Ciepiela Przemysław, Bączkowski Tomasz, Drozd Arleta, Kazienko Anna, Stachowska Ewa, Kurzawa Rafał
Department of Reproductive Medicine and Gynecology, Pomeranian Medical University, Szczecin-Police, Zachodniopomorskie, Poland.
Department of Reproductive Medicine and Gynecology, Pomeranian Medical University, Szczecin-Police, Zachodniopomorskie, Poland; VitroLive Fertility Clinic, Szczecin, Zachodniopomorskie, Poland.
PLoS One. 2015 Mar 12;10(3):e0119087. doi: 10.1371/journal.pone.0119087. eCollection 2015.
To evaluate human oocyte ability to undergo fertilization and subsequent preimplantation embryonic development in relation to a wide panel of follicular fluid (FF) arachidonic acid derivatives (AAD) and linoleic acid derivatives (LAD) of prospectively selected patients undergoing intracytoplasmic sperm injection (ICSI).
Study was designed as a two center (a university clinic and a private clinic) prospective study. 54 women of 181 consecutive couples undergoing ICSI were prospectively found to be eligible for analysis. 'One follicle - one retrieved oocyte - one resulting embryo' approach was used. Each individual follicle was aspirated independently and matched to an oocyte growing in this particular follicular milieu. FF samples were assessed for AAD and LAD by high-performance liquid chromatography; additionally, activity of secretory phospholipase A (sPLA2) was determined by enzyme-linked immunosorbent assay.
Increased activity of sPLA2 and significantly higher AAD and LAD levels were found in FF of oocytes that did not show two pronuclei or underwent degeneration after ICSI in comparison to oocytes with the appearance of two pronuclei. Receiver operating characteristics curve analysis identified acids with the highest sensitivity and specificity: 5oxo-hydroxyeicosatetraenoic, 16-hydroxyeicosatetraenoic, 9-hydroxyoctadecadieneoic and 12-hydroxyeicosatetraenoic. No significant differences between AAD and LAD related to embryo quality were found.
CONCLUSIONS/SIGNIFICANCE: Our study demonstrates for the first time that elevated concentrations of AAD and LAD in FF at the time of oocyte retrieval significantly decrease the ability of oocytes to form pronuclei after ICSI. This may serve as a new tool for non-invasive assessment of oocyte developmental capacity. However, levels of AAD and LAD are not associated with subsequent embryo quality or pregnancy rate, and therefore more studies are needed to determine their usefulness in human IVF procedure.
评估在接受卵胞浆内单精子注射（ICSI）的前瞻性选择患者中，人类卵母细胞进行受精及随后的植入前胚胎发育的能力与一系列卵泡液（FF）花生四烯酸衍生物（AAD）和亚油酸衍生物（LAD）之间的关系。
本研究设计为一项两中心（一所大学诊所和一所私立诊所）的前瞻性研究。在181对连续接受ICSI的夫妇中，前瞻性地发现54名女性符合分析条件。采用“一个卵泡 - 一个获取的卵母细胞 - 一个产生的胚胎”方法。每个单独的卵泡被独立抽吸，并与在该特定卵泡环境中生长的卵母细胞匹配。通过高效液相色谱法评估FF样本中的AAD和LAD；此外，通过酶联免疫吸附测定法测定分泌型磷脂酶A（sPLA2）的活性。
与出现两个原核的卵母细胞相比，在ICSI后未显示两个原核或发生退化的卵母细胞的FF中，发现sPLA2活性增加以及AAD和LAD水平显著更高。受试者工作特征曲线分析确定了具有最高敏感性和特异性的酸：5-氧代-羟基二十碳四烯酸、16-羟基二十碳四烯酸、9-羟基十八碳二烯酸和12-羟基二十碳四烯酸。未发现与胚胎质量相关的AAD和LAD之间存在显著差异。
结论/意义：我们的研究首次表明，在卵母细胞采集时FF中AAD和LAD浓度升高会显著降低ICSI后卵母细胞形成原核的能力。这可能作为一种非侵入性评估卵母细胞发育能力的新工具。然而，AAD和LAD水平与随后的胚胎质量或妊娠率无关，因此需要更多研究来确定它们在人类体外受精程序中的有用性。
