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Adiponectin and Chemerin in the endometrium of postpartum dairy cows with cytological endometritis
Pereira, G.; Bexiga, R.; Silva, J.C...
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Objective: To evaluate the relationship between the gene transcription and expression of Adiponectin (ADIPOQ, ADIPOR1 and ADIPOR2) and Chemerin (RARRES2 and CM- KLR1), and the inflammatory status of the uterus of postpartum dairy cows.
Materials and methods: High-yielding postpartum dairy cows (n=36) without puerperal disease were retrospectively allocated to groups: i) Healthy (H; n=6), without cytological endometritis (CE) at 25 and 45 days postpartum (dpp) and pregnant at first AI; ii) CE-H (n=19), with CE at 25 dpp, but that recovered by 45 dpp; iii) CE-CE (n=11), with persistent CE until 45 dpp. Endometrial cytology was assessed from uterine swabs taken with a cytobrush device at 25 and 45 dpp (cut-off values: 18 % and 5 %, respectively). At 45 dpp, a low volume lavage followed by centrifugation, allowed to obtain cellular pellet and supernatant samples of each uterine horn. An endometrial biopsy was also taken from each uterine horn. Gene transcription of ADIPOQ and its receptors (ADIPOR1 and ADIPOR2), and of chemerin (RARRES2) and its receptor (CMKLR1) was analysed in the cellular pellet by quantitative real-time PCR. Protein expression was analysed by immunohistochemistry (IHC) in endometrial biopsy samples and protein production (ADIPOQ and RARRES2) by ELISA in the supernatants.
Results: Transcription levels of ADIPOQ and ADIPOR2 were higher (p < 0.001) in CE-CE than in H and CE-H cows, whereas ADIPOR1 mRNA levels were lower (p < 0.05) in CE- CE cows than in H cows. Transcription levels of RARRES2 and CMKLR1 were higher in CE-CE than in H and CE-H cows (p < 0.01). Positive immunostaining for ADIPOQ was observed in the luminal and glandular epithelium, endothelial cells, stro- ma and inflammatory cells of all cows. However, CE-CE cows exhibited a stronger staining than H and CE-H cows. Staining for ADIPOR1 was only observed in the luminal and glandular epithelium, whereas ADIPOR2 also stained in stroma and in- flammatory cells. Staining for RARRES2 and CMKLR1 was observed in all endometrial compartments of all cows. However, RARRES2 staining showed a stronger signal in CE-CE than in H and CE-H cows. Uterine fluid ADIPOQ and RAR- RES2 concentrations were higher (p < 0.001) in CE-CE than in H and CE-H cows.
Conclusions: At 45 dpp, cows with persistent CE show up-regulated gene transcription and protein expression of Adiponectin and Chemerin in the endometrium, compared with healthy cows and cows that recovered from CE. Results support a local production of these mediators, and a relationship with the inflammatory status of the postpartum uterus of dairy cows. The local production and signalling of these adipokines prompt for an autocrine and/or paracrine role in the inflammatory response exhibited by cows with subclinical endometritis. This turns these adipokines into potential biomarkers of endometrial inflammation, namely to identify cows at risk of persistent subclinical endometritis. However, further studies are warranted to clarify their role in the establishment of subclinical endometritis.
Funding: FORMAS (Grant No 2015-00888) and CIISA UIDP/CVT/00276/2020. Gonçalo Pereira has a PhD grant from FCT (SFRH/BD/130923/2017).
Keywords: Adiponectin, Chemerin, Endometritis, Cow.
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Affiliation of the authors at the time of publication
CIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de Medicina Veterinária, Universidade de Lisboa, Lisboa, Portugal;
INRAE, UMR85 Physiologie de la Reproduction et des Comportements, 37380, Nouzilly, France;
Division of Reproduction, Department of Clinical Sciences, SLU, Uppsala, Sweden.
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