Marey Mohamed A, Liu Jinghui, Kowsar Rasoul, Haneda Shingo, Matsui Motozumi, Sasaki Motoki, Takashi Shimizu, Hayakawa Hiroyuki, Wijayagunawardane Missaka P B, Hussein Fekry M, Miyamoto Akio
Graduate School of Animal and Food Hygiene, Obihiro University of Agriculture and Veterinary Medicine, Obihiro 080-8555, Japan.
Reproduction. 2013 Dec 24;147(2):211-9. doi: 10.1530/REP-13-0375. Print 2014 Feb.
This study aimed to investigate the presence of polymorphonuclear neutrophils (PMNs) in bovine oviduct fluid under physiological conditions and to determine the possible role of bovine oviduct epithelial cells (BOECs) in the regulation of the phagocytic activity of PMNs for sperm. During the pre-ovulatory stage, PMNs were identified in the bovine oviduct fluid in relatively constant numbers. In our experiments, PMNs were incubated for 4 h with the supernatant of cultured BOECs stimulated for 24 h by LH (10 ng/ml). Phagocytosis was then assayed by co-incubation of these PMNs with sperm treated to induce capacitation. The BOEC supernatant significantly suppressed sperm phagocytosis by PMNs, and the LH-stimulated BOEC supernatant further suppressed phagocytosis. Importantly, in the BOEC culture, LH stimulated the secretion of prostaglandin E2 (PGE2), which dose-dependently (10(-6), 10(-7), and 10(-8) M) suppressed sperm phagocytosis by PMNs. Furthermore, a PGEP2 receptor antagonist significantly abrogated the inhibition of phagocytosis by the LH-stimulated BOEC supernatant. Additionally, using scanning electron microscopy, incubation of PMNs with either PGE2 or LH-stimulated BOEC supernatant before phagocytosis was found to prevent the formation of DNA-based neutrophil extracellular traps for sperm entanglement. The results indicate that sperm are exposed to PMNs in the oviduct and PGE2 released into the oviduct fluid after LH stimulation may play a major role in the suppression of the phagocytic activity of PMNs for sperm via interaction with EP2 receptors. Thus, the bovine oviduct provides a PGE2-rich microenvironment to protect sperm from phagocytosis by PMNs, thereby supporting sperm survival in the oviduct. Free Japanese abstract A Japanese translation of this abstract is freely available at http://www.reproduction-online.org/content/147/2/211/suppl/DC1.
本研究旨在调查生理条件下牛输卵管液中多形核中性粒细胞(PMNs)的存在情况,并确定牛输卵管上皮细胞(BOECs)在调节PMNs对精子吞噬活性中的可能作用。在排卵前期,牛输卵管液中可检测到数量相对恒定的PMNs。在我们的实验中,将PMNs与经促黄体生成素(LH,10 ng/ml)刺激24小时的培养BOECs的上清液孵育4小时。然后通过将这些PMNs与经诱导获能处理的精子共同孵育来检测吞噬作用。BOEC上清液显著抑制了PMNs对精子的吞噬作用,而经LH刺激的BOEC上清液进一步抑制了吞噬作用。重要的是,在BOEC培养中,LH刺激了前列腺素E2(PGE2)的分泌,其以剂量依赖性方式(10⁻⁶、10⁻⁷和10⁻⁸ M)抑制了PMNs对精子的吞噬作用。此外,一种PGE2受体拮抗剂显著消除了经LH刺激的BOEC上清液对吞噬作用的抑制。另外,使用扫描电子显微镜观察发现,在吞噬作用前将PMNs与PGE2或经LH刺激的BOEC上清液孵育可防止形成用于精子缠绕的基于DNA的中性粒细胞胞外陷阱。结果表明,精子在输卵管中会接触到PMNs,LH刺激后释放到输卵管液中的PGE2可能通过与EP2受体相互作用在抑制PMNs对精子的吞噬活性中起主要作用。因此,牛输卵管提供了一个富含PGE2的微环境来保护精子免受PMNs的吞噬,从而支持精子在输卵管中的存活。免费日文摘要 本摘要的日文翻译可在http://www.reproduction-online.org/content/147/2/211/suppl/DC1免费获取。