[en] Myeloperoxidase (MPO), as a marker of neutrophil activation, has been associated with equine endometritis. However, in absence of inflammation, MPO is constantly detected in the uterine lumen of estrous mares. The aim of this study was to characterize MPO in the uterus of mares under physiological conditions as a first step to better understand the role of this enzyme in equine reproduction. Total and active MPO concentrations were determined, by ELISA and SIEFED assay, respectively, in low-volume lavages from mares in estrus (n = 26), diestrus (n = 18) and anestrus (n = 8) in absence of endometritis. Immunohistochemical analysis was performed on 21 endometrial biopsies randomly selected: estrus (n = 11), diestrus (n = 6) and anestrus (n = 4). MPO, although mostly enzymatically inactive, was present in highly variable concentrations in uterine lavages in all studied phases, with elevated concentrations in estrus and anestrus, while in diestrus, concentrations were much lower. Intracytoplasmic immunoexpression of MPO was detected in the endometrial epithelial cells, neutrophils and glandular secretions. Maximal expression was observed during estrus in mid and basal glands with a predominant intracytoplasmic apical reinforcement. In diestrus, immunopositive glands were sporadic. In anestrus, only the luminal epithelium showed residual MPO immunostaining. These results confirm a constant presence of MPO in the uterine lumen of mares in absence of inflammation, probably as part of the uterine mucosal immune system, and suggest that endometrial cells are a source of uterine MPO under physiological cyclic conditions.
Disciplines :
Veterinary medicine & animal health
Author, co-author :
Parrilla Hernandez, Sonia ; Université de Liège - ULiège > Fundamental and Applied Research for Animals and Health (FARAH)
Franck, Thierry ; Université de Liège - ULiège > Centres généraux > Centre de l'oxygène : Recherche et développement (C.O.R.D.)
Munaut, Carine ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques
Feyereisen, Emilie ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Biologie cellulaire et moléculaire
Piret, Joëlle ; Université de Liège - ULiège > Département de morphologie et pathologie (DMP)
Farnir, Frédéric ; Université de Liège - ULiège > Département de gestion vétérinaire des Ressources Animales (DRA)
Reigner, Fabrice; UE1297 PAO, INRAE, 37380 Nouzilly, France
Barrière, Philippe; UE1297 PAO, INRAE, 37380 Nouzilly, France
Deleuze, Stefan ; Université de Liège - ULiège > Fundamental and Applied Research for Animals and Health (FARAH) > FARAH: Médecine vétérinaire comparée
Language :
English
Title :
Characterization of Myeloperoxidase in the Healthy Equine Endometrium
Klebanoff S.J. Myeloperoxidase: Friend and foe J. Leukoc. Biol. 2005 77 598 625 10.1189/jlb.1204697 15689384
Borregaard N. Cowland J. Granules of the Human Neutrophilic Polymorphonuclear Leukocyte Blood 1997 89 3503 3521 10.1182/blood.V89.10.3503 9160655
Brinkmann V. Reichard U. Goosmann C. Fauler B. Uhlemann Y. Weiss D.S. Weinrauch Y. Zychlinsky A. Neutrophil Extra-cellular Traps Kill Bacteria Science 2004 303 1532 1535 10.1126/science.1092385 15001782
Khan A.A. Alsahli M.A. Rahmani A.H. Myeloperoxidase as an Active Disease Biomarker: Recent Biochemical and Pathological Perspectives Med. Sci. 2018 6 33 10.3390/medsci6020033
Friedrichs K. Baldus S. Klinke A. Fibrosis in Atrial Fibrillation—Role of Reactive Species and MPO Front. Physiol. 2012 3 214 10.3389/fphys.2012.00214
Rudolph V. Andrié R.P. Rudolph T.K. Friedrichs K. Klinke A. Hirsch-Hoffmann B. Schwoerer A.P. Lau D. Fu X. Klingel K. et al. Myeloperoxidase acts as a profibrotic mediator of atrial fibrillation Nat. Med. 2010 16 470 474 10.1038/nm.2124
Pulli B. Ali M. Iwamoto Y. Zeller M.W. Schob S. Linnoila J.J. Chen J.W. Myeloperoxidase–Hepatocyte–Stellate Cell Cross Talk Promotes Hepatocyte Injury and Fibrosis in Experimental Nonalcoholic Steatohepatitis Antioxid. Redox Signal. 2015 23 1255 1269 10.1089/ars.2014.6108
Austin G.E. Zhao W.-G. Adjiri A. Lu J.-P. Control of myeloperoxidase gene expression in developing myeloid cells Leuk. Res. 1996 20 817 820 10.1016/S0145-2126(96)00032-X
Green P.S. Mendez A.J. Jacob J.S. Crowley J.R. Growdon W. Hyman B.T. Heinecke J.W. Neuronal expression of myeloperoxidase is increased in Alzheimer’s disease J. Neurochem. 2004 90 724 733 10.1111/j.1471-4159.2004.02527.x
Martínez-Marcos P. Carvajal-Serna M. Lázaro-Gaspar S. Pérez-Pé R. Muiño-Blanco T. Cebrián-Pérez J.A. Casao A. Presence of melatonin-catabolizing non-specific enzymes myeloperoxidase and indoleamine 2,3-dioxygenase in the ram reproductive tract Reprod Dom. Anim. 2019 54 1643 1650 10.1111/rda.13574
Roumeguère T. Delree P. Van Antwerpen P. Rorive S. Vanhamme L. Ryhove L.D.L.K.D. Serteyn D. Wespes E. Vanhaerverbeek M. Boudjeltia K.Z. Intriguing location of myeloperoxidase in the prostate: A preliminary immunohistochemical study Prostate 2011 72 507 513 10.1002/pros.21452
Yang Z. Xiao K. Wang W. Tang J. Sun P.-P. Peng K.-M. Song H. The Effect of Visfatin on Inflammatory Reaction in Uterus of LPS-Induced Rats Int. J. Morphol. 2015 33 194 203 10.4067/S0717-95022015000100031
Haegens A. Vernooy J.H.J. Heeringa P. Mossman B.T. Wouters E.F.M. Myeloperoxidase modulates lung epithelial responses to pro-inflammatory agents Eur. Respir. J. 2008 31 252 260 10.1183/09031936.00029307
Yang J.J. Preston G.A. Pendergraft W.F. Segelmark M. Heeringa P. Hogan S.L. Jennette J.C. Falk R.J. Internalization of Proteinase 3 Is Concomitant with Endothelial Cell Apoptosis and Internalization of Myeloperoxidase with Generation of Intracellular Oxidants Am. J. Pathol. 2001 158 581 592 10.1016/S0002-9440(10)64000-X
Jerke U. Rolle S. Purfürst B. Luft F. Nauseef W. Kettritz R. β2 Integrin-mediated Cell-Cell Contact Transfers Active Myeloperoxidase from Neutrophils to Endothelial Cells* J. Biol. Chem. 2013 288 12910 12919 10.1074/jbc.M112.434613
La Rocca G. Di Stefano A. Eleuteri E. Anzalone R. Magno F. Corrao S. Loria T. Martorana A. Di Gangi C. Colombo M. et al. Oxidative stress induces myeloperoxidase expression in endocardial endothelial cells from patients with chronic heart failure Basic Res. Cardiol. 2009 104 307 320 10.1007/s00395-008-0761-9
Art T. Franck T. Lekeux P. de Moffarts B. Couëtil L. Becker M. Kohnen S. Deby-Dupont G. Serteyn D. Myeloperoxidase con-centration in bronchoalveolar lavage fluid from healthy horses and those with recurrent airway obstruction Can. J. Vet. Res. 2006 70 291 296
Wauters J. Pille F. Martens A. Franck T. Serteyn D. Gasthuys F. Meyer E. Equine myeloperoxidase: A novel biomarker in synovial fluid for the diagnosis of infection Equine Vet. J. 2012 45 278 283 10.1111/j.2042-3306.2012.00682.x
Ponthier J. Franck T. Parrilla-Hernandez S. Niesten A. De La Rebiere G. Serteyn D. Deleuze S. Concentration, Activity and Biochemical Characterization of Myeloperoxidase in Fresh and Post-Thaw Equine Semen and their Implication on Freezability Reprod. Domest. Anim. 2014 49 285 291 10.1111/rda.12270
Art T. Franck T. Gangl M. Votion D. Kohnen S. Deby-Dupont G. Serteyn D. Plasma concentrations of myeloperoxidase in endurance and 3-day event horses after a competition Equine Vet. J. 2006 38 298 302 10.1111/j.2042-3306.2006.tb05557.x 17402436
Grulke S. Franck T. Gangl M. Péters F. Salciccia A. Deby-Dupont G. Serteyn D. Myeloperoxidase assay in plasma and peri-toneal fluid of horses with gastrointestinal disease Can. J. Vet. Res. 2008 72 37 42 18214160
Storms N. Torres C.M. Franck T. Guitart A.S. de la Rebière G. Serteyn D. Presence of Myeloperoxidase in Lamellar Tissue of Horses Induced by an Euglycemic Hyperinsulinemic Clamp Front. Vet. Sci. 2022 9 6835 10.3389/fvets.2022.846835 35359667
Parrilla-Hernandez S. Ponthier J. Franck T.Y. Serteyn D.D. Deleuze S.C. High concentrations of myeloperoxidase in the equine uterus as an indicator of endometritis Theriogenology 2014 81 936 940 10.1016/j.theriogenology.2014.01.011
Rebordão M.R. Amaral A. Fernandes C. Silva E. Lukasik K. Szóstek-Mioduchowska A. Pinto-Bravo P. Galvão A. Skarzynski D.J. Ferreira-Dias G. Enzymes Present in Neutrophil Extracellular Traps May Stimulate the Fibrogenic PGF2α Pathway in the Mare Endometrium Animals 2021 11 2615 10.3390/ani11092615
Rebordão M.R. Amaral A. Lukasik K. Szóstek-Mioduchowska A. Pinto-Bravo P. Galvão A. Skarzynski D.J. Ferreira-Dias G. Constituents of neutrophil extracellular traps induce in vitro collagen formation in mare endometrium Theriogenology 2018 113 8 18 10.1016/j.theriogenology.2018.02.001
Rebordão M. Carneiro C. Alexandre-Pires G. Brito P. Pereira C. Nunes T. Galvão A. Leitão A. Vilela C. Ferreira-Dias G. Neutrophil extracellular traps formation by bacteria causing endometritis in the mare J. Reprod. Immunol. 2014 106 41 49 10.1016/j.jri.2014.08.003
Nazhat S.A. Kitahara G. Kozuka N. Mido S. Sadawy M. Ali H.E.-S. Osawa T. Associations of periparturient plasma biochemical parameters, endometrial leukocyte esterase and myeloperoxidase, and bacterial detection with clinical and subclinical en-dometritis in postpartum dairy cows J. Vet. Med. Sci. 2018 80 302 310 10.1292/jvms.17-0478
Amaral A. Fernandes C. Rebordão M. Szóstek-Mioduchowska A. Lukasik K. Pinto-Bravo P. da Gama L.T. Skarzynski D.J. Ferreira-Dias G. Myeloperoxidase Inhibition Decreases the Expression of Collagen and Metallopeptidase in Mare Endometria under In Vitro Conditions Animals 2021 11 208 10.3390/ani11010208
Chapman A.L.P. Mocatta T.J. Shiva S. Seidel A. Chen B. Khalilova I. Paumann-Page M.E. Jameson G.N.L. Winterbourn C.C. Kettle A.J. Ceruloplasmin Is an Endogenous Inhibitor of Myeloperoxidase J. Biol. Chem. 2013 288 6465 6477 10.1074/jbc.M112.418970
Segelmark M. Persson B. Hellmark T. Wieslander J. Binding and inhibition of myeloperoxidase (MPO): A major function of ceruloplasmin? Clin. Exp. Immunol. 1997 108 167 174 10.1046/j.1365-2249.1997.d01-992.x
Panzani D. Di Vita M. Lainé A.-L. Guillaume D. Rota A. Tesi M. Vannozzi I. Camillo F. Corpus Luteum Vascularization and Progesterone Production in Autumn and Winter Cycles of the Mare: Relationship Between Ultrasonographic Characteristics of Corpora Lutea and Plasma Progesterone Concentration in the Last Cycles Before Anestrus J. Equine Vet. Sci. 2017 56 35 39 10.1016/j.jevs.2017.05.001
LeBlanc M.M. Magsig J. Stromberg A.J. Use of a low-volume uterine flush for diagnosing endometritis in chronically infertile mares Theriogenology 2007 68 403 412 10.1016/j.theriogenology.2007.04.038
Kozdrowski R. Sikora M. Buczkowska J. Nowak M. Raś A. Dzięcioł M. Effects of cycle stage and sampling procedure on interpretation of endometrial cytology in mares Anim. Reprod. Sci. 2015 154 56 62 10.1016/j.anireprosci.2015.01.009
Overbeck W. Witte T. Heuwieser W. Comparison of three diagnostic methods to identify subclinical endometritis in mares Theriogenology 2011 75 1311 1318 10.1016/j.theriogenology.2010.12.002
Kenney R.M.D.P. Equine Endometrial Biopsy Current Therapy in Theriogenology Da M. W.B Saunders Company Philadelphia, PA, USA 1986 723 729
Schoon H.-A. Schoon D. Klug E. Endometrial biopsies as an ancillary aid in diagnosis and prognosis of subfertility in the mare Pferdeheilkunde Equine Med. 1992 8 355 362 10.21836/PEM19920603
Franck T. Grulke S. Deby-Dupont G. Deby C. Duvivier H. Péters F. Serteyn D. Development of an enzyme-linked im-munosorbent assay for specific equine neutrophil myeloperoxidase measurement in blood J. Vet. Diagn. Invest. 2005 17 412 419 10.1177/104063870501700502
Franck T. Kohnen S. Deby-Dupont G. Grulke S. Deby C. Serteyn D. A Specific Method for Measurement of Equine Active Myeloperoxidase in Biological Samples and in in Vitro Tests J. Vet. Diagn. Investig. 2006 18 326 334 10.1177/104063870601800402
Hoffmann C. Bazer F.W. Klug J. Aupperle H. Ellenberger C. Schoon H.A. Immunohistochemical and histochemical identifi-cation of proteins and carbohydrates in the equine endometrium Theriogenology 2009 71 264 274 10.1016/j.theriogenology.2008.07.008
Khalilova I.S. Dickerhof N. Mocatta T.J. Bhagra C.J. McClean D.R. Obinger C. Kettle A.J. A myeloperoxidase precursor, pro-myeloperoxidase, is present in human plasma and elevated in cardiovascular disease patients PLoS ONE 2018 13 e0192952 10.1371/journal.pone.0192952
Kenney R. Doig P. Equine endometrial biopsy Curr. Ther. Ther. 1986 2 723 729
Hoffmann C. Bazer F.W. Klug J. Aupperle H. Ellenberger C. Özgen S. Schoon D. Schoon H.-A. Immunohistochemical and histochemical identification of proteins and carbohydrates in the equine endometrium: Reaction patterns in the cycling mare Pferdeheilkunde Equine Med. 2009 25 212 219 10.21836/PEM20090303
Schöniger S. Gräfe H. Richter F. Schoon H.-A. Expression of indoleamine 2,3-dioxygenase 1 as transcript and protein in the healthy and diseased equine endometrium Res. Vet. Sci. 2018 118 278 287 10.1016/j.rvsc.2018.03.001 29547726
Schöniger S. Gräfe H. Schoon H.-A. Beta-defensin is a component of the endometrial immune defence in the mare Pferdeheilkunde Equine Med. 2013 29 335 346 10.21836/PEM20130307
Schöniger S. Böttcher D. Theuß T. Schoon H.-A. Expression of Toll-like receptors 2, 4 and 6 in equine endometrial epithelial cells: A comparative in situ and in vitro study Res. Vet. Sci. 2017 112 34 41 10.1016/j.rvsc.2017.01.004
Churg A. Anderson W.A. Induction of Endometrial Peroxidase Synthesis and Secretion by Estrogen and Estrogen Antagonist J. Cell Biol. 1974 62 449 459 10.1083/jcb.62.2.449
McRae A.C. The blood-uterine lumen barrier and exchange between extracellular fluids Reproduction 1988 82 857 873 10.1530/jrf.0.0820857
Ceusters J.D. Mouithys-Mickalad A.A. Franck T.J. Derochette S. Vanderplasschen A. Deby-Dupont G.P. Serteyn D.A. Effect of myeloperoxidase and anoxia/reoxygenation on mitochondrial respiratory function of cultured primary equine skeletal myoblasts Mitochondrion 2013 13 410 416 10.1016/j.mito.2012.12.004
de Araujo T.H. Okada S.S. Ghosn E.E. Taniwaki N.N. Rodrigues M.R. de Almeida S.R. Mortara R.A. Russo M. Campa A. Albu-querque R.C. Intracellular localization of myeloperoxidase in murine peritoneal B-lymphocytes and macrophages Cell Immunol. 2013 281 27 30 10.1016/j.cellimm.2013.01.002
Wira C.R. Grant-Tschudy K.S. Crane-Godreau M.A. Epithelial Cells in the Female Reproductive Tract: A Central Role as Sentinels of Immune Protection Am. J. Reprod. Immunol. 2005 53 65 76 10.1111/j.1600-0897.2004.00248.x
Kolm G. Klein D. Knapp E. Watanabe K. Walter I. Lactoferrin expression in the horse endometrium: Relevance in persisting mating-induced endometritis Veter- Immunol. Immunopathol. 2006 114 159 167 10.1016/j.vetimm.2006.08.005
Katila T. Lock T. Hoffmann W. Smith A. Lysozyme, alkaline phosphatase and neutrophils in uterine secretions of mares with differing resistance to endometritis Theriogenology 1990 33 723 732 10.1016/0093-691X(90)90549-9 16726768
Shibata T. Sakamoto J. Osaka Y. Neyatani N. Fujita S. Oka Y. Takagi H. Mori H. Fujita H. Tanaka Y. et al. Myelop-eroxidase in blood neutrophils during normal and abnormal menstrual cycles in women of reproductive age Int. J. Lab. Hematol. 2017 39 169 174 10.1111/ijlh.12599
Békési G. Kakucs R. Varbiro S. Feher J. Pazmany T. Magyar Z. Sprintz D. Szekacs B. Induced myeloperoxidase activity and related superoxide inhibition during hormone replacement therapy BJOG Int. J. Obstet. Gynaecol. 2001 108 474 481 10.1111/j.1471-0528.2001.00108.x
Houdeau E. Larauche M. Monnerie R. Bueno L. Fioramonti J. Uterine motor alterations and estrous cycle disturbances as-sociated with colonic inflammation in the rat Am. J. Physiol. Regul. Integr. Comp. Physiol. 2005 288 630 637 10.1152/ajpregu.00482.2004
Baiza-Gutman L. Flores-Sánchez M. Diaz-Flores M. Hicks J. Presence of uterine peroxidase activity in the rat early pregnancy Int. J. Biochem. Cell Biol. 2000 32 255 262 10.1016/S1357-2725(99)00061-8
Cummings A.M. Metcalf J.L. Mechanisms of the stimulation of rat uterine peroxidase activity by methoxychlor Reprod. Toxicol. 1994 8 477 486 10.1016/0890-6238(94)90030-2
Farley D.B. Ford S.P. Rosazza J.P.N. Increase in uterine peroxidase activity in the rat uterus during oestrogen hyperaemia Reproduction 1992 95 551 558 10.1530/jrf.0.0950551
Okada H. Nawa M. Kimura J. In Vitro Effect of Estrogens on the Peroxidase Activity of Human Endometrium* J. Clin. Endocrinol. Metab. 1985 61 404 407 10.1210/jcem-61-3-404
Batra S.C. Iosif C.S. Effect of Estrogen Treatment on the Peroxidase Activity and Estrogen Receptors in the Female Rabbit Urogenital Tissues J. Urol. 1992 148 935 938 10.1016/S0022-5347(17)36781-2
Grant-Tschudy K.S. Wira C.R. Effect of Estradiol on Mouse Uterine Epithelial Cell Transepithelial Resistance (TER) Am. J. Reprod. Immunol. 2004 52 252 262 10.1111/j.1600-0897.2004.00218.x 15494046
Lewis G.S. Role of ovarian progesterone and potential role of prostaglandin F2α and prostaglandin E2 in modulating the uterine response to infectious bacteria in postpartum ewes J. Anim. Sci. 2003 81 285 293 10.2527/2003.811285x 12597400
Watson E.D. Stokes C.R. Bourne F.J. Influence of administration of ovarian steroids on the function of neutrophils isolated from the blood and uterus of ovariectomized mares J. Endocrinol. 1987 112 443 448 10.1677/joe.0.1120443
Chadwick J.W. Fine N. Khoury W. Tasevski N. Sun C. Boroumand P. Klip A. Glogauer M. Tissue-specific murine neutrophil activation states in health and inflammation J. Leukoc. Biol. 2020 110 187 195 10.1002/JLB.4AB1020-248RRR 33145850
Seals R.C. Wulster-Radcliffe M.C. Lewis G.S. Modulation of the Uterine Response to Infectious Bacteria in Postpartum Ewes Am. J. Reprod. Immunol. 2002 47 57 63 10.1034/j.1600-0897.2002.1o062.x
Klebanoff S.J. Iodination of Bacteria: A Bactericidal Mechanism J. Exp. Med. 1967 126 1063 1078 10.1084/jem.126.6.1063
Davies M.J. Hawkins C.L. The Role of Myeloperoxidase in Biomolecule Modification, Chronic Inflammation, and Disease Antioxid. Redox Signal. 2020 32 957 981 10.1089/ars.2020.8030
Kargapolova Y. Geißen S. Zheng R. Baldus S. Winkels H. Adam M. The Enzymatic and Non-Enzymatic Function of Myeloperoxidase (MPO) in Inflammatory Communication Antioxidants 2021 10 562 10.3390/antiox10040562
Reilas T. del Alamo M.M.R. Liepina E. Yeste M. Katila T. Effects on the equine endometrium of cervical occlusion after insemination Theriogenology 2016 85 617 624 10.1016/j.theriogenology.2015.09.053
Ricketts S.W. Alonso S. The effect of age and parity on the development of equine chronic endometrial disease Equine Vet. J. 1991 23 189 192 10.1111/j.2042-3306.1991.tb02752.x
Bradley P.P. Christensen R.D. Rothstein G. Cellular and Extracellular Myeloperoxidase in Pyogenic Inflammation Blood 1982 60 618 622 10.1182/blood.V60.3.618.618
Edwards S.W. Nurcombe H.L. A Hart C. Oxidative inactivation of myeloperoxidase released from human neutrophils Biochem. J. 1987 245 925 928 10.1042/bj2450925
King C.C. Jefferson M.M. Thomas E.L. Secretion and inactivation of myeloperoxidase by isolated neutrophils J. Leukoc. Biol. 1997 61 293 302 10.1002/jlb.61.3.293
Klebanoff S.J. Effect of estrogens on the myeloperoxidase-mediated antimicrobial system Infect. Immun. 1979 25 153 156 10.1128/iai.25.1.153-156.1979
Galijasevic S. Abdulhamid I. Abu-Soud H.M. Melatonin Is a Potent Inhibitor for Myeloperoxidase Biochemistry 2008 47 2668 2677 10.1021/bi702016q
Galijasevic S. Abdulhamid I. Abu-Soud H.M. Potential role of tryptophan and chloride in the inhibition of human myelop-eroxidase Free Radic. Biol. 2008 44 1570 1577 10.1016/j.freeradbiomed.2008.01.003
Eklinke A. Nussbaum C. Kubala L. Friedrichs K. Rudolph T.K. Rudolph V. Paust H.-J. Schröder C. Benten D. Lau D. et al. Myeloperoxidase attracts neutrophils by physical forces Blood 2011 117 1350 1358 10.1182/blood-2010-05-284513
Lefkowitz D.L. Gelderman M.P. Fuhrmann S.R. Graham S. Starnes J.D. Lefkowitz S.S. Bollen A. Moguilevsky N. Neutrophilic Myeloperoxidase–Macrophage Interactions Perpetuate Chronic Inflammation Associated with Experimental Arthritis Clin. Immunol. 1999 91 145 155 10.1006/clim.1999.4696
Lefkowitz D.L. Roberts E. Grattendick K. Schwab C. Stuart R. Lincoln J. Allen R.C. Moguilevsky N. Bollen A. Lefkowitz S.S. The Endothelium and Cytokine Secretion: The Role of Peroxidases as Immunoregulators Cell Immunol. 2000 202 23 30 10.1006/cimm.2000.1638