Sabés-Alsina Maria, Planell Núria, Gil Sílvia, Tallo-Parra Oriol, Maya-Soriano Maria José, Taberner Ester, Piles Miriam, Sabés Manel, Lopez-Bejar Manel
Department of Animal Health and Anatomy,Universitat Autònoma de Barcelona,08193 Bellaterra,Spain.
Department of Gastroenterology and Bioinformatics Platform,CIBERehd,Barcelona,Spain.
Zygote. 2016 Oct;24(5):707-13. doi: 10.1017/S0967199415000726. Epub 2016 Mar 11.
The resazurin reduction test (RRT) is a useful technique to assess the metabolic rate of sperm cells. RRT depends on the ability of metabolically active cells to reduce the non-fluorescent dye resazurin to the fluorescent resorufin. The aim of this study was to develop a vital fluorometric method to evaluate metabolic activity of rabbit sperm cells. Twenty-five rabbit males were included in the study. Viability and morphology, motility and metabolic activity were evaluated using an eosin-nigrosin staining, a computer-assisted semen analysis (CASA) and the RRT, respectively. Spearman rank correlation analysis was used to determine the correlation between RRT and semen parameters. After evaluation, a concentration of 10 × 106 sperm cells/ml was selected for further experiments with RRT. No significant correlation was found between the RRT results and the motility parameters. However, after RRT a significant positive correlation between relative fluorescence units and the percentage of alive spermatozoa (r = 0.62; P = 0.001) and a negative one with the percentage of sperm cells with acrosomic abnormalities (r = -0.45; P < 0.05) were detected. The vital assessment of metabolic rate of sperm cells by RRT could provide more information about semen quality than other routine semen analysis, correlating with sperm viability and acrosome status information.
刃天青还原试验(RRT)是一种评估精子细胞代谢率的有用技术。RRT取决于代谢活跃的细胞将非荧光染料刃天青还原为荧光试卤灵的能力。本研究的目的是开发一种重要的荧光测定方法来评估兔精子细胞的代谢活性。25只雄兔纳入本研究。分别使用伊红-黑色素染色、计算机辅助精液分析(CASA)和RRT评估活力与形态、运动性和代谢活性。采用Spearman等级相关分析确定RRT与精液参数之间的相关性。评估后,选择10×10⁶个精子细胞/ml的浓度用于RRT的进一步实验。RRT结果与运动参数之间未发现显著相关性。然而,RRT后检测到相对荧光单位与活精子百分比之间存在显著正相关(r = 0.62;P = 0.001),与顶体异常精子细胞百分比之间存在负相关(r = -0.45;P < 0.05)。与其他常规精液分析相比,通过RRT对精子细胞代谢率进行重要评估可以提供更多关于精液质量的信息,与精子活力和顶体状态信息相关。