Abstract:The aim of the study was to characterize endometrial mRNA transcription, immunolocalization, and protein expression of interleukin (IL) 1alpha, IL1beta, IL6, and IL1RI, IL1RII, and IL6Ralpha/beta in the course of endometrosis during the estrous cycle. Additionally, the influence of IL1alpha, IL1beta, and IL6 on prostaglandin (PG) secretion and PG synthase mRNA transcription in endometrial tissue during endometrosis was investigated. The endometrial samples were obtained at the early (n = 12), mid- (n = 12), an… Show more
“…The sequences for equine IL-1RI , IL-1RII , IL-6R β , and ACTB primers were previously published [23]. After a preliminary study, ACTB was chosen as the best housekeeping gene.…”
The aim of the present study was to investigate the interaction between ovarian steroids, interleukins and prostaglandins (PG) in equine epithelial and stromal cells in vitro. In Experiment 1, cells were exposed to IL-1α (10 ng/mL), IL-1β (10 ng/mL) or IL-6 (10 ng/mL) for 24 h and cell proliferation was determined using MTT. In Experiment 2, cells were exposed to progesterone (P4; 10−7 M); 17-β estradiol (E2; 10−9 M) or P4+E2 for 24 h and later medium was replaced with a fresh one treated with IL-1α, IL-1β or IL-6 (10 ng/mL, each) for 24 h. The oxytocin (OT; 10−7 M) was used as a positive control. In Experiment 3, cells were exposed to P4 (10−7 M), E2 (10−9 M) or P4+E2 for 24 h and the IL receptor mRNAs transcription was determined using Real-time PCR. Prostaglandins concentration was determined using the direct enzyme immunoassay (EIA) method. Our findings reveal a functional linking between ovarian steroids and IL-stimulated PG secretion by equine endometrial cells. This interaction could be one of the mechanisms responsible for endometrial local orchestrating events during the estrous cycle and early pregnancy.
“…The sequences for equine IL-1RI , IL-1RII , IL-6R β , and ACTB primers were previously published [23]. After a preliminary study, ACTB was chosen as the best housekeeping gene.…”
The aim of the present study was to investigate the interaction between ovarian steroids, interleukins and prostaglandins (PG) in equine epithelial and stromal cells in vitro. In Experiment 1, cells were exposed to IL-1α (10 ng/mL), IL-1β (10 ng/mL) or IL-6 (10 ng/mL) for 24 h and cell proliferation was determined using MTT. In Experiment 2, cells were exposed to progesterone (P4; 10−7 M); 17-β estradiol (E2; 10−9 M) or P4+E2 for 24 h and later medium was replaced with a fresh one treated with IL-1α, IL-1β or IL-6 (10 ng/mL, each) for 24 h. The oxytocin (OT; 10−7 M) was used as a positive control. In Experiment 3, cells were exposed to P4 (10−7 M), E2 (10−9 M) or P4+E2 for 24 h and the IL receptor mRNAs transcription was determined using Real-time PCR. Prostaglandins concentration was determined using the direct enzyme immunoassay (EIA) method. Our findings reveal a functional linking between ovarian steroids and IL-stimulated PG secretion by equine endometrial cells. This interaction could be one of the mechanisms responsible for endometrial local orchestrating events during the estrous cycle and early pregnancy.
“…; Szóstek et al. ). Changes noted in the expression of interleukins (IL)1‐ α , IL‐1 β , IL‐6 and their receptors in endometrium with different degrees of fibrosis and/or inflammation, might be oestrous‐cycle hormone dependent (Szóstek et al.…”
Section: Physiopathologic Mechanisms Of Endometrosismentioning
confidence: 97%
“…Changes noted in the expression of interleukins (IL)1‐ α , IL‐1 β , IL‐6 and their receptors in endometrium with different degrees of fibrosis and/or inflammation, might be oestrous‐cycle hormone dependent (Szóstek et al. ). However, early after breeding, a lower mRNA expression of IL‐6 and IL‐10 was detected in susceptible to persistent breeding‐induced endometritis mares, in which endometrosis animals were included (Woodward et al.…”
Section: Physiopathologic Mechanisms Of Endometrosismentioning
confidence: 99%
“…Also in the equine endometrium, we have shown that in the course of endometrosis, there are changes in PG production and mRNA transcription of prostaglandin ( PG ) synthases (Szóstek et al. , ). We hypothesize that prostanoid modifications might be involved in oestrous‐cycle disorders, early embryo mortality and subsequent infertility.…”
Section: Physiopathologic Mechanisms Of Endometrosismentioning
Contents
Endometrosis is a degenerative chronic process, characterized by paramount fibrosis development in mare endometrium. This condition is one of the major causes of subfertility/infertility in mares. As in other organs, fibrosis might be a pathologic sequel of many chronic inflammatory diseases. However, aetiology and physiopathologic mechanisms involved in endometrial fibrosis are still controversial. This review presents new hypotheses based on our newest data. As the first line of innate immune defence, systemic neutrophils arrive in the uterus at mating or in the presence of pathogens. A novel paradigm is that neutrophils cast out their DNA in response to infectious stimuli and form neutrophil extracellular traps (NETs). We have shown that bacterial strains of Streptococcus zooepidemicus, Escherichia coli or Staphylococcus capitis, known to cause endometritis in mares were able to induce NETs release in vitro by equine PMN to different extents. An intriguing dilemma is the dual action of NETs. While NETs play a desirable role fighting micro‐organisms in mare uterus, they may also contribute to endometrial fibrosis. A long‐term in vitro exposure of mare endometrium explants to NETs components (myeloperoxidase, elastase and cathepsin G) up‐regulated fibrosis markers TGFβ and Tissue inhibitor of metalloproteinase (TIMP‐1). Also, pro‐fibrotic cytokines regulated collagen deposition and fibrosis. Changes in expression of connective tissue growth factor (CTGF), interleukins (IL)1‐α, IL‐1β, IL‐6 and receptors in endometrium with different degrees of fibrosis and/or inflammation were observed. A putative role of CTGF, IL and NETs components in endometrosis development should be considered. Additionally, we speculate that in sustained endometritis in mares, prostaglandins may not only cause early luteolysis or early pregnancy loss, but may also be related to endometrial fibrosis pathogenesis by stimulating collagen deposition.
“…On the other hand, cytokines play an important role in the reproductive processes [39]. The complexity of their regulation is due to pleiotropism of cytokines, where each cytokine has multiple target cells of different organs and where responses may differ according to cell type.…”
In cyclic mares, the uterine environment can easily disturbed due to inflammatory processes that occur secondary to microbial invasion. Different aerobic and anaerobic bacteria can enter the uterus during natural mating, artificial insemination, reproductive examination or parturition. The postpartum period is a critical phase since due to relaxation of the uterus and cervix may favor recurrent infections air intake pneumovagina or urine collection in mares with poor perineal conformation. Infections are mainly caused by opportunistic or commensal microorganisms, such as Streptococcus zooepidemicus, hemolytic Escherichia coli and Staphylococcus aureus. Other microorganisms like Taylorella equigenitalis, Klebsiella pneumoniae and Pseudomona aeruginosa are transmitted through venereal route. Regarding to the fungal endometritis, the most common fungi include Candida and Aspergillus. These microorganisms cause infertility as a result of repeated inseminations during the breeding season and proliferate when the natural immune system is weakened in mares with advanced age and multiparous, or after repeated use of antibiotics. Indeed, in susceptible mare to endometritis, uterine defense mechanisms involving phagocytosis and opsonization by neutrophils, local synthesis of antibodies, mucociliary activity, vascular and myoelectric activity permeability are compromised, leading to fluid accumulation in response to inflammation and infertility.
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