Ovarian steroid-dependent tumor necrosis factor-α production and its action on the equine endometrium in vitro.
Abstract: Tumor necrosis factor-α (TNF) is a cytokine that plays important roles in functions of the endometrium. The aims of this study were to determine whether (i) ovarian steroids modulate TNF production by endometrial cells (Experiment 1); (ii) TNF effects on prostaglandin (PG) production in cultured equine endometrial cells and tissue (Experiment 2). Epithelial and stromal cells were isolated from equine endometrium (Days 2-5 of the estrous cycle; n=20) and treated after passage 1. In Experiment 1, epithelial and stromal cells were exposed to progesterone (P4; 10(-7)M), 17-β estradiol (E2; 10(-9)M) or P4+E2 (10(-7)/10(-9)M) for 24h. Then, TNF mRNA transcription was determined using Real-time PCR. Additionally, TNF protein production was investigated in response to ovarian steroids for 24h using Enzyme-Linked Immunosorbent Spot (EliSpot). In Experiment 2, epithelial and stromal cells and endometrial explants (mid-luteal phase of the estrous cycle; n=5) were exposed in vitro to TNF (10 ng/ml) and to oxytocin (OT; positive control; 10(-7)M) for 24h. The concentrations of PGE2 and PGF2α were determined using a direct enzyme immunoassay (EIA) method. The transcription of prostaglandin-endoperoxide synthase-2 (PTGS-2), prostaglandin E2 synthase (PGES) and PGF2α synthase (PGFS) mRNAs in the endometrial explants was determined using Real-time PCR. Results showed that TNF is produced by two types of equine endometrial cells and its production is up-regulated by ovarian steroids (P<0.05) in stromal cells and by P4 (P<0.05) and E2 (P<0.01) in epithelial cells. Epithelial and stromal cells can also produce PG in response to TNF. In endometrial explants, TNF stimulated PGE2 production to a large extent and PGF2α secretion to a lesser extent. These actions are mediated by up-regulation of PG synthases mRNA transcription. The study indicates that TNF production is closely related to ovarian steroid actions and that the interaction between TNF and PG regulates physiologic processes in the equine endometrium.
Copyright © 2014 Elsevier Ltd. All rights reserved.
Publication Date: 2014-03-15 PubMed ID: 24642167DOI: 10.1016/j.cyto.2014.02.005Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study investigates the effects of ovarian steroids on the production of Tumor necrosis factor-α (TNF) in endometrial cells, as well as TNF’s impact on prostaglandin production in cultured equine endometrial cells and tissue.
Overview of the Study
- The researchers explored whether ovarian steroids modulate the production of TNF, a cytokine that plays crucial roles in the functioning of the endometrium. They also investigated how TNF influences the production of Prostaglandins (PG) in cultured equine endometrial cells and tissue.
- Two key experiments were conducted in this study. In the first experiment, endometrial cells (epithelial and stromal) isolated from equine endometrium were exposed to progesterone (P4) and 17-β estradiol (E2). The researchers then examined the levels of TNF mRNA transcription using Real-time PCR techniques.
- In the second experiment, endometrial explants and cells were exposed to TNF and oxytocin (OT) under in vitro conditions. The resultant quantities of PGE2 (Prostaglandin E2) and PGF2α (Prostaglandin F2 alpha) were then determined using a direct enzyme immunoassay (EIA) method.
Findings of the Study
- The findings demonstrated that TNF is produced by both types of endometrial cells (epithelial and stromal) and that its production is increased by ovarian steroids in stromal cells, and by P4 and E2 in epithelial cells.
- They also revealed that the exposure to TNF prompts both stromal and epithelial cells to produce Prostaglandins. In endometrial tissue samples, TNF highly stimulated the production of PGE2 and had a lesser effect on the secretion of PGF2α.
- The study concluded that TNF production and its action on the equine endometrium are tightly associated with ovarian steroid activity and that the interaction between TNF and Prostaglandins regulates physiological processes in the endometrium.
Significance of the Study
- The research provides new insights into the influence of ovarian hormones on the endometrium at cellular and molecular levels. Understanding the interaction between TNF and ovarian steroids could provide a basis for advanced fertility treatments and contribute to our understanding of reproductive biology.
- The results of this study are especially useful in veterinary science, mainly focusing on the equine reproduction system, as it advances our awareness of the intricate hormonal interactions and their consequences on equine fertility.
Cite This Article
APA
Szóstek AZ, Adamowski M, Galvão AM, Ferreira-Dias GM, Skarzynski DJ.
(2014).
Ovarian steroid-dependent tumor necrosis factor-α production and its action on the equine endometrium in vitro.
Cytokine, 67(2), 85-91.
https://doi.org/10.1016/j.cyto.2014.02.005 Publication
Researcher Affiliations
- Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research, Olsztyn, Poland.
- C.I.I.S.A., Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.
- Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research, Olsztyn, Poland; C.I.I.S.A., Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.
- C.I.I.S.A., Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.
- Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research, Olsztyn, Poland. Electronic address: d.skarzynski@pan.olsztyn.pl.
MeSH Terms
- Animals
- Cells, Cultured
- Cyclooxygenase 2 / genetics
- Dinoprost / metabolism
- Dinoprostone / metabolism
- Endometrium / cytology
- Endometrium / drug effects
- Endometrium / metabolism
- Enzyme-Linked Immunospot Assay
- Epithelial Cells / drug effects
- Epithelial Cells / metabolism
- Estradiol / pharmacology
- Female
- Gene Expression / drug effects
- Horses
- Hydroxyprostaglandin Dehydrogenases / genetics
- In Vitro Techniques
- Intramolecular Oxidoreductases / genetics
- Progesterone / pharmacology
- Prostaglandin-E Synthases
- Reverse Transcriptase Polymerase Chain Reaction
- Stromal Cells / drug effects
- Stromal Cells / metabolism
- Tumor Necrosis Factor-alpha / genetics
- Tumor Necrosis Factor-alpha / metabolism
- Tumor Necrosis Factor-alpha / pharmacology
Citations
This article has been cited 3 times.- Budik S, Walter I, Leitner MC, Ertl R, Aurich C. Expression of Enzymes Associated with Prostaglandin Synthesis in Equine Conceptuses.. Animals (Basel) 2021 Apr 20;11(4).
- Crociati M, Capomaccio S, Mandara MT, Stradaioli G, Sylla L, Monaci M, Cappelli K. Different expression of Defensin-B gene in the endometrium of mares of different age during the breeding season.. BMC Vet Res 2019 Dec 21;15(1):465.
- Siemieniuch MJ, Szóstek AZ, Gajos K, Kozdrowski R, Nowak M, Okuda K. Type of Inflammation Differentially Affects Expression of Interleukin 1β and 6, Tumor Necrosis Factor-α and Toll-Like Receptors in Subclinical Endometritis in Mares.. PLoS One 2016;11(5):e0154934.
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