卵母细胞分泌因子在人卵丘颗粒细胞中的表达及其与卵母细胞发育潜能的关系  

Expression of oocyte-secreted factors in human cumulus cells and its relationship with oocyte developmental potential

作　　者：吴小华 程立立 张敏[1] 焦保权[1] 朱爱萍[1] 常秀峰[1] 徐清华[1] 李冬秀[1] 包莉莉[1] 许鹏宇[1] 

机构地区：[1]白求恩国际和平医院妇产科生殖医学中心,石家庄050082

出　　处：《生殖与避孕》2016年第9期701-707,共7页Reproduction and Contraception

基　　金：2013年全军后勤科研项目;项目编号:BBJ13C001

摘　　要：目的:探讨单卵母细胞所对应的卵丘颗粒细胞(cumulus cells,CCs)中生长分化因子-9(GDF-9)、骨形成蛋白-15(BMP-15)、转化生长因子β1(TGFβ1)、Smad3的表达与对应单卵母细胞质量及其发育潜能的关系以及在不同促排卵方案中的表达。方法:因男方因素或曾有过体外受精(IVF)不受精史接受卵胞质内单精子显微注射(ICSI)治疗的患者41例,单卵母细胞的CCs 332份(Gn RH-a组21例172份CCs,非垂体降调节组20例160份CCs)。拣卵后机械法剥离单卵子周围的CCs,将卵母细胞与收集的CCs一一对应进行分析。M_Ⅱ卵均采用ICSI授精,并观察其受精、卵裂及囊胚发育情况。采用Real-time PCR方法检测,比较各组CCs中GDF-9、BMP-15、TGFβ1和Smad3的相对含量。结果:1 CCs收集及PCR扩增产物可满足实验要求。2 Gn RH-a组及非垂体降调节组中,上述4种卵母细胞分泌因子,在正常受精组的相对含量均显著高于异常受精组;优质卵裂胚形成组的相对含量均显著高于非优质卵裂胚形成组;优质囊胚形成组中的相对含量显著高于非优质囊胚形成组。3 Gn RH-a组的CCs中BMP-15的表达量相对更高;而GDF-9、TGFβ1和Smad3在非降调节组的CCs中相对表达量较高。结论:1采用单卵母细胞的CCs的RNA进行研究,实际操作是可行的,RNA的质量和数量可达到实验要求。2 GDF-9、BMP-15、TGFβ1、Smad3可作为评价卵母细胞质量、预测卵母细胞发育潜能的客观指标。3降调节与否可能影响CCs中的卵母细胞分泌因子的表达水平,且与M_Ⅱ期卵母细胞发育潜能有关。Objective： To explore the prognostic value of oocyte-secreted factors （OSFs） for the quality and developmental potential of oocytes, expression of growth differentiation factor 9 （GDF-9）, bone morphogenetic protein-15 （BMP-15）, transforming growth factor-β1 （TGFI31） and Smad3 from individual cumulus cells （CCs） were analyzed corresponding to each cumulus cell-oocyte complex （COC） in two protocols that pituitary down-regulation and non-down-regulation. Methods： The individual CC masses of total 332 COCs from 41 women undergoing in vitro fertilization （IVF） were collected. The infertile women were determined to undergo intracytoplasmic sperm injection （ICSI） because of the male factor or with a history of failed/low fertilization at previous IVF. All CCs from each COCs were mechanically removed shortly after oocyte retrieval, and all M~ oocytes were subject to ICSI, and subsequengtly assessed on the basis of their fertilization, cleavage, and blastocyst development. Total RNA was extracted from each CCs sample according to the manufacturers＇ protocols. RNA expression of GDF-9, BMP-15, TGFβ1 and Smad3 was assessed using a step Real-time quantitative PCR assay. Results： 1） The quality and quantity of RNA extracted from CCs could meet the PCR experiment requirements. 2） In total individual CCs, the expression levels of GDF-9, BMP-15, TGFβ1 and Smad3 were significantly higher in normal fertilization group than in abnormal fertilization group, were significantly higher in good-quality cleavage embryo group than in poor-quality cleavage embryo group, and were significantly higher in good-quality blastocyst group than in poor-quality blastocyst group. 3） The expression levels of BMP-15 were significantly increased in GnRH-a protocol. However, the expression levels of GDF-9, TGFβ1 and Smad3 in non-pituitary down-regulation were significantly higher compared with GnRH-a protocol. Conclusion： 1） Gene expression data were available on CCs from all 332 individuals of COC, a

关 键 词：卵母细胞分泌因子(OSFs) 卵丘颗粒细胞(CCs) 卵胞质内单精子显微注射(ICSI) 胚胎发育潜能 卵母细胞质量 GnRH-a长方案 非降调节促排卵方案 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]