卵母细胞分泌因子在卵巢储备功能减退患者中的表达及对胚胎发育的影响

Expression of oocyte secreted factor and its relationship with embryonic development in patients with diminished ovarian reserve

目的分析卵母细胞分泌因子在卵巢储备功能减退(DOR)患者卵丘颗粒细胞(CCs)中的表达情况,预测不同的促排卵方案对卵母细胞成熟及胚胎发育的影响,为DOR不孕女性寻找更有效的促排卵方案。方法收集接受卵胞浆内单精子显微注射(ICSI)助孕治疗的80例DOR患者,425份CCs为研究对象,其中微刺激方案组30例171份CCs,促性腺激素释放激素拮抗剂(GnRH-A)方案组30例165份CCs,PPOS方案组20例89份CCs,单卵丘颗粒细胞群与卵母细胞一一对应。提取卵母细胞周围CCs的mRNA,采用实时荧光定量聚合酶链反应检测各组CCs中生长分化因子-9(GDF-9)、骨形成蛋白-15(BMP-15)和转化生长因子-β1(TGF-β1)相对表达量。结果GnRH-A方案组中采用促性腺激素(Gn)天数及Gn剂量明显多于其余两组(P<0.05)。在3种促排卵方案中,MⅡ期GDF-9、BMP-15和TGF-β1相对表达量高于MⅠ和GV期;正常受精组的GDF-9、BMP-15和TGF-β1相对表达量高于异常受精组;优质胚胎组的GDF-9、BMP-15和TGF-β1相对表达量高于非优质胚胎组(P<0.05);优质囊胚组的GDF-9、BMP-15和TGF-β1相对表达量明显高于非优质囊胚组(P<0.05)。在卵母细胞阶段,GnRH-A方案组GDF-9、BMP-15相对表达量明显高于微刺激方案组、PPOS方案组(P<0.05);微刺激方案组BMP-15相对表达量明显低于PPOS方案组(P<0.05);其余各组间GDF-9、BMP-15、TGF-β1相对表达量比较,差异无统计学意义(P>0.05)。结论GnRH-A方案可能增加采用Gn天数及Gn剂量,GDF-9、BMP-15和TGF-β1可作为评估DOR患者卵母细胞成熟度及其发育潜能的客观指标,对于高龄DOR患者,微刺激方案、GnRH-A方案和PPOS方案均可选择。

Objective To analyze the expression of oocyte secretion factor in cumulus granulosa cells(CCs)of patients with diminished ovarian reserve(DOR),to predict the influence of different ovulation induction schemes on oocyte maturation and embryo development,and to find more effective ovulation induction schemes for infertile women with DOR.Methods A total of 425 CCs samples were collected from 80 DOR patients who received Intracytoplasmic Sperm Injection(ICSI)as pregnancy aid treatment,including 171 CCs samples from 30 patients treated with microstimulation(microstimulation group),165 CCs samples from 30 patients treated with antagonist(GnRH-A group),and 89 CCs samples from 20 patients treated with PPOS(PPOS group).The monozygotic granulosa cell populations were one-to-one corresponding to oocytes.The mRNA of CCs around oocytes was extracted.The relative expression levels of growth differentiation factor-9(GDF-9),bone morphogenetic protein-15(BMP-15)and transforming growth factor-β1(TGF-β1)in each group of CCs were detected by real-time fluorescence quantitative polymerase chain reaction,then compared.Results The days and dose of Gn in GnRH-A group were significantly higher than those in other two groups(P<0.05).The relative expression levels of GDF-9,BMP-15 and TGF-β1 in the MⅡstage were higher than those in the MⅠand GV stages.The relative expression levels of GDF-9,BMP-15 and TGF-β1 in normal fertilization group were significant higher than those in abnormal fertilization group(P<0.05).The relative expression levels of GDF-9,BMP-15 and TGF-β1 in high-quality embryo group were significant higher than those in non-high-quality embryo group(P<0.05).The relative expression levels of GDF-9,BMP-15 and TGF-β1 in high quality blastocyst group were significant higher than those in non-high quality blastocyst group(P<0.05).The relative expression levels of GDF-9 and BMP-15 in GnRH-A group were significant higher than those in micro-stimulus group and PPOS group(P<0.05).The relative expression level of BMP-15 in micro stimulation group was significantly lower than that in PPOS group(P<0.05).There was no significant difference on the relative expression levels of GDF-9,BMP-15 and TGF-β1 among other groups(P>0.05).Conclusion GnRH-A regimen may increase the number of days and dose of Gn.GDF-9,BMP-15 and TGF-β1 could be used as objective indicators to evaluate oocyte maturity and developmental potential in patients with DOR.For elderly DOR patients,microstimulation regimen,GnRH-A regimen and PPOS regimen could be selected.