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Frontiers in veterinary science2020; 7; 631; doi: 10.3389/fvets.2020.00631

Collagen and Eosinophils in Jenny’s Endometrium: Do They Differ With Endometrial Classification?

Abstract: Collagen fibers and inflammatory cells are the basis for jenny endometrium Kenney and Doig's classification developed for the mare. The infiltration of a large number of eosinophils in the jenny endometrium is intriguing. Eosinophil and fibroblast produced IL33, which has been related to fibrosis development and chronicity. This work on the endometrium consisted of (i) quantification of collagen type I (COL1A2), type III (COL3A1), and IL33 transcripts; (ii) histological localization and quantification of COL1 and COL3 proteins; and (iii) eosinophil and neutrophil count and correlation with collagen area and IL33 transcripts. Localization of COL protein in the jenny endometrium was also compared to the mare endometrium. As fibrosis increased, eosinophil and neutrophil count decreased ( < 0.05). A 5-fold increase in IL33 transcripts was noted from categories IIA to III. There was a tendency toward a positive correlation between eosinophil count and IL33 transcripts in category IIA endometrium ( = 0.055). Neither transcripts of COL1A2 nor COL3A1 nor the areas of COL1 or COL3 differed with endometrial categories. Unlike for the mare, and regardless of the jenny endometrium classification, COL3 was always found to different extents in the stratum compactum, while COL1 was mainly present in deep stroma. As fibrosis progressed in the mare, an extensive increase in COL1 fibers was notorious under the surface epithelium. Correlations between neutrophil count and COL1 and COL3 areas were observed in the jenny endometrium, although no correlation was found for eosinophil count. Neutrophil count positive correlation with the COL1 area and negative correlation with the COL3 area in endometria with mild lesions suggest that neutrophils in the jenny endometrium may be involved in fibrogenesis. In addition, when eosinophilia subsides, the endometrium reacts with fibrosis establishment, which could be stimulated by the pro-fibrotic cytokine IL33, whose release might then be ascribed to fibroblasts. Further studies are needed to analyze the effect of the presence of COL3 next to the surface epithelium in the stratum compactum, or around the endometrial glands on jenny's endometrial function and fertility.
Publication Date: 2020-09-10 PubMed ID: 33134338PubMed Central: PMC7511575DOI: 10.3389/fvets.2020.00631Google Scholar: Lookup
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  • Journal Article

Summary

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This research investigates the relationship between different types of collagen, inflammatory cells like eosinophils and neutrophils, and interleukin 33 (IL33) in the endometrium of jennies (female donkeys), by comparing it to mares (female horses). The finding suggests that neutrophils may contribute to fibrosis and eosinophils may stimulate the release of IL33, a pro-fibrotic cytokine.

Study objectives and method

  • The overarching goal of this study was to understand better the role of collagen, eosinophils, and IL33 in Jenny endometrium. Jenny endometrium refers to the lining of the uterus in female donkeys, called jennies.
  • The researchers used several methods to achieve this goal. They quantified the presence of collagen type I and type III (known as COL1A2 and COL3A1 respectively) and IL33, a protein associated with immune response and inflammation.
  • They also noted the number of eosinophils and neutrophils, types of white blood cells that are key to the body’s immune response, and looked for any correlations between these cell types and collagen or IL33.

Key findings

  • As fibrotic tissue (scar tissue caused by fibrosis) increased, the study observed a decrease in the count of both eosinophils and neutrophils.
  • Interestingly, a 5-fold increase in the protein IL33 was found between certain categories of fibrosis. There seemed to be a possible correlation between the presence of eosinophils and IL33 in category IIA endometrium.
  • The location of collagen proteins in jenny endometrium differed from mare endometrium (female horse endometrium). While collagen type III was present at different extents in all classifications of jenny endometrium, collagen type I was found mainly in the deeper layers.
  • And although no correlations were found with eosinophil count, correlations were observed between neutrophil count and COL1 and COL3 areas, suggesting that neutrophils could play a role in fibrogenesis (the formation of fibrous tissue).

Takeaway and further studies

  • The findings suggest that as eosinophilia (high eosinophil levels) subsides, the endometrium reacts by developing fibrosis. This action could be stimulated by IL33 and could mean that fibroblasts (a type of cell that makes the extracellular matrix and collagen) are responsible for its release.
  • There are still more studies needed to further understand the effects of collagen type III’s presence near the epithelium surface and the endometrial glands of jenny’s endometrium on fertility and overall endometrial function.

Cite This Article

APA
Miró J, Gutiérrez-Reinoso M, da Silva JA, Fernandes C, Rebordão MR, Alexandre-Pires G, Catalán J, Ferreira-Dias G. (2020). Collagen and Eosinophils in Jenny’s Endometrium: Do They Differ With Endometrial Classification? Front Vet Sci, 7, 631. https://doi.org/10.3389/fvets.2020.00631

Publication

ISSN: 2297-1769
NlmUniqueID: 101666658
Country: Switzerland
Language: English
Volume: 7
Pages: 631
PII: 631

Researcher Affiliations

Miró, Jordi
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Cerdanyola del Vallès, Spain.
Gutiérrez-Reinoso, Miguel
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Cerdanyola del Vallès, Spain.
da Silva, Joana Aguiar
  • Faculdade de Medicina Veterinária, CIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Universidade de Lisboa, Lisbon, Portugal.
Fernandes, Carina
  • Faculdade de Medicina Veterinária, CIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Universidade de Lisboa, Lisbon, Portugal.
Rebordão, Maria Rosa
  • Faculdade de Medicina Veterinária, CIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Universidade de Lisboa, Lisbon, Portugal.
  • Coimbra College of Agriculture, Polytechnic Institute of Coimbra, Coimbra, Portugal.
Alexandre-Pires, Graça
  • Faculdade de Medicina Veterinária, CIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Universidade de Lisboa, Lisbon, Portugal.
Catalán, Jaime
  • Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, Cerdanyola del Vallès, Spain.
Ferreira-Dias, Graça
  • Faculdade de Medicina Veterinária, CIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Universidade de Lisboa, Lisbon, Portugal.

References

This article includes 41 references
  1. Vilés K, Rabanal R, Rodríguez-Prado M, Miró J. Effect of ketoprofen treatment on the uterine inflammatory response after AI of jennies with frozen semen.. Theriogenology 2013 Apr 15;79(7):1019-26.
  2. Rothenberg ME, Hogan SP. The eosinophil.. Annu Rev Immunol 2006;24:147-74.
  3. Gervasi MG, Osycka-Salut C, Caballero J, Vazquez-Levin M, Pereyra E, Billi S, Franchi A, Perez-Martinez S. Anandamide capacitates bull spermatozoa through CB1 and TRPV1 activation.. PLoS One 2011 Feb 11;6(2):e16993.
  4. McHedlidze T, Waldner M, Zopf S, Walker J, Rankin AL, Schuchmann M, Voehringer D, McKenzie AN, Neurath MF, Pflanz S, Wirtz S. Interleukin-33-dependent innate lymphoid cells mediate hepatic fibrosis.. Immunity 2013 Aug 22;39(2):357-71.
  5. Rankin AL, Mumm JB, Murphy E, Turner S, Yu N, McClanahan TK, Bourne PA, Pierce RH, Kastelein R, Pflanz S. IL-33 induces IL-13-dependent cutaneous fibrosis.. J Immunol 2010 Feb 1;184(3):1526-35.
    doi: 10.4049/jimmunol.0903306pubmed: 20042577google scholar: lookup
  6. Masterson JC, Capocelli KE, Hosford L, Biette K, McNamee EN, de Zoeten EF, Harris R, Fernando SD, Jedlicka P, Protheroe C, Lee JJ, Furuta GT. Eosinophils and IL-33 Perpetuate Chronic Inflammation and Fibrosis in a Pediatric Population with Stricturing Crohn's Ileitis.. Inflamm Bowel Dis 2015 Oct;21(10):2429-40.
  7. Kenney RM. The aetiology, diagnosis and classification of chronic degenerative endometritis (CDE) (endometriosis). In: Allen WR, editor. Equine Endometritis Worship. Proceedings of the John P. Hughes International Workshop on Equine Endometritis; 1992 Aug 19-20. Newmarket (1993).
  8. Hoffmann C, Ellenberger C, Mattos RC, Aupperle H, Dhein S, Stief B, Schoon HA. The equine endometrosis: new insights into the pathogenesis.. Anim Reprod Sci 2009 Apr;111(2-4):261-78.
  9. Bochsler PN, Slauson DO. Inflammation and repair of tissue. In: Slauson DO, Cooper BJ, editors. Mechanisms of Disease: A Textbook of Comparative General Pathology. St. Louis, MO: Mosby; (2002). p. 140–245.
  10. Pinto-Bravo P, Reborddo MR, Amoral A, Fernandes C, Cuello C, Parrilla I. Is mare endometrosis linked to oviduct fibrosis?. Pferdeheilkunde (2018) 34:43–6.
    doi: 10.21836/PEM20180107google scholar: lookup
  11. Junqueira LC, Cossermelli W, Brentani R. Differential staining of collagens type I, II and III by Sirius Red and polarization microscopy.. Arch Histol Jpn 1978 Jun;41(3):267-74.
    doi: 10.1679/aohc1950.41.267pubmed: 82432google scholar: lookup
  12. Podico G, Canisso IF, Roady PJ, Austin SM, Carossino M, Balasuriya U, Ellerbrock RE, Lima FS, Ferreira-Dias G, Douglas RH. Uterine responses and equine chorionic gonadotropin concentrations after two intrauterine infusions with kerosene post early fetal loss in mares.. Theriogenology 2020 Apr 15;147:202-210.
  13. Porto CD. Caracterização histoquímica do colágeno e expressão de MMP-2, MMP-9 e TIMP-1 nas endometrites crônicas das éguas. (Dissertation/Master's thesis). Botucatu: University Estadual Paulista “Júlio de Mesquita Filho”; (2006).
  14. Kenney R, Doig PA. Equine endometrial biopsy. In: Morrow DA, editor. Current Therapy in Theriogenology. Philadelphia, PA: WB Saunders; (1986). p. 723–9.
  15. Rebordão MR, Amaral A, Lukasik K, Szóstek-Mioduchowska A, Pinto-Bravo P, Galvão A, Skarzynski DJ, Ferreira-Dias G. Impairment of the antifibrotic prostaglandin E(2) pathway may influence neutrophil extracellular traps-induced fibrosis in the mare endometrium.. Domest Anim Endocrinol 2019 Apr;67:1-10.
  16. Dheda K, Huggett JF, Bustin SA, Johnson MA, Rook G, Zumla A. Validation of housekeeping genes for normalizing RNA expression in real-time PCR.. Biotechniques 2004 Jul;37(1):112-4, 116, 118-9.
    doi: 10.2144/04371RR03pubmed: 15283208google scholar: lookup
  17. Chrysanthopoulou A, Mitroulis I, Apostolidou E, Arelaki S, Mikroulis D, Konstantinidis T, Sivridis E, Koffa M, Giatromanolaki A, Boumpas DT, Ritis K, Kambas K. Neutrophil extracellular traps promote differentiation and function of fibroblasts.. J Pathol 2014 Jul;233(3):294-307.
    doi: 10.1002/path.4359pubmed: 24740698google scholar: lookup
  18. Zhao S, Fernald RD. Comprehensive algorithm for quantitative real-time polymerase chain reaction.. J Comput Biol 2005 Oct;12(8):1047-64.
    doi: 10.1089/cmb.2005.12.1047pmc: PMC2716216pubmed: 16241897google scholar: lookup
  19. Gao Q, Li Y, Li M. The potential role of IL-33/ST2 signaling in fibrotic diseases.. J Leukoc Biol 2015 Jul;98(1):15-22.
    doi: 10.1189/jlb.3RU0115-012Rpubmed: 25881899google scholar: lookup
  20. Millar NL, O'Donnell C, McInnes IB, Brint E. Wounds that heal and wounds that don't - The role of the IL-33/ST2 pathway in tissue repair and tumorigenesis.. Semin Cell Dev Biol 2017 Jan;61:41-50.
    doi: 10.1016/j.semcdb.2016.08.007pubmed: 27521518google scholar: lookup
  21. Liew FY. IL-33: a Janus cytokine.. Ann Rheum Dis 2012 Apr;71 Suppl 2:i101-4.
  22. Chan BCL, Lam CWK, Tam LS, Wong CK. IL33: Roles in Allergic Inflammation and Therapeutic Perspectives.. Front Immunol 2019;10:364.
    doi: 10.3389/fimmu.2019.00364pmc: PMC6409346pubmed: 30886621google scholar: lookup
  23. Santulli P, Borghese B, Chouzenoux S, Vaiman D, Borderie D, Streuli I, Goffinet F, de Ziegler D, Weill B, Batteux F, Chapron C. Serum and peritoneal interleukin-33 levels are elevated in deeply infiltrating endometriosis.. Hum Reprod 2012 Jul;27(7):2001-9.
    doi: 10.1093/humrep/des154pubmed: 22587998google scholar: lookup
  24. Miller JE, Monsanto SP, Ahn SH, Khalaj K, Fazleabas AT, Young SL, Lessey BA, Koti M, Tayade C. Interleukin-33 modulates inflammation in endometriosis.. Sci Rep 2017 Dec 20;7(1):17903.
    doi: 10.1038/s41598-017-18224-xpmc: PMC5738435pubmed: 29263351google scholar: lookup
  25. Bulun SE. Endometriosis.. N Engl J Med 2009 Jan 15;360(3):268-79.
    doi: 10.1056/NEJMra0804690pubmed: 19144942google scholar: lookup
  26. Itoga T, Matsumoto T, Takeuchi H, Yamasaki S, Sasahara N, Hoshi T, Kinoshita K. Fibrosis and smooth muscle metaplasia in rectovaginal endometriosis.. Pathol Int 2003 Jun;53(6):371-5.
  27. Vicino M, Scioscia M, Resta L, Marzullo A, Ceci O, Selvaggi LE. Fibrotic tissue in the endometrioma capsule: surgical and physiopathologic considerations from histologic findings.. Fertil Steril 2009 Apr;91(4 Suppl):1326-8.
  28. Sokkar SM, Hamouda MA, El-Rahman SM. Endometritis in she donkeys in Egypt.. J Vet Med B Infect Dis Vet Public Health 2001 Sep;48(7):529-36.
  29. Canisso IF, Panzani D, Miró J, Ellerbrock RE. Key Aspects of Donkey and Mule Reproduction.. Vet Clin North Am Equine Pract 2019 Dec;35(3):607-642.
    doi: 10.1016/j.cveq.2019.08.014pubmed: 31672204google scholar: lookup
  30. Akuthota P, Weller PF. Eosinophils and disease pathogenesis.. Semin Hematol 2012 Apr;49(2):113-9.
  31. Grimm AL, Schoon HA, Schöniger S. Histopathological features of endometritis eosinophilica in mares.. Histol Histopathol 2017 Nov;32(11):1161-1173.
    doi: 10.14670/HH-11-872pubmed: 28105630google scholar: lookup
  32. Zeisberg M, Kalluri R. Cellular mechanisms of tissue fibrosis. 1. Common and organ-specific mechanisms associated with tissue fibrosis.. Am J Physiol Cell Physiol 2013 Feb 1;304(3):C216-25.
    doi: 10.1152/ajpcell.00328.2012pmc: PMC3566435pubmed: 23255577google scholar: lookup
  33. Sziksz E, Pap D, Lippai R, Béres NJ, Fekete A, Szabó AJ, Vannay Á. Fibrosis Related Inflammatory Mediators: Role of the IL-10 Cytokine Family.. Mediators Inflamm 2015;2015:764641.
    doi: 10.1155/2015/764641pmc: PMC4495231pubmed: 26199463google scholar: lookup
  34. Pires MA, Ferreira-Dias G, Catarino J, Bastos-de-Carvalho C, Silvestre M, Nóvoa M. Correlation between morphological characterization of jenny (Equus asinus) endometrial biopsy and quantification of collagen deposition by image analysis. J Comp Pathol (2019) 166:139.
  35. Masseno APB. Assessment of endometrial fibrosis and myofibroblasts in endometriosis active and inactive mares. (Dissertation/Ph.D. thesis). Botucatu: University Estadual Paulista “Júlio de Mesquita Filho”; (2012).
  36. Costa LD. Histochemical and immunohistochemical characterization of fibrotic changes in mares endometrosis. (Dissertation/Master's thesis). Botucatu: University Estadual Paulista “Júlio de Mesquita Filho”; (2015).
  37. Kenney RM. Cyclic and pathologic changes of the mare endometrium as detected by biopsy, with a note on early embryonic death.. J Am Vet Med Assoc 1978 Feb 1;172(3):241-62.
    pubmed: 621166
  38. Walter I, Handler J, Reifinger M, Aurich C. Association of endometriosis in horses with differentiation of periglandular myofibroblasts and changes of extracellular matrix proteins.. Reproduction 2001 Apr;121(4):581-6.
    doi: 10.1530/reprod/121.4.581pubmed: 11277878google scholar: lookup
  39. Rebordão MR, Amaral A, Lukasik K, Szóstek-Mioduchowska A, Pinto-Bravo P, Galvão A, Skarzynski DJ, Ferreira-Dias G. Constituents of neutrophil extracellular traps induce in vitro collagen formation in mare endometrium.. Theriogenology 2018 Jun;113:8-18.
  40. Reilas T, Rivera Del Alamo MM, Liepina E, Yeste M, Katila T. Effects on the equine endometrium of cervical occlusion after insemination.. Theriogenology 2016 Mar 1;85(4):617-24.
  41. Quartuccio M, Cristarella S, Medica P, Fazio E, Mazzullo G, Rifici C, Liotta L, Satué K. Endometrial Cytology During the Different Phases of the Estrous Cycle in Jennies: New Evidences.. Animals (Basel) 2020 Jun 19;10(6).
    doi: 10.3390/ani10061062pmc: PMC7341222pubmed: 32575538google scholar: lookup

Citations

This article has been cited 6 times.
  1. Alpoim-Moreira J, Fernandes C, Pimenta J, Bliebernicht M, Rebordão MR, Castelo-Branco P, Szóstek-Mioduchowska A, Skarzynski DJ, Ferreira-Dias G. Metallopeptidades 2 and 9 genes epigenetically modulate equine endometrial fibrosis. Front Vet Sci 2022;9:970003.
    doi: 10.3389/fvets.2022.970003pubmed: 36032279google scholar: lookup
  2. 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).
    doi: 10.3390/ani11102918pubmed: 34679937google scholar: lookup
  3. Yánez-Ortiz I, Catalán J, Mateo-Otero Y, Dordas-Perpinyà M, Gacem S, Yeste N, Bassols A, Yeste M, Miró J. Extracellular Reactive Oxygen Species (ROS) Production in Fresh Donkey Sperm Exposed to Reductive Stress, Oxidative Stress and NETosis. Antioxidants (Basel) 2021 Aug 27;10(9).
    doi: 10.3390/antiox10091367pubmed: 34572999google scholar: lookup
  4. Vallejo-Soto P, Dorado J, Herrera-García R, Álvarez-Delgado C, Gómez-Laguna J, Santiago Á, Manrique M, González Ariza A, León Jurado JM, Hidalgo M, Ortiz I. Histomorphometric Analysis of the Endometrium of Jennies (Equus asinus) and Mares (Equus caballus) in Estrus: Anatomical Differences and Possible Reproductive Implications. Animals (Basel) 2026 Jan 4;16(1).
    doi: 10.3390/ani16010143pubmed: 41514829google scholar: lookup
  5. Fazaeli H, Davoodi F, Sheikholeslami A, Sheykhhasan M, Kalhor N, Naserpour L, Jannatifar R, Sahraei SS. Simultaneous effect of naringenin and beta-catenin signaling inhibitor C-82 on modulating gene expression and functional pattern of mesenchymal stem cells from endometriosis patients. Iran J Basic Med Sci 2025;28(5):671-679.
  6. Wang Z, Zeng S, Wang Y, Wang T, Qu H, Zhu Y, Li J. Reproductive Disorders in Donkeys: Current Evidence and Update. Animals (Basel) 2024 Aug 28;14(17).
    doi: 10.3390/ani14172494pubmed: 39272279google scholar: lookup