分泌型卷曲相关蛋白家族成员在多囊卵巢患者不同成熟度卵母细胞周围的卵丘颗粒细胞中的基因表达

目的探讨在多囊卵巢综合征(PCOS)患者与非PCOS患者中,分泌型卷曲相关蛋白(sFRPs)家族成员基因在不同成熟度卵母细胞周围的卵丘颗粒细胞上的表达差异。方法收集27例接受体外受精-胚胎移植(IVF-ET)助孕治疗的不育女性患者(其中11例PCOS患者,16例非PCOS患者),通过控制性促排卵治疗后,采用标准长方案促排卵,经阴道超声引导下穿刺取卵后,获得共198枚卵丘-卵母细胞复合体(PCOS患者98枚,非PCOS患者100枚);采用机械剥离法获取卵丘颗粒细胞,根据卵母细胞成熟度不同,将对应的卵丘颗粒细胞分为4组:非PCOS患者成熟卵母细胞周围的颗粒细胞组(CCN-MII),非PCOS患者非成熟卵母细胞周围的颗粒细胞组(CCN-MI+GV),PCOS患者成熟卵母细胞周围的颗粒细胞组(CCP-MII)和PCOS患者非成熟卵母细胞周围的颗粒细胞组(CCP-MI+GV)。通过实时荧光定量聚合酶链反应(qRT-PCR)技术检测部分sFRPs家族基因成员(sFRP1、sFRP2、sFRP3、sFRP4、sFRP5)在PCOS及非PCOS患者不同成熟度卵母细胞周围的卵丘颗粒细胞上的表达情况。结果 (1)PCOS患者中,在未成熟卵母细胞周围的颗粒细胞中sFRP4的表达显著高于在成熟卵母细胞周围的颗粒细胞中的表达,即sFRP4在CCP-MⅠ+GV组表达高于CCP-MⅡ组[(7.80±1.21)vs.(1.00±0.00)](P0.05);(2)在非PCOS患者中,sFRPs家族成员sFRP4和sFRP5在不同成熟度卵泡母细胞周围的卵丘颗粒细胞中表达变化显著,CCN-MⅠ+GV组中的表达量均显著高于CCN-MⅡ组[sFRP4(7.04±1.30)vs.(1.00±0.00)](P0.05)和[sFRP5(2.84±0.73)vs.(1.00±0.00)](P0.05);(3)sFRP4的表达在PCOS及非PCOS患者间成熟卵母细胞对应的颗粒细胞组间均无显著性差异。结论在PCOS患者中,仅sFRP4在卵丘颗粒细胞上的表达与卵母细胞的成熟度相关;而在非PCOS患者中,sFRP4和sFRP5的表达均与卵母细胞的成熟度相关,其表达量随卵母细胞成熟而降低。

Expression of sFRPs family members in cumulus cells around oocytes at different mature stage in patients with/without polycystic ovary syndrome

Objective： To investigate gene expression profile of sFRP family members in cumulus ceils around oocytes at different mature stages in patients with/without polycystic ovary syndrome（PCOS）. Methods： After controlled ovarian stimulation（COS）, a total of 198 cumulus cell （CC） samples （CCpcos = 98, CCnon PCOS = 100） were obtained from 27 patients （PCOS = 11, non-PCOS = 16 ） during IVF cycles. All the CC samples were divided into four groups：CCN-MI1, CCN MI＋GV, CCp-MI1, CCp-MI＋GV based on the maturation of oocyte. Quantitative real time-PCR（qRT-PCR）was performed in this study. Results： sFRP4 expression in CCpMI ＋cv was significantly higher than that in CCp-MⅡ [（7.80±1.21）vs. （1.00±0.00）]（P〈0.05）in PCOS patients. In the non-PCOS patients,both of sFRP4[（7.04±1.30）vs. （1.00±0.00）land sFRPS[（2.84±0. 73） vs. （1.00±0.00）]expressions in CCN-Mi ＋GV were significantly higher than those in CCN-MⅡ in（P〈0.05）. The expression of sFRP4 had no significant difference in cumulus cells around oocytes at same mature stages between PCOS and non-PCOS patients. Conclusions： These observations indicated that sFRP5 expression was decreased along with oocyte maturation in both PCOS and non-PCOS patients. However, sFRP4 expression is only associated with oocyte maturation in PCOS patients.