Effects on the equine endometrium of cervical occlusion after insemination

Abstract

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 The uteri were then lavaged with Ringer's solution, after which the catheters were withdrawn.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.

Keywords: Cervix; Cytokine; Fibrosis; Inflammation; Intrauterine fluid; Mare.