构建小鼠糖尿病模型,并设实验对照组。实验母鼠体外受精得到2-细胞胚胎,统计受精率等,并将2-细胞胚胎移植至代孕母鼠体内。取14 d胎龄的胚胎提取总RNA,用Agilent 2100 Bioanalyzer系统对总RNA进行质检合格后,经过逆转录、标记、杂交、洗脱,芯片结果用Affymetrix Scanner 3000进行扫描,用Command Console Software 3.1读取原始数据,实验重复3次,比较实验组与对照组的基因表达的差异情况。结果较高血糖组(血糖为9.5~15.5 mmol/L)母鼠卵子体外受精后发育到2-细胞胚胎的比例为52.3%与对照组的72.1%存在显著统计学差异(P<0.01),高血糖组(血糖值>15.5 mmol/L)胚胎发育到2-细胞的比例为44.2%与对照组相比也存在显著统计学差异(P<0.01)。芯片筛选出差异表达基因121个(P<0.05),其中有119个基因实验组的表达量是对照组的0.5倍以下,2个基因实验组的表达量是对照组的2倍以上。结论糖尿病小鼠的卵子与正常精子体外受精后发育到2-细胞的比例较对照组小鼠低,并且血糖值越高比例越低。由此说明母体糖尿病环境能影响其卵子的生长发育,并且这些影响可能是通过上调代谢相关基因和下调发育相关基因产生的。

关键词: 链脲佐菌素(STZ), 体外受精, 基因芯片, 差异表达

Abstract: Objective The experimental mice were induced to be diabetes mellitus (DM) mice through streptozotocin (STZ) intraperitoneal injection, in order to study the effect of maternal hyperglycemia on early mouse embryo development by in vitro fertilization. Methods ICR mice aged 6~8 weeks were intraperitoneally injected with STZ to established DM models, and the normal mice were considered as control group . The model mice were superovulated by intraperitoneal injection with PMSG and hCG, then the oocytes were fertilized with sperms from normal male mice in vitro. The rates of fertilization were counted and 2-cell embryos were transferred to pseudopregnant mice. Finally 14- dold embryos were obtained to extract. total RNA. After detected by Agilent 2100 Bioanalyzer system, total RNA were reversed transcription, labeled, hybridized, eluted and the data of microarray was scanned by Affymetrix Scanner 3000. Basic data were read by Command Console Software 3.1. The experiments were repeated three times, then the expression of genes between treated group and control group were compared. Results The ratio of two-cell embryo in hyperglycemia group (52.3%), was significantly lower than that in the control group (77.2%) (P<0.01). A total of 121 differentially expressed genes (P<0.05) were screened out. One hundred and nineteen genes expression levels of the experimental group was less 0.5 times than those of the control group, while 2 genes showed a more than 2-fold up regulation. Conclusion After oocytes of model mice fertilizing with sperms of normal male mice in HTF, the rate of embryo developing to two-cell was lower than normal mice. The higher of blood glucose level, the lower of two-cell rate. The growth and development of the oocytes were affected by maternal diabetes environment through up-regulating the metabolism-related genes and down-regulated development-relating genes .

Key words: Streptozotocin(STZ), In vitro fertilization, Microarray, Differentially expression

中图分类号:

Q95-33

强苏静, 陶凌云, 刘丽均, 徐平, 高诚. 母体糖尿病环境对体外受精胚胎早期发育的影响[J]. 实验动物与比较医学, 2015, 35(2): 155-160.

QIAANG Su jing, TAO Ling-yun, LIU Li-jun, XU Ping, GAO Cheng. Effect of Maternal Diabetic Environment on Development of Early Mouse Embryo by in vitro Fertilization[J]. Laboratory Animal and Comparative Medicine, 2015, 35(2): 155-160.

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母体糖尿病环境对体外受精胚胎早期发育的影响

Effect of Maternal Diabetic Environment on Development of Early Mouse Embryo by in vitro Fertilization

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