Endometrial IL-1beta, IL-6 and TNF-alpha, mRNA expression in mares resistant or susceptible to post-breeding endometritis. Effects of estrous cycle, artificial insemination and immunomodulation.
Abstract: Endometrial mRNA expression of the pro-inflammatory cytokines interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-alpha) was assessed in mares resistant (RM) or susceptible (SM) to persistent post-breeding endometritis (PPBE). Eight RM and eight SM, were selected based on reproductive records and functional tests out of a herd of 2,000 light cross-type mares. Three experiments were done to study transcription patterns in (i) basal conditions; (ii) after artificial insemination (AI); and (iii) after administration of an immunomodulator at time of artificial insemination. Endometrial biopsies were taken during consecutive cycles: (i) at estrus, when follicles reached 35 mm and at diestrus (7 +/- 1 days after ovulation); (ii) at 24 h post-AI, with dead semen (estrus) and in diestrus; (iii) at 24 h after treatment with a Mycobacterium phlei cell-wall extract (MCWE) preparation and AI (with dead semen), and at diestrus. mRNA expression was quantitated by real time PCR. Under basal conditions, SM had significantly higher mRNA expression of all cytokines in estrus and of IL-1beta and TNF-alpha in diestrus, compared to RM. After AI, there were no differences between RM and SM in estrus; however, mRNA expression for all three pro-inflammatory cytokines was higher than under basal conditions. In diestrus, RM showed significantly lower IL-1beta and TNF-alpha mRNA expression than SM. When MCWE was administered at time of AI, no differences between cytokine induction from RM and SM were found. Globally, mRNA expression for all three cytokines correlated well among themselves when expression was high. The present study showed that (i) in basal conditions RM had lower mRNA expression of pro-inflammatory cytokines than SM with no effect of estrous cycle; (ii) AI upregulated mRNA expression for all three cytokines in both RM and SM, with persistance in diestrus in the latter; (iii) treatment with MCWE at time of AI down-regulated mRNA expression of IL-1 with significant effects in SM which behaved like RM. Immunomodulation with MCWE could be of help in restoring homeostatic local inflammatory mechanisms, thus assisting in the prophylaxis of post-breeding endometritis in mares.
Publication Date: 2003-10-03 PubMed ID: 14522132DOI: 10.1016/s0165-2427(03)00137-5Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article explores the expression of certain inflammatory substances in horses that are either resistant or susceptible to a type of uterine inflammation following breeding. The study also looked at how this expression changes under normal conditions, after artificial insemination, and following treatment with an immune system altering substance.
Objective and Sample Selection
- In this study, the researchers wanted to assess the endometrial expression of certain inflammatory substances (pro-inflammatory cytokines), namely interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-alpha), in mares that are either resistant to or susceptible to persistent post-breeding endometritis, a type of uterine inflammation that can occur after breeding.
- For the study, the researchers selected eight resistant mares (RM) and eight susceptible mares (SM) from a herd of 2,000 light cross-type mares, with selection based on their reproductive records and functional tests.
Experiments and Measurements
- The researchers conducted three experiments to examine transcription patterns in different conditions: under normal circumstances, after artificial insemination, and after administration of an immunomodulator (a substance that adjusts immune response) called Mycobacterium phlei cell-wall extract (MCWE) at the time of artificial insemination.
- Biopsies were taken from the mares during consecutive cycles to enable measurement of the mRNA (messenger RNA) expressions of the cytokines under these different conditions.
- The amount of mRNA expressions was measured using real-time PCR, a method used for amplifying and simultaneously quantifying a targeted DNA molecule.
Key Findings
- Under regular circumstances, SM had significantly higher mRNA expressions of all the cytokines in estrus (the period of sexual receptivity in mares) and higher expression of IL-1beta and TNF-alpha in diestrus (the period of sexual non-receptivity), compared to RM. This suggests that SM produce more of the inflammation-causing cytokines, which could make them more prone to post-breeding endometritis.
- After artificial insemination, mRNA expressions for all three cytokines increased in both RM and SM, with this increase being more persistent in SM. RM showed lower cytokine levels in diestrus compared to SM. This may indicate that artificial insemination, and possibly breeding in general, makes mares more prone to inflammation.
- Administration of MCWE at the time of insemination lowered cytokine production, particularly in SM, which behaved similarly to RM after treatment. This suggests that immunomodulation with MCWE could help balance local inflammatory reactions, thus potentially preventing post-breeding endometritis in mares.
Cite This Article
APA
Fumuso E, Giguère S, Wade J, Rogan D, Videla-Dorna I, Bowden RA.
(2003).
Endometrial IL-1beta, IL-6 and TNF-alpha, mRNA expression in mares resistant or susceptible to post-breeding endometritis. Effects of estrous cycle, artificial insemination and immunomodulation.
Vet Immunol Immunopathol, 96(1-2), 31-41.
https://doi.org/10.1016/s0165-2427(03)00137-5 Publication
Researcher Affiliations
- Departamento de Clínica, Facultad de Ciencias Veterinarias, UNICEN, Pinto 399, 7000 Tandil, Argentina.
MeSH Terms
- Adjuvants, Immunologic / pharmacology
- Animals
- Biopsy / veterinary
- Disease Susceptibility
- Endometritis / immunology
- Endometritis / veterinary
- Estrous Cycle / immunology
- Female
- Horse Diseases / immunology
- Horses
- Insemination, Artificial / immunology
- Insemination, Artificial / veterinary
- Interleukin-1 / biosynthesis
- Interleukin-1 / genetics
- Interleukin-6 / biosynthesis
- Interleukin-6 / genetics
- Male
- Mycobacterium / immunology
- Pregnancy
- RNA, Messenger / biosynthesis
- RNA, Messenger / genetics
- Reverse Transcriptase Polymerase Chain Reaction / veterinary
- Tumor Necrosis Factor-alpha / biosynthesis
- Tumor Necrosis Factor-alpha / genetics
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