单卵母细胞对应的卵丘颗粒细胞中生长分化因子-9和骨形成蛋白-15的表达水平与胚胎发育潜能的研究  

Expression of GDF9 and BMP15 mRNA level in cumulus cells of single oocyte and its correlate with oocyte development potency in human

作　　者：徐清华[1] 程立立 张敏[1] 包莉莉[1] 许鹏宇 李冬秀[1] XU Qing-hua;CHENG Li-li;ZHANG Min;BAO Li-li;XU Peng-yu;LI Dong-xiu(Bethune lnternational Heping Hospital Reproductive Center,Shifiazhuang,05008)

机构地区：[1]白求恩国际和平医院生殖中心,石家庄050082

出　　处：《中国优生与遗传杂志》2018年第9期116-120,共5页Chinese Journal of Birth Health & Heredity

基　　金：国家卫生计生委计划生育与优生重点实验室(项目编号:2015003)

摘　　要：目的评估单卵母细胞对应的卵丘颗粒细胞中GDF9和BMP15的m RNA表达水平对卵母细胞发育潜能的影响。方法采用长方案治疗25例ICSI助孕的女性,取卵当天收集180份单卵母细胞对应的卵丘颗粒细胞,随后提取卵丘颗粒细胞的m RNA,进行实时荧光定量分析,对GDF9和BMP15 m RNA基因表达水平采用独立样本t检验分析。结果常规长方案CCs 180份,因卵子与收集的卵丘颗粒细胞是一一对应的,根据卵子受精情况将CCs分为正常受精组CCs 129份,异常受精组CCs 51份;正常受精组根据第3日胚胎卵裂情况分为优质卵裂胚形成组CCs 58份,非优质卵裂胚形成组CCs71份;非优质卵裂胚继续培养,根据第5日形成囊胚情况,分为优质囊胚形成组CCs 25份,非优质囊胚形成组CCs 46份。GDF9和BMP15 m RNA基因表达水平与正常受精率,优质胚胎率显著相关。正常受精组GDF9和BMP15 m RNA基因表达水平显著高于非正常受精组(P<0.01),优质胚胎组显著高于非优质胚胎组(P<0.01)。结论单卵母细胞对应的卵丘颗粒细胞中GDF9和BMP15 m RNA基因表达水平与正常受精率,优质胚胎率和优质囊胚形成率呈显著正相关,提示其可以作为预测卵母细胞的发育潜能的指标。Objective：The aims of the present study were to detect the expression levels of GDF9 and BMP15 mRNAs in cumulus cells of sing oocyte and to analyze the correlation between their expression levels and the oocyte quality and developmental potential. Methods：A total of 180 individual cumulus cells of single oocyte from 25 women were undergo the treatment of l CSI. The TRizol Reagent was used to extract total RNA from each CCs sample according to the instructions,relative mRNA expression levels of the studied genes were quantified using real-time PCR analysis. To compare the differences between the levels of cytokines for independent t-test,the difference was statistically significant. Results：The CCs were 180. Because oocytes and cumulus cells is one-to-one,according to oocyte fertilization,embryo development and blastocyst formation,retrospectively divided the CCs into different groups. ⅰ）The CCs were divided into two groups of normal fertilization group（CCs=129）and abnormal fertilization group（CCs=51）according to fertilization case.ⅱ）Normal fertilization group were divided into two groups of good quality embryos（CCs=58）and poor quality embryo（CCs=71）on 3 day according to embryos development. ⅲ）poor quality embryos were divided into two groups of good quality blastocyst（CCs=25）and poor quality blastocyst（CCs=46）development according to blastocyst development on 5 day. Through this experiment can be seen, the expression level of GDF-9 and BMP15 gene mRNA of normal fertilization group were significantly higher than the abnormal fertilization group（P〈0.01）,good quality embryos group higher than poor quality embryos（P〈0.01）,and good quality blastocyst group higher than poor quality embryos（P〈0.01）. Conclusion： This study demonstrated that expression levels of GDF-9 and BMP-15 in CCs are positive correlation with the oocyte quality and developmental potential. It is revealed that human cumulus cells in GDF-9 and BMP-15 level could predict the development pote

关 键 词：生长分化因子-9 骨形成蛋白-15 卵子成熟 胚胎质量 

分 类 号：R321-33[医药卫生—人体解剖和组织胚胎学]