摘要
目的运用体外受精技术改变糖尿病小鼠胚胎的早期发育环境，研究母体糖尿病环境对早期胚胎基因表达的影响。方法选取6-8周龄ICR雌鼠2次腹腔注射小剂量STZ构建小鼠糖尿病模型，模型雌鼠超数排卵后通过体外受精的方法将培养至2一细胞的胚胎转移至正常假孕ICR雌鼠的输卵管内，取14d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 Software3．1读取原始数据，实验重复3次。结果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。结论母体糖尿病环境能影响其卵子的生长发育，通过上调代谢相关基因和下调发育相关基因影响早期胚胎的生长发育。
Objective To study the effect of maternal diabetes environment on early fetuses gene expression through in vitro Fertilization（IVF） technology. Methods Diabetes animal model was induced by a low dose of STZ in 6-8 weeks ICR female mice, and then the mice with hyperglycemia were superovulated. Two-cell embryos derived from in vitro fertilization were transferred into the oviduct of normal pseudopregnant ICR female mouse. Total RNA of 14 days＇ fetuses were collected and detected by Agilent 2100 Bioanalyzer system. Through subsequent reversed transcription, label, hybridization, elution and microarray scann by Affymetrix Scanner 3000, finally raw data was read by Command Console Software 3.1. The experiment was repeated three times. Results 121 differentially expressed genes （P〈0.05） were screened out. The expression level of 119 genes in the experimental group was less 0.5 times than that in the control group, and it shown more than 2-fold up regulation. Conclusion Maternal diabetes environment influenced the growth and development of the oocytes through up-regulating the metabolism-related genes and down-regulated development-relating genes and further influenced the growth and development of the early embryos.
出处
《实验动物与比较医学》
CAS
2013年第2期84-89,共6页
Laboratory Animal and Comparative Medicine
基金
上海市科委“创新行动计划”重点项目（10140900700）
关键词
糖尿病小鼠
体外受精
基因芯片
差异表达
Diabetes mouse
In vitro Fertilization（IVF）
Microarray
Differential expression＇
