Ghanbari Hamideh, Keshtgar Sara, Kazeroni Marjaneh
Department of Physiology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Shiraz Infertility Center, Shiraz, Iran.
Iran J Med Sci. 2018 Jan;43(1):18-25.
Low levels of reactive oxygen species (ROS) and calcium are necessary for sperm function. NADPH oxidase 5 (NOX5) is a membrane enzyme which produces ROS. This enzyme is dependent on calcium for its activity. We investigated the importance of NOX5 and an important calcium channel (CatSper) on sperm function.
This laboratory study was done in Shiraz, Iran, 2016. Normal semen samples (n=24) were washed and diluted to 20×10 sperm/mL. The diluted samples were divided into 8 groups, containing Ham's F-10 (control group), 2 µM of NNC (CatSper channel inhibitor), 1 µM DPI (NOX5 inhibitor), and NNC+DPI. The other 4 groups were the same as the 1st ones, except that they contained 1 µM of progesterone. Motility assessment was done by VT-Sperm 3.1. Acrosome status was monitored with acrosome-specific FITC-PSA using fluorescent microscopy. Sperm viability was assessed by Eosin Y. Statistical analysis was performed using SPSS 16 software. The comparison between the groups was done using the one-way ANOVA, followed by Tukey. A P<0.05 was considered significant.
The percentage of motile sperm, sperm velocity, and viability decreased significantly in the groups containing NNC. DPI reduced sperm progressive motility only in the progesterone-stimulated condition. Progesterone induced acrosome reaction, but this effect was inhibited by NNC and DPI.
CatSper had a prominent role in the motility, acrosome reaction, and viability of the human sperm. The function of NOX5 was important only in the stimulated sperm. We conclude that CatSper has a more prominent role than NOX5 activity. The functional relation between NOX5 and CatSper is not clear but is very probable.
低水平的活性氧（ROS）和钙对于精子功能是必需的。NADPH氧化酶5（NOX5）是一种产生ROS的膜酶。该酶的活性依赖于钙。我们研究了NOX5和一种重要的钙通道（CatSper）对精子功能的重要性。
这项实验室研究于2016年在伊朗设拉子进行。将正常精液样本（n = 24）洗涤并稀释至20×10⁶精子/mL。将稀释后的样本分为8组，包括哈姆氏F - 10（对照组）、2 μM的NNC（CatSper通道抑制剂）、1 μM DPI（NOX5抑制剂）以及NNC + DPI。另外4组与第一组相同，只是含有1 μM的孕酮。使用VT - Sperm 3.1进行活力评估。使用荧光显微镜通过顶体特异性FITC - PSA监测顶体状态。用伊红Y评估精子活力。使用SPSS 16软件进行统计分析。组间比较采用单因素方差分析，随后进行Tukey检验。P < 0.05被认为具有显著性。
含有NNC的组中，活动精子的百分比、精子速度和活力显著降低。DPI仅在孕酮刺激的条件下降低精子的前向运动能力。孕酮诱导顶体反应，但这种作用被NNC和DPI抑制。
CatSper在人类精子的活力、顶体反应和活力方面发挥着重要作用。NOX5的功能仅在受刺激的精子中重要。我们得出结论，CatSper比NOX5活性发挥着更重要的作用。NOX5和CatSper之间的功能关系尚不清楚，但很可能存在。
