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 全松* 康跃凡 余爱丽 林元

doi:10.16098/j.issn.0529-1356.2018.02.016

PDF(814 KB)

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

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

作者信息 +

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

Author information +

文章历史 +

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。

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.

导出引用

林典梁全松康跃凡余爱丽林元. 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[J]. 解剖学报. 2018, 49(2): 233-239 https://doi.org/10.16098/j.issn.0529-1356.2018.02.016

LIN Dian-liang QUAN Song KANG Yue-fan YU Ai-li LIN Yuan.

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

[J]. Acta Anatomica Sinica. 2018, 49(2): 233-239 https://doi.org/10.16098/j.issn.0529-1356.2018.02.016

参考文献

［1］Sadr SZ, Ebrahimi B, Shahhoseini M, et al. Mouse preantral follicle development in two-dimensional and three-dimensional culture systems after ovarian tissue vitrification ［J］. Eur J Obstet Gynecol Reprod Biol, 2015,194:206-211.

［2] Sugiura K, Su YQ, Eppig JJ. Does bone morphogenetic protein 6 (BMP-6) affect female fertility in the mouse［J］? Biol Reprod, 2012, 83(6):997-1004.

［3］Lin D, Ran J, Chen C, et al. Frozen two day 3 embryos and subsequently produced blastocysts by vitrification: advantages for IVF/ICSI patients at high risk of ovarian hyperstimulation syndrome［J］. Int J Clin Exp Med, 2016, 9(3):6986-6999.

［4］Sirard MA,Blondin P. Oocyte maturation and IVF in cattle［J］．Anim Reprod Sci, 1996, 42(14)：417-426．

［5］Wang ZhW, Yue ShL, Zhu JW, et al. Progeny of 2-cell embryo blastomeres distribute in mouse blastocyst randomly［J］. Acta Anatomica Sinica, 2010,41(1):124-127．(in Chinese)

王中伟,岳顺利,朱佳伟,等.小鼠2-细胞胚胎卵裂球后代在囊胚中随机分布［J］. 解剖学报, 2010,41(1):124-127.

［6］Lin D, Ran J, Zhu S, et al. Effect of GOLPH3 on cumulus granulosa cell apoptosis and ICSI pregnancy outcomes［J］. Sci Rep, 2017,7(1):7863.

［7］Soslow RA, Dannenberg AJ, Rush D, et al. Cox-2 is expressed in human pulmonary, colonic, and mammary tumors［J］. Cancer, 2000, 89(12):2637-2645.

［8］Wu YT,Tang L，Cai J，et al．High bone morphogenetic protein-1 5 level in follicular fluid is associated with high quality oocyte and subsequent embryonic development［J］．Hum Reprod，2007，22(6)：1526-1531.

［9］Shi J，Yoshino O，Osuga Y, et al．Bone morphogenetic protein-6 stimulates gene expression of follicle-stimulatinghormone receptor,inhibin/activin beta subunits，and anti-Mullerian hormone in human granulosa cells［J］．Fertil Steril，2009，92(5)：1794-1798.

［10］Gónzalez A, RamírezLorca R, Calatayud C, et al. Association of genetic markers within the BMP-15 Gene with an ovulation and infertility in women with polycystic ovary syndrome ［J］. Fertil Steril, 2008, 90(2): 447-449.

［11］Rossetti R, Di Pasquale E, Marozzi A, et al. BMP15 mutations associated with primary ovarian insufficiency cause a defective production of bioactive protein ［J］. Hum Mutat, 2009, 30(5):804-810.

［12］Wu YT, Tang L, Cai J, et al. High bone morphogenetic protein-15 level in follicular fluid is associated with high quality oocyte and subsequent embryonic development ［J］. Hum Reprod, 2007, 22(6):1526-1531.

［13］Guiyu Z，Bingran G， Dengke P，et al．Expression of bone morphogentic proteins and receptors in porcine cumulus-oocyte complexes during in vitro maturation［J］．Ani Reprod Sci，2008, 104(2-4): 275-283.

［14］Otsuka F. Multifunctional bone morphogenetic protein system in endocrinology［J］. Acta Med Okayama, 2013, 67(2):75-86.

［15］Shi J, Yoshino O，Osuga Y, et，a1．Growth differentiation factor 3 is induced by bone morphogenetic protein 6(BMP-6)and BMP-7 and increases luteinizing hormone receptor meSsenger RNA expression in human cells［J］．Fertil Steril, 2012，97(4):979-983.

［16］Da silva faria T, De bittencourt BRASIL, Fsampaio FJ, et al．Effects of maternal under-nutrition during lactation on estrogen and androgen receptor expressions in rat ovary at puberty［J］．Nutrition，2010, 26(10)：993-999．

［17］Emori C, Sugiura K. Role of oocyte-derived paracrine factors in follicular development ［J］. Anim Sci J, 2014,85(6):627-633.

［18］Zhang Y, Zhang DY, Zhao YF, et al．Analysis and characterization of the functional TGF-βreceptors required for BMP-6 induced osteogenic differentiation of mesenchymal progenitor cells［J］．BMB Rep，2013，46(2)：107-112.

［19］Miyazawa K, Shinozaki M, Hara T, et al．Two major Smad pathways in TGF-beta superfamily signalling［J］．Genes Cells，2002，7(12)：1191-1204.

［20］Moore RK, Otsuka F, Shimasaki S．Molecular basis of bone morphogenetic protein-1 5 signaling in granulosa cells［J］．J Biol Chem，2003，278(1)：304-310.

［21］Jin T, George Fantus I, Sun J. Wnt and beyond Wnt: multiple mechanism control the transcriptional property of beta-caternin［J］. Cell Singnal, 2008，20(10): 1697-1704.

［22］Li HY, Pei HT, Ru LJ. Effect of bone morphogenetic protein-7 on expression of Caspase-3 in rats with cerebral ischemic reperfusion injury［J］. Acta Anatomica Sinica, 2012, 43(3): 335-339．(in Chinese)

李红云, 裴海涛, 汝丽娟.骨形态发生蛋白-7对大鼠脑缺血再灌注后Caspase-3表达的影响［J］.解剖学报, 2012, 43(3): 335-339．