纤连蛋白作为辅助生殖技术中人类精子筛选的一种新生物标志物。

Fibronectin as a new biomarker for human sperm selection in assisted reproductive technology.

作者信息

Heidari Mahnaz, Darbandi Sara, Darbandi Mahsa, Akhondi Mohammad Mehdi, Sadeghi Mohammad Reza

机构信息

Department of Embryology and Andrology, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

出版信息

Turk J Urol. 2019 Jan 22;45(2):83-90. doi: 10.5152/tud.2019.30660. Print 2019 Mar.

DOI:10.5152/tud.2019.30660

PMID:30875286

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6368043/

Abstract

OBJECTIVE

Fibronectin (FN) is a multifunctional diametric glycoprotein on the surface of the sperm that plays an important role in the sperm-oocyte interaction and fertilization process. The aim of the present study was to assess the FN levels as a sperm surface biomarker for sperm selection in assisted reproductive technology.

MATERIAL AND METHODS

Polyclonal antibody against human FN was produced in rabbit. Its quality, purity, and immune reactivity were assessed by SDS-PAGE and western blot. In addition, the presence of FN on the sperm surface was assessed through immunocytochemistry and flow cytometry. The amount of FN and the sperm quality were assessed in normozoospermia (N) (42 men) and asthenoteratozoospermia (AT) (72 men) groups through sperm chromatin dispersion (SCD), sperm chromatin structure assay (SCSA), and chromatin maturation index (CMI).

RESULTS

The results showed the distribution of FN protein on the equatorial region of the human sperm surface. In addition, the FN levels were found to have a significant difference between the two groups with 24.64±9.08% in N and 16.90±7.27% in AT (p≤0.0001). In addition, FN level negatively correlated with SCD (p≤0.0001), SCSA (p≤0.0001), and CMI (p≤0.001). Threshold values of FN level and DNA fragmentation index (DFI) percentage were 16 and 30 and were identified as cut-off values to determine the N group with a specificity of 83.3% and 81.0% and a sensitivity of 16.8% and 19.0%, respectively. The specificity and sensitivity of FN-DFI were 91.2% and 8.8%, respectively.

CONCLUSION

It appears that FN can be used for the selection of sperm with suitable quality, although future studies are recommended.

摘要

目的

纤连蛋白（FN）是精子表面的一种多功能二聚体糖蛋白，在精子与卵母细胞相互作用及受精过程中发挥重要作用。本研究旨在评估纤连蛋白水平作为辅助生殖技术中精子筛选的一种精子表面生物标志物。

材料与方法

用兔制备抗人纤连蛋白的多克隆抗体。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）和蛋白质免疫印迹法评估其质量、纯度和免疫反应性。此外，通过免疫细胞化学和流式细胞术评估精子表面纤连蛋白的存在情况。通过精子染色质扩散试验（SCD）、精子染色质结构分析（SCSA）和染色质成熟指数（CMI），对正常精子症（N）组（42名男性）和弱畸精子症（AT）组（72名男性）的纤连蛋白含量和精子质量进行评估。

结果

结果显示纤连蛋白在人精子表面赤道区域分布。此外，发现两组之间纤连蛋白水平存在显著差异，N组为24.64±9.08%，AT组为16.90±7.27%（p≤0.0001）。此外，纤连蛋白水平与SCD（p≤0.0001）、SCSA（p≤0.0001）和CMI（p≤0.001）呈负相关。纤连蛋白水平和DNA碎片化指数（DFI）百分比的阈值分别为16和30，被确定为区分N组的临界值，特异性分别为83.3%和81.0%，敏感性分别为16.8%和19.0%。纤连蛋白 - DFI的特异性和敏感性分别为91.2%和8.8%。