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Elastase inhibition affects collagen transcription and prostaglandin secretion in mare endometrium during the estrous cycle.
Abstract: We have shown that bacteria induce neutrophil extracellular traps (NETs) in mare endometrium. Besides killing pathogens, NETs may contribute for endometrosis (chronic endometrium fibrosis). Since elastase (ELA) is a NETs component that regulates fibrosis and prostaglandin (PG) output, the aim was to evaluate if inhibition of ELA would affect collagen 1 (COL1) transcription and PGs secretion by endometrium explants, in different estrous cycle phases. Follicular-FP (n = 8) and mid luteal-MLP (n = 7) phases explants were cultured for 24-48 hr with medium alone (Control), ELA (0.5 μg/ml,1 μg/ml), sivelestat - ELA inhibitor (INH,10 μg/ml), or ELA (0.5 μg/ml,1 μg/ml) + INH (10 μg/ml). COL1 gene transcription was done by qRT-PCR and PGE2 and PGF2 α determination in culture medium by EIA. In FP, at 24 hr, ELA0.5 increased COL1 transcription (p < 0.001) but its inhibition (ELA0.5 + INH10) decreased COL1 transcription (p < 0.01) and PGF2 α production (p < 0.05). Also, ELA0.5 + INH10 or ELA1 + INH10 raised PGE2 production (p < 0.01). At 48 hr, ELA1 increased COL1 transcription (p < 0.01) and PGF2 α production (p < 0.001), but its inhibition (ELA1 + INH10) decreased these actions (p < 0.01; p < 0.05, respectively). Besides, ELA1 + INH10 incubation increased PGE2 (p < 0.05). PGF2 α also augmented with ELA0.5 (p < 0.001), but lowered with ELA0.5 + INH10 (p < 0.01). In MLP, ELA0.5 up-regulated COL1 transcription (24 hr, p < 0.01; 48 hr, p < 0.001), but ELA0.5 + INH10 decreased it (24 hr, p < 0.05; 48 hr, p < 0.001). At 48 hr, incubation with ELA1 also increased COL1 transcription and PGF2 α production (p < 0.05), but PGF2 α production decreased with ELA1 + INH10 incubation (p < 0.05). PGE2 production was higher in ELA1 + INH10 incubation (p < 0.05). Therefore, ELA inhibition may reduce the establishment of mare endometrial fibrosis by stimulating the production of anti-fibrotic PGE2 and inhibiting pro-fibrotic PGF2 α.
© 2018 Blackwell Verlag GmbH.
Publication Date: 2018-09-22 PubMed ID: 30238664DOI: 10.1111/rda.13258Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research investigates how restricting a compound called elastase (ELA), which contributes to fibrosis (scarring) in the endometrium (uterine lining) of mares, affects the production of collagen and prostaglandins, substances linked to inflammation and pain. The study found that inhibiting ELA production could potentially decrease mare endometrial fibrosis.
Research Methodology
The team conducted an experiment by culturing endometrium explants taken from mares in different estrous cycle stages: Follicular-FP and mid luteal-MLP. The explants were cultured for a period of 24-48 hours using three main input variations:
- Medium alone, serving as the control group.
- ELA (0.5 μg/ml,1 μg/ml)
- Sivelestat – which is an ELA inhibitor (INH,10 μg/ml)
- ELA (0.5 μg/ml,1 μg/ml) + INH (10 μg/ml)
Using qRT-PCR, they monitored the transcription of collagen 1 (COL1), a protein responsible for tissue structuring that’s prevalent in fibrotic tissue. They also measured the levels of prostaglandin E (PGE) and prostaglandin F alpha (PGF α), substances that affect inflammation and wound healing, in the culture medium by EIA.
Main Findings
The researchers found that ELA and its inhibition had significant effects on the transcription of COL1 and the production of PGF α and PGE, with these effects varying across the estrous cycle phases and time.
- In the FP phase, 24-hour incubation with 0.5 μg/ml ELA increased COL1 transcription, but this action was mitigated when ELA was inhibited. Augmented PGE production was also noticed when ELA was inhibited.
- Furthermore, at 48 hours, 1 μg/ml ELA increased COL1 transcription and PGF α production; however, these increases were curtailed with ELA inhibition.
- In the MLP phase, low concentration ELA (0.5 μg/ml) up-regulated COL1 transcription both at 24-hour and 48-hour intervals, but this transcription was decreased when ELA was inhibited.
- While a higher concentration of ELA (1 μg/ml) increased COL1 transcription and PGF α production at 48 hours, these results were diminished with ELA inhibition.
Conclusion and Implication
The research concludes that inhibiting ELA could reduce the occurrence of endometrial fibrosis in mares. This occurs by stimulating the production of PGE, known for its anti-fibrotic properties, and inhibiting pro-fibrotic PGF α. This knowledge could potentially be used to develop therapeutic interventions for endometrial fibrosis in mares.
Cite This Article
APA
Amaral A, Fernandes C, Lukasik K, Szóstek-Mioduchowska A, Baclawska A, Rebordão MR, Aguiar-Silva J, Pinto-Bravo P, Skarzynski DJ, Ferreira-Dias G.
(2018).
Elastase inhibition affects collagen transcription and prostaglandin secretion in mare endometrium during the estrous cycle.
Reprod Domest Anim, 53 Suppl 2, 66-69.
https://doi.org/10.1111/rda.13258 Publication
Researcher Affiliations
- CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.
- Institute of Animal Reproduction and Food Research, Polish Academy of Science, Olsztyn, Poland.
- CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.
- Institute of Animal Reproduction and Food Research, Polish Academy of Science, Olsztyn, Poland.
- Institute of Animal Reproduction and Food Research, Polish Academy of Science, Olsztyn, Poland.
- Institute of Animal Reproduction and Food Research, Polish Academy of Science, Olsztyn, Poland.
- CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.
- Polytechnic Institute of Coimbra, Coimbra College of Agriculture, Coimbra, Portugal.
- CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.
- CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.
- Polytechnic Institute of Coimbra, Coimbra College of Agriculture, Coimbra, Portugal.
- Institute of Animal Reproduction and Food Research, Polish Academy of Science, Olsztyn, Poland.
- CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary Medicine, University of Lisbon, Lisbon, Portugal.
MeSH Terms
- Animals
- Collagen / genetics
- Collagen / metabolism
- Dinoprost / metabolism
- Dinoprostone / metabolism
- Endometrium / drug effects
- Estrous Cycle
- Female
- Horses / physiology
- Pancreatic Elastase / pharmacology
Citations
This article has been cited 12 times.- Ferreira-Dias GM, Alpoim-Moreira J, Szóstek-Mioduchowska A, Rebordão MR, Skarzynski DJ. The path to fertility: Current approaches to mare endometritis and endometrosis. Anim Reprod 2024;21(3):e20240070.
- Zdrojkowski Ł, Pawliński B, Skierbiszewska K, Jasiński T, Domino M. Assessment of Connective Tissue in the Equine Uterus and Cervix: Review of Clinical Impact and Staining Options. Animals (Basel) 2024 Jan 3;14(1).
- Amaral A, Cebola N, Szóstek-Mioduchowska A, Rebordão MR, Kordowitzki P, Skarzynski D, Ferreira-Dias G. Inhibition of Myeloperoxidase Pro-Fibrotic Effect by Noscapine in Equine Endometrium. Int J Mol Sci 2023 Feb 10;24(4).
- Katila T, Ferreira-Dias G. Evolution of the Concepts of Endometrosis, Post Breeding Endometritis, and Susceptibility of Mares. Animals (Basel) 2022 Mar 19;12(6).
- Amaral A, Fernandes C, Szóstek-Mioduchowska A, Lukasik K, Rebordão MR, Pinto-Bravo P, Skarzynski DJ, Ferreira-Dias G. The Inhibitory Effect of Noscapine on the In Vitro Cathepsin G-Induced Collagen Expression in Equine Endometrium. Life (Basel) 2021 Oct 19;11(10).
- Fantini P, Jiménez R, Vilés K, Iborra A, Palhares MS, Catalán J, Prades M, Miró J. Simple Tube Centrifugation Method for Platelet-Rich Plasma (PRP) Preparation in Catalonian Donkeys as a Treatment of Endometritis-Endometrosis. Animals (Basel) 2021 Oct 9;11(10).
- Rebordão MR, Amaral A, Fernandes C, Silva E, Lukasik K, Szóstek-Mioduchowska A, Pinto-Bravo P, Galvão A, Skarzynski DJ, Ferreira-Dias G. Enzymes Present in Neutrophil Extracellular Traps May Stimulate the Fibrogenic PGF(2α) Pathway in the Mare Endometrium. Animals (Basel) 2021 Sep 6;11(9).
- Amaral A, Fernandes C, Szóstek-Mioduchowska A, Rebordão MR, Skarzynski DJ, Ferreira-Dias G. Noscapine Acts as a Protease Inhibitor of In Vitro Elastase-Induced Collagen Deposition in Equine Endometrium. Int J Mol Sci 2021 May 19;22(10).
- Amaral A, Fernandes C, Rebordão MR, Szóstek-Mioduchowska A, Lukasik K, Pinto-Bravo P, Telo da Gama L, Jan Skarzynski D, Ferreira-Dias G. Myeloperoxidase Inhibition Decreases the Expression of Collagen and Metallopeptidase in Mare Endometria under In Vitro Conditions. Animals (Basel) 2021 Jan 16;11(1).
- Amaral A, Fernandes C, Morazzo S, Rebordão MR, Szóstek-Mioduchowska A, Lukasik K, Gawronska-Kozak B, Telo da Gama L, Skarzynski DJ, Ferreira-Dias G. The Inhibition of Cathepsin G on Endometrial Explants With Endometrosis in the Mare. Front Vet Sci 2020;7:582211.
- Amaral A, Fernandes C, Rebordão MR, Szóstek-Mioduchowska A, Lukasik K, Gawronska-Kozak B, Telo da Gama L, Skarzynski DJ, Ferreira-Dias G. The In Vitro Inhibitory Effect of Sivelestat on Elastase Induced Collagen and Metallopeptidase Expression in Equine Endometrium. Animals (Basel) 2020 May 16;10(5).
- Canisso IF, Segabinazzi LGTM, Fedorka CE. Persistent Breeding-Induced Endometritis in Mares - a Multifaceted Challenge: From Clinical Aspects to Immunopathogenesis and Pathobiology. Int J Mol Sci 2020 Feb 20;21(4).
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