分泌型卷曲相关蛋白家族成员在多囊卵巢综合征患者卵丘颗粒细胞中的基因表达分析

发布时间：2018-03-28 12:09

  本文选题：sFRPs家族成员　切入点：卵丘颗粒细胞　出处：《青岛大学》2014年硕士论文

【摘要】：目的探讨部分分泌型卷曲相关蛋白(sFRPs)家族成员基因在多囊卵巢综合征(PCOS)患者与非PCOS患者不同成熟度卵母细胞对应的卵丘颗粒细胞上的基因表达差异。 方法收集27例接受体外受精-胚胎移植(IVF-ET)助孕治疗的不孕女性(其中PCOS患者11例,非PCOS患者16例),采用标准长方案促排卵,经阴道超声引导下穿刺取卵后,采用机械法剥离每个卵母细胞的卵丘颗粒细胞,共获得197枚卵丘-卵母细胞复合体(PCOS患者97枚,非PCOS患者100枚),机械剥离每个卵母细胞周围的部分卵丘颗粒细胞,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患者成熟卵母细胞的颗粒细胞组(CCN-XI=MII),非PCOS患者未成熟卵母细胞的颗粒细胞(CCN_N-MI+GV),PCOS患者成熟卵母细胞的颗粒细胞组(CCP_MII)和PCOS患者未成熟卵母细胞的颗粒细胞(CCP-MI+GV)。通过实时荧光定量PCR反应(qRT-PCR)方法技术检测部分sFRPs家族基因成员(sFRPl, sFRP2, sFRP3, sFRP4, sFRP5)在PCOS及非PCOS患者不同成熟度卵母细胞相应的卵丘颗粒细胞上的表达情况。 结果(1)PCOS患者中,sFRP4在未成熟卵母细胞的颗粒细胞中的表达显著高于其在成熟卵母细胞的颗粒细胞中的表达,在CCP-MI+GV组sFRP4表达高于CCP-MⅡ组[(7.80±1.21)vs(1.00±0.00)](P0.05)；(2)sFRPs家族成员中sFRP4和sFRP5在非PCOS患者不同成熟度卵母细胞的卵丘颗粒细胞中表达变化明显,CCN-MI+GV组中的表达量明显都高于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无显著差异。 结论sFRP4在卵丘颗粒细胞上的表达与卵母细胞的成熟度相关,其表达量随着卵母细胞的核成熟而逐渐降低。而且,在非PCOS患者中,sFRP5的这种表达变化亦与卵母细胞的核成熟度相关。
[Abstract]:Objective to investigate the difference of gene expression in cumulus granulosa cells corresponding to different maturation oocytes in polycystic ovary syndrome (PCOS) patients with polycystic ovary syndrome (PCOS) and non PCOS patients. Methods 27 infertile women (11 patients with PCOS and 16 patients with non-#en1#) who received IVF-ET-assisted in vitro fertilization and embryo transfer assisted pregnancy were collected. Ovulation was induced by standard long protocol, and the ovulation was obtained by transvaginal ultrasound guided puncture. Mechanical stripping of cumulus granulosa cells from each oocyte was used to obtain 97 cumulus-oocyte complexes from 97 PCOS patients and 100 non-PCOS patients. Some of the cumulus granulosa cells around each oocyte were removed mechanically. Depending on the maturation of the oocyte, The corresponding cumulus granulosa cells were divided into four groups: the granulosa cells of the mature oocytes from non-#en0# patients, the granulosa cells from the immature oocytes from the non-#en1# patients, the granulosa cells from the mature oocytes from the patients with PCOS and the patients with PCOS. The expression of some members of sFRPs family, sFRPl, sFRP2, sFRP3, sFRP4, sFRP5, in the corresponding cumulus granulosa cells of PCOS and non-#en3# patients with different maturity oocytes was detected by real-time fluorescence quantitative PCR reaction (QRT-PCR). Results the expression of sFRP4 in granulosa cells of immature oocytes was significantly higher than that in granulosa cells of mature oocytes in PCOS patients. The expression of sFRP4 and sFRP5 in the CCN-MI GV group was higher than that in the CCP-M 鈪，

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