Reilas Tiina, Rivera Del Alamo Maria Montserrat, Liepina Evija, Yeste Marc, Katila Terttu
Green Technology, Natural Resources Institute Finland, Ypäjä, Finland.
Animal Medicine and Surgery Department, Unit of Reproduction, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Spain.
Theriogenology. 2016 Mar 1;85(4):617-24. doi: 10.1016/j.theriogenology.2015.09.053. Epub 2015 Nov 14.
Cervical patency is considered to be important for uterine drainage after mating or artificial insemination (AI), and failure to relax or premature tightening of the cervix can lead to persistent endometritis. This study investigated the hypothesis that cervical occlusion after AI increases accumulation of fluid, polymorphonuclear leukocytes (PMNs), and cytokines in the uterine lumen. Endometrial swabs were obtained from 29 normal cyclic mares during the first, third, and fifth estrus and biopsies during the first and fifth estrus. All mares were inseminated during the second and fourth estrus. In either the second or fourth estrus, a clamped catheter was inserted into the uterus immediately after AI. Accumulation of intrauterine fluid was evaluated by transrectal ultrasonography at 0, 6, 25, and 48 hours. Fluid was drained from the catheter at either 25 hours (TxA) or 6 and 25 hours after AI (TxB). In the control estrus (TxC, no catheters), fluid was obtained by a tampon at 25 hours after AI. The uteri were then lavaged with Ringer's solution, after which the catheters were withdrawn. Sequences of treatments in the second and fourth estrus were A followed by C, C followed by A, B followed by C, and C followed by B in groups AC, CA, BC, and CB, respectively. Five mares lost their catheters and were excluded from the study. Scores for total inflammation, gland dilation, and lymphatic lacunae in the uterine biopsies did not differ significantly between groups or estrous periods. In contrast, periglandular fibrosis scores increased in all groups during the experiment. At 25 hours after AI in the second estrus, the mares with the catheters had larger accumulations of fluid (P < 0.05) and higher concentrations and total numbers of PMNs in uterine fluid (P < 0.05) than the mares without catheters. In the fourth estrus, the total number of PMNs was lower in TxB than in TxA at 25 hours (P < 0.05). Concentrations of PMNs in TxC were 10 times higher in the fourth estrus than the second. Within mare groups AC and BC, total numbers of PMNs in treatment C (fourth estrus) were as high as in TxA and B (second estrus). Expression of IL-1β, IL-6, IL-10 and TNF-α, analyzed by Western blotting, did not differ significantly between the treatments or estrous periods. It is concluded that a closed cervix after insemination results in pronounced inflammation of the mare's endometrium. Furthermore, this kind of severe insult may lead to permanent pathologic changes in the endometrium, including fibrosis.
子宫颈通畅被认为对交配或人工授精(AI)后的子宫引流很重要,子宫颈无法松弛或过早收紧会导致持续性子宫内膜炎。本研究调查了一个假设,即人工授精后子宫颈闭塞会增加子宫腔内液体、多形核白细胞(PMN)和细胞因子的积聚。在第1、3和5个发情期从29匹正常发情周期的母马获取子宫内膜拭子,并在第1和第5个发情期进行活检。所有母马在第2和第4个发情期进行人工授精。在第2或第4个发情期,人工授精后立即将一根夹紧的导管插入子宫。在0、6、25和48小时通过经直肠超声检查评估子宫内液体的积聚情况。在25小时(TxA)或人工授精后6和25小时(TxB)从导管排出液体。在对照发情期(TxC,无导管),在人工授精后25小时用棉塞获取液体。然后用林格氏液冲洗子宫,之后拔出导管。在第2和第4个发情期的治疗顺序在AC、CA、BC和CB组中分别为A后接C、C后接A、B后接C和C后接B。5匹母马的导管脱落,被排除在研究之外。子宫活检中总炎症、腺体扩张和淋巴腔隙的评分在各实验组或发情期之间没有显著差异。相比之下,在实验过程中所有组的腺周纤维化评分均增加。在第2个发情期人工授精后第25小时,有导管的母马比无导管的母马有更大的液体积聚(P<0.05),子宫液中PMN的浓度和总数更高(P<0.05)。在第4个发情期,TxB组在25小时时PMN的总数低于TxA组(P<0.05)。TxC组中第4个发情期PMN的浓度比第个发情期高10倍。在母马AC组和BC组内,治疗C组(第4个发情期)中PMN的总数与TxA组和TxB组(第2个发情期)一样高。通过蛋白质免疫印迹法分析,IL-1β、IL-6、IL-10和TNF-α的表达在各治疗组或发情期之间没有显著差异。得出的结论是,人工授精后子宫颈闭合会导致母马子宫内膜明显炎症。此外,这种严重损伤可能导致子宫内膜永久性病理变化,包括纤维化。