Central Laboratory, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai, Shandong, China.
Reproductive Center, Tianjin Aiwei Hospital, Tianjin, China.
Andrology. 2020 Mar;8(2):407-420. doi: 10.1111/andr.12687. Epub 2019 Jul 31.
Seminal plasma is a promising diagnostic fluid for male infertility. In assisted reproduction, the seminal plasma-based characteristics of normozoospermic men achieving successful clinical pregnancy through rescue intracytoplasmic sperm injection after in vitro fertilization failure remain unclear.
To identify potential seminal plasma proteins to contribute to a new understanding of unexplained male factor infertility.
An approach with isobaric tags for relative and absolute quantification labeling coupled with liquid chromatography matrix-assisted laser desorption ionization mass spectrometry was applied to investigate differentially expressed proteins in the seminal plasma of a rescue intracytoplasmic sperm injection pregnancy group versus an in vitro fertilization pregnancy group of normozoospermic men.
RESULT(S): The present work revealed seventy-three differentially expressed seminal plasma proteins between the in vitro fertilization and rescue intracytoplasmic sperm injection groups. Forty-five proteins were upregulated, and 28 proteins were downregulated in the rescue intracytoplasmic sperm injection group compared with the in vitro fertilization group. Bioinformatics analyses showed that these altered proteins were involved in various functions, including the kallikrein-related proteolytic cascade, immune response, and heparin binding. Furthermore, the validity of the proteomic results was verified by Western blot analysis of the proteins (lactoferrin [LTF], fibronectin [FN1], creatine kinase B type [CKB], kallikrein-2 [KLK2], aminopeptidase N [ANPEP], extracellular matrix protein 1 [ECM1], glycodelin [PAEP], alpha-1-antitrypsin [SERPINA1], and semenogelin-1 [SEMG1]) and immunofluorescence. Moreover, 16% of the seminal plasma proteins identified in the present work have not been reported in previous studies.
This panel of altered seminal plasma proteins associated with unexplained male factor infertility might have clinical relevance and may be useful in the diagnosis and prognosis of idiopathic infertility in in vitro fertilization.
Our work not only provides a new complementary high-confidence dataset of seminal plasma proteins but also shines new light onto the molecular characteristics of seminal plasma from normozoospermic men with different assisted reproductive outcomes.
精液是男性不育症有前途的诊断液。在辅助生殖中，对于体外受精失败后通过卵胞浆内精子注射挽救治疗获得成功临床妊娠的正常精子男性，其基于精液的特征仍然不清楚。
确定潜在的精液蛋白，以帮助我们更好地理解不明原因的男性因素不育症。
采用相对和绝对定量标记的同位素标记与液相色谱-基质辅助激光解吸电离飞行时间质谱联用的方法，研究正常精子男性体外受精和卵胞浆内精子注射挽救治疗妊娠组的精液中差异表达蛋白。
本研究揭示了体外受精和卵胞浆内精子注射挽救治疗组之间 73 种差异表达的精液蛋白。与体外受精组相比，卵胞浆内精子注射组中 45 种蛋白上调，28 种蛋白下调。生物信息学分析表明，这些改变的蛋白参与了各种功能，包括激肽释放酶相关蛋白水解级联、免疫反应和肝素结合。此外，通过蛋白质（乳铁蛋白[LTF]、纤维连接蛋白[FN1]、肌酸激酶 B 型[CKB]、激肽释放酶-2[KLK2]、氨肽酶 N[ANPEP]、细胞外基质蛋白 1[ECM1]、糖蛋白 1[PAEP]、α-1-抗胰蛋白酶[SERPINA1]和 semenogelin-1[SEMG1]）的 Western blot 分析和免疫荧光验证了蛋白质组学结果的有效性。此外，本研究中鉴定的 16%的精液蛋白以前的研究中没有报道过。
与不明原因的男性因素不育相关的这种改变的精液蛋白可能具有临床相关性，并且可能有助于体外受精中特发性不育症的诊断和预后。
我们的工作不仅提供了一个新的补充性的、高可信度的精液蛋白数据集，而且为具有不同辅助生殖结局的正常精子男性精液的分子特征提供了新的认识。
