生长激素改善多囊卵巢综合症患者卵母细胞成熟度的研究

Growth hormone improves oocyte maturity in patients with polycystic ovary syndrome

目的 探讨生长激素(GH)在控制性超排卵(COH)过程中改善PCOS患者卵母细胞成熟度的应用价值及其初步机制。 方法 选择2016年9月至2019年1月在深圳市妇幼保健院生殖医学中心利用卵胞浆内单精子注射(ICSI)技术进行不孕治疗的PCOS患者75例。根据患者促排卵过程中是否使用外源GH将其分成两组,使用GH组(44例)和未使用GH组(31例)。取卵后离体卵母细胞经体外培养3~4h后,对成熟的MⅡ卵行ICSI授精,余下的未成熟卵继续体外培养16h,培养成熟的MⅡ卵再行ICSI授精。比较两组患者继续培养前后卵母细胞成熟情况及其MⅡ卵授精与胚胎培养结局。检测两组患者血液与卵泡液GH、E2及IGF-I含量的差异,以及卵巢颗粒细胞上相应受体基因表达的不同,并分析卵泡液GH含量与其它指标的相关性。 结果 离体卵母细胞经体外培养3~4h后,与未使用GH组相比,使用GH组患者MⅡ卵数和总MⅡ卵率均显著升高(P<0.05),而MⅠ卵数和GV卵数显著减少(P<0.05)。ICSI授精后,使用GH组患者的正常受精率、卵裂率和优胚率均显著高于未使用GH组(P<0.05),而周期取消率显著低于后者(P<0.05)。授精后余下的未成熟卵继续培养16h后,两组间各指标的差异均无统计学意义(P>0.05)。进一步比较,发现使用GH组患者血清和卵泡液GH与IGF-1含量及卵泡液E2浓度和卵巢颗粒细胞上GHRmRNA、IGF-1RmRNA和StARmRNA的表达水平均显著高于未使用GH组(P<0.05),且卵泡液中GH含量与卵泡液IGF-1和E2水平、MⅡ卵数以及颗粒细胞上GHRmRNA、IGF-1RmRNA和StARmRNA相对表达量均显著正相关(P<0.05)。 结论 超排卵过程中添加外源GH对改善PCOS患者的卵母细胞成熟度具有临床应用价值。其改善卵母细胞成熟度的作用可能依赖于配体-受体途径及IGF-1介导的途径所建立的激素和细胞因子微环境。

Objective : To explore the application value and preliminary mechanism of exogenous growth hormone(GH)in improving oocyte maturity in patients with polycystic ovary syndrome(PCOS)during controlled ovarian hyperstimulation(COH). Methods : Seventy five patients with PCOS undergoing ICSI-ET for infertility treatment in the Department of Reproduction Center of Shenzhen Maternal & Child Healthcare Hospital from September 2016 to January 2019 were selected.The patients were divided into two groups according to whether exogenous GH was used during ovulation induction:GH group( n =44)and non-GH group( n =31).Isolated oocytes were cultured in vitro for 3-4 hours after oocyte retrieval,and metaphase II oocytes were fertilized by ICSI.Then the remaining immature oocytes were cultured in vitro for another 16 hours,and the cultured mature metaphase II stage oocytes in vitro were also fertilized by ICSI.The oocyte maturation before and after continuous culture and the outcome of MII oocyte fertilization and embryo culture were compared between the two groups.The GH,E 2 and IGF-1 levels in blood and follicular fluid,as well as the expression of the corresponding receptor genes of the ovarian granulosa cells were detected and compared between the two groups.The correlations between the GH level in follicular fluid and other indicators were analyzed. Results : After that isolated oocytes were cultured in vitro for 3-4 hours,the number and rate of metaphase II stage oocytes in GH group were significantly higher than those in non-GH group( P <0.05),while metaphaseⅠstage oocytes and GV oocytes were significantly decreased( P <0.05).After fertilizing by ICSI,the normal fertilization rate,the cleavage rate and the good quality embryo rate in GH group were significantly higher than those in non-GH group ( P <0.05),but the cycle cancellation rate was significantly lower ( P <0.05).The remained immature oocytes after fertilization were cultured continuously for 16 hours,all the indexes were no significantly different between the two groups( P >0.05).In addition,the concentration of GH and IGF-1 in serum and follicular fluid,the level of E 2 in follicular fluid and the expression levels of GHR mRNA,IGF-1R mRNA and StAR mRNA of ovarian granulosa cells in GH group were significantly higher than those in non-GH group( P <0.05).The concentration of GH in follicular fluid was significant positively correlated with the levels of IGF-1 and E 2 ,number of metaphase II stage oocytes,expression levels of GHR mRNA,IGF-1R mRNA and StAR mRNA of ovarian granulosa cells( P <0.05). Conclusions : Adding exogenous GH during COH has clinical application value in improving oocyte maturity of PCOS patients.Its effect on improving oocyte maturation may depend on the hormone and cytokine microenvironment established by ligand-receptor pathway and IGF-1 mediated pathway.