Khatun Amena, Kang Kyu-Ho, Ryu Do-Yeal, Rahman Md Saidur, Kwon Woo-Sung, Pang Myung-Geol
Department of Animal Science and Technology, Chung-Ang University, Anseong, Gyeonggi-do 456-756, South Korea; Department of Agriculture, Faculty of Science, Noakhali Science and Technology University, Noakhali, 3814, Bangladesh.
Department of Animal Science and Technology, Chung-Ang University, Anseong, Gyeonggi-do 456-756, South Korea.
Theriogenology. 2018 Sep 15;118:182-189. doi: 10.1016/j.theriogenology.2018.06.005. Epub 2018 Jun 12.
Aminopeptidase N (APN) is defined as a multifunctional enzyme, which regulate cellular physiology of a wide variety of cells in human. Earlier studies reported that mammalian semen shares this common enzyme as a major protein of seminal plasma that has correlation with male fertility, while the regulatory mechanisms of APN in spermatozoa are still far from being well understood. Present study was designed to investigate the role of APN in biological and chemical functions of spermatozoa using an in vitro antagonistic approach. Results showed that lower APN activity in sperm culture medium significantly increased sperm motility and the percentage of high speed spermatozoa and decreased the percentage of slow speed spermatozoa after a dose dependent inhibitor treatment (10, 100, and 1000 μM leuhistin) on epididymal mouse spermatozoa in a capacitating media for 90 min. Both 100 μM and 1000 μM decreased APN activity, while only 1000 μM decreased cell viability and increased PKA activity significantly compared to control. Nonetheless capacitation status, acrosome reaction status, and lactate dehydrogenase activity were not affected. Intriguingly, the treatment affected embryonic development through decreasing tyrosine phosphorylation of proteins and increasing reactive oxygen species levels. Further in silico analysis revealed associated regulatory proteins, which have critical functional role for male fertility.
氨肽酶N（APN）被定义为一种多功能酶，它调节人体多种细胞的细胞生理功能。早期研究报道，哺乳动物精液中存在这种常见酶，它是精浆的主要蛋白质，与男性生育能力相关，但APN在精子中的调节机制仍远未被完全理解。本研究旨在采用体外拮抗方法研究APN在精子生物学和化学功能中的作用。结果表明，在体外获能培养基中对附睾小鼠精子进行剂量依赖性抑制剂（10、100和1000μM亮抑酶肽）处理90分钟后，精子培养基中较低的APN活性显著提高了精子活力和高速精子的百分比，并降低了低速精子的百分比。100μM和1000μM均降低了APN活性，但与对照组相比，只有1000μM显著降低了细胞活力并提高了蛋白激酶A（PKA）活性。尽管如此，获能状态、顶体反应状态和乳酸脱氢酶活性均未受到影响。有趣的是，该处理通过降低蛋白质的酪氨酸磷酸化水平和提高活性氧水平影响胚胎发育。进一步的计算机分析揭示了相关的调节蛋白，它们对男性生育能力具有关键的功能作用。
