Kaeoket K, Persson E, Dalin A M
Department of Obstetrics and Gynaecology, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Uppsala, Sweden.
J Vet Med A Physiol Pathol Clin Med. 2003 May;50(4):169-78. doi: 10.1046/j.1439-0442.2003.00532.x.
This study investigates the distribution of leucocytes, CD2+, CD4+, CD8+ lymphocyte subpopulations and MHC class II expressing cells in the sow endometrium following post-ovulatory insemination in relation to clinical findings and pregnancy outcome. Crossbred multiparous sows were inseminated once either at 15-20 h after ovulation [experiment 1, slaughtered at 20-25 h (5-6 h after artificial insemination (AI), group 1-A, n = 4), at 70 h after ovulation (group 1-B, n = 4), on day 11 (group 1-C, n = 4, first day of standing oestrus = day 1) or on day 19 (group 1-D, n = 4)] or 30 h after ovulation [experiment 2, slaughtered at 5-6 h after AI (group 2-A, n = 4) or on day 19 (group 2-D, n = 3)]. The uterine horns were flushed to control for the presence of spermatozoa and neutrophils and/or for recovery of oocytes and/or embryos. Mesometrial uterine samples were plastic embedded and stained. Cryofixed uterine samples were analysed by immunohistochemistry using mAbs to lymphocyte subpopulations and MHC class II molecules. Light microscopy was used to examine surface (SE) and glandular epithelia (GE), and connective tissue layers, both subepithelially (SL) and glandular (GL). In experiment 1, group 1-A, only one sow had spermatozoa in the utero-tubal junction (UTJ). Marked/moderated numbers of neutrophils and spermatozoa were observed in the flushings of two sows. In group 1-B, altogether 23 of 48 oocytes were cleaved. Day 11 (1-C), embryos with small diameter were observed. Day 19 (1-D), no embryos were found but small pieces of foetal membrane were observed in one of the sows. In group 1-A, large numbers of neutrophils were found within the SE and SL but with high individual variation. For T lymphocyte subpopulations, in the SE, most CD2+ cells were found in group 1-A. For both SE and GE in all groups, the number of CD8+ cells was significantly larger than that of CD4+ cells. In experiment 2, group 2-A, no sow had spermatozoa in the UTJ or in the uterine flushings. At day 19, no sow was pregnant. In group 2-A, large numbers of neutrophils were found within the SE and SL but with high individual variation. At day 19, high E2 levels showed a hormonal prooestrous stage but the endometrial neutrophil infiltration normally expected at pro-oestrus was absent. In conclusion, post-ovulatory insemination (about 18 h after ovulation) resulted in impaired spermatozoa transport within the uterus and embryonic degeneration. In sows post-ovulatory inseminated at a later stage (30 h after ovulation), no sow was pregnant. In both experiments, disturbed immune cell patterns were observed in some individuals.
本研究调查了排卵后授精的母猪子宫内膜中白细胞、CD2⁺、CD4⁺、CD8⁺淋巴细胞亚群以及表达MHC II类分子的细胞的分布情况，并将其与临床发现和妊娠结局相关联。杂种经产母猪在排卵后15 - 20小时（实验1，在20 - 25小时[人工授精（AI）后5 - 6小时，1 - A组，n = 4]、排卵后70小时（1 - B组，n = 4）、第11天（1 - C组，n = 4，静立发情第一天 = 第1天）或第19天（1 - D组，n = 4）进行一次授精）或排卵后30小时（实验2，在AI后5 - 6小时（2 - A组，n = 4）或第19天（2 - D组，n = 3）进行一次授精）进行授精。冲洗子宫角以检查是否存在精子和中性粒细胞，以及/或者回收卵母细胞和/或胚胎。将子宫系膜侧子宫样本进行塑料包埋和染色。使用针对淋巴细胞亚群和MHC II类分子的单克隆抗体，通过免疫组织化学对冷冻固定的子宫样本进行分析。用光学显微镜检查表面上皮（SE）和腺上皮（GE），以及上皮下（SL）和腺内（GL）的结缔组织层。在实验1的1 - A组中，仅1头母猪在子宫输卵管连接处（UTJ）有精子。在2头母猪的冲洗液中观察到大量/中等数量的中性粒细胞和精子。在1 - B组中，48个卵母细胞中共有23个发生了卵裂。在第11天（1 - C组），观察到直径较小的胚胎。在第19天（1 - D组），未发现胚胎，但在1头母猪中观察到小块胎膜。在实验1的1 - A组中，在SE和SL内发现大量中性粒细胞，但个体差异较大。对于T淋巴细胞亚群，在SE中，1 - A组中发现的CD2⁺细胞最多。在所有组的SE和GE中，CD8⁺细胞的数量均显著多于CD4⁺细胞。在实验2的2 - A组中，没有母猪在UTJ或子宫冲洗液中有精子。在第19天，没有母猪怀孕。在2 - A组中，在SE和SL内发现大量中性粒细胞，但个体差异较大。在第19天，高雌激素水平表明处于激素性发情前期，但发情前期通常预期的子宫内膜中性粒细胞浸润不存在。总之，排卵后授精（排卵后约18小时）导致子宫内精子运输受损和胚胎退化。在排卵后期（排卵后30小时）进行排卵后授精的母猪中，没有母猪怀孕。在两个实验中，在一些个体中观察到免疫细胞模式紊乱。
