解剖学报

解剖学报 ›› 2018, Vol. 49 ›› Issue (2): 233-239.doi: 10.16098/j.issn.0529-1356.2018.02.016

• 组织学胚胎学发育生物学 • 上一篇    下一篇

卵丘颗粒细胞骨形发生蛋白-15、6信号通路分子表达对受精卵发育能力的影响

林典梁1 全松2* 康跃凡1 余爱丽1 林元1   

  1. 1. 福建省妇幼保健院，福建医科大学附属医院，福建省生殖医学中心，福州 350001;2. 南方医科大学南方医院妇产科生殖医学中心，广州 510515
  • 收稿日期:2017-07-24 修回日期:2017-10-10 出版日期:2017-04-06 发布日期:2018-04-06
  • 通讯作者: 全松 E-mail:quansong@smu.edu.cn
  • 基金资助:
    福建省卫生系统中青年骨干人才培养项目

Effect of signaling molecules bone morphogenetic protein-15，6 expression in oophorus cumulus granulosa cells on the developmental ability of human fertilized oocytes

LIN Dian-liang1 QUAN Song 2* KANG Yue-fan1 YU Ai-li1 LIN Yuan1   

  1. 1. Fujian Maternity and Children’s Hospital, Fujian Provincial Reproductive Medicine Center， Affiliated Hospital of Fujian Medical University, Fuzhou 350001, China;  2.Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China
  • Received:2017-07-24 Revised:2017-10-10 Online:2017-04-06 Published:2018-04-06
  • Contact: QUAN Song E-mail:quansong@smu.edu.cn

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摘要：

目的 探讨卵丘颗粒细胞骨形态发生蛋白（BMP）-15、BMP-6信号通路分子表达对受精卵发育能力的影响。 方法 收集2012年4月~2014年6月，因男性不育症行卵泡浆内单精子显微注射技术（ICSI）助孕的119例病例，456份单个卵子的卵丘颗粒细胞，采用免疫细胞化学，Western blotting方法检测颗粒细胞BMP-15、BMP-6信号通路分子蛋白的表达，采用Real-time PCR方法检测BMP-15、BMP-6信号分子基因表达。比较分析卵子ICSI后发育形成囊胚组与非囊胚组卵丘颗粒细胞BMP-15、BMP-6信号分子表达差异。 结果 对两组卵丘颗粒细胞，进行了免疫细胞化学，Real-time PCR，Western blotting分析，结果发现，BMP-15、BMP-6、AKL2、AKL6、BMPRⅡ、Smad1、Samd4、Smad5、Samd8及E盒结合锌指蛋白1（ZEB1）在两组颗粒细胞中均有表达，其中BMP-15、BMP-6、AKL2、AKL6、BMPRⅡ、Smad1 Samd4及ZEB1表达，组间比较差异具有显著性（经Bonferroni校正P＜0.005）。而Smad5、Samd8 表达，组间比较差异无显著性（P＞0.05）。 结论 卵丘颗粒细胞BMP-15、BMP-6、AKL2、AKL6、BMPRⅡ、Smad1、Samd4及ZEB1信号通路分子可能通过信号通路的调控作用，对受精卵发育能力产生影响。

关键词: 囊胚, 卵丘颗粒细胞, 骨形态发生蛋白, 卵泡浆内单精子显微注射技术, 女性

Abstract:

Objective The purpose of this study is to evaluate the effect of oophorus cumulus granulosa cells bone morphogenetic protein(BMP)-15，BMP-6 signaling molecules expression on blastocyst information after intractoplasmic sperm injection(ICSI), and assess the associations with the developmental ability of oocytes through ICSI. Methods A total of 119 women receiving ICSI due to male infertility in Fujian Maternity and Children Health Hospital during the period from April 2012 to June 2014 were enrolled in the study.456 oophorus cumulus granulosa cell samples were collected from each oocyte for ICSI. BMP-15, BMP-6, AKL2, AKL6, BMPRⅡ, Smad1, Samd4, Smad5, Samd8 and zinc finger E-box-binding protein(ZEB1) signaling molecules were determined by immunocytochemistry staining and Western blotting assay, while the mRNA expression was quantified using Real-time PCR assay. The molecules expression differences between blastocyst and non-blastocyst groups after ICSI were compared. Results The intensities of immunohistochemical positive staining for BMP15, BMP-6,AKL2,AKL6, BMPRⅡ, Smad1,2,5,and 8 and ZEB1 were at various levels and the positive staining were distributed in oophorus cumulus cells. The intensities of immunohistochemical staining for BMP-15,BMP-6, AKL2,6, BMPRⅡ,Smad1,4 and ZEB1 were higher in the blstocyst group than those in non-blastocyst group from ICSI (P＞0.05). While there no significant differences for staining intensities of Smad5, Smad8 between two groups (P＞0.05). While there was no significant differences of Smad5, Samd8 expression were found between two groups (P＞0.05). Conclusion BMP-15, BMP-6, AKL2, AKL6, BMPRⅡ, Smad1, Samd4 and ZEB1 signaling molecules may play an important role in the development of oocytes and promotion of blastocyst information from embryo culture after oocyte fertilization through ICSI.

Key words: Blastocyst, Cumulus granulosa cell, Bone morphogenetic protein, Intractoplasmic sperm injection, Female