Expression profiling of selected miRNAs in equine endometrium in response to LPS challenge in vitro: A new understanding of the inflammatory immune response.
Abstract: Bacterial infections of the genital tract are the major cause of reproductive failure in the mares. MiRNAs are important regulators of gene expression, mostly through transcriptional and translational regression. We hypothesized that LPS induced aberrant expression of miRNAs and their targets, which are involved in regulation of uterine homeostasis. Three groups of primary endometrial epithelial and stromal cells, and endometrial tissue explants were cultured. The 1 group was kept as control, while the 2 and 3 groups were challenged with low (0.5 μg/mL) or high (3.0 μg/mL) doses of Lipopolysaccharides (LPS). Cell pellets and tissue explants were collected after 24 and 48 h, for total RNA isolation and qRT-PCR of the selected miRNAs and their targets. Culture media and cell lysates were collected after 24 and 48 h, for cytokines (IL6 and TNF) and prostaglandins (PGE & PGF) measurement. Both endometrial cells expressed TLR4 and its accessory molecules (MyD88 & CD14) that are required for triggering inflammatory immune response after LPS, via up-regulation of TRAF6, TNF, IL6 and IL8, compared to the respective control. After both doses of LPS challenge, miR-155, miR-223 and miR-17 were significantly increased; miR-181b, miR-21 and let-7a were significantly decreased compared to respective controls. Interestingly, miR-24 and miR-532-5p were clearly up-regulated after only the low LPS dose. TNF, IL6 and PGs in culture media and from cell lysates revealed dose- and time-dependent patterns, after LPS. Results indicated that both epithelial and stromal cells have a primary role in innate immune response after LPS challenge, while this recognition occurred via TLR4 and its accessory molecules. Dysregulation of miRNAs and their targets expression after LPS might affect normal uterine function through perturbation of PG and cytokine secretion.
Copyright © 2019 Elsevier B.V. All rights reserved.
Publication Date: 2019-02-16 PubMed ID: 30885304DOI: 10.1016/j.vetimm.2019.02.006Google Scholar: Lookup
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- Journal Article
Summary
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The research studies the effect of bacterial infection on reproductive failures in mares by examining how Lipopolysaccharides (LPS) affect the expression of miRNAs (microRNAs) and their target genes. The results suggest that the dysregulation of miRNAs might interfere with normal uterine function through disturbance of prostaglandin and cytokine secretion.
Introduction and Research Background
- The study focuses on bacterial infections in the genital tract, which are a major cause of reproductive failure in mares.
- MicroRNAs (miRNAs), which regulate gene expression, are hypothesized to undergo aberrant expression in response to LPS, a component of bacteria that triggers an inflammatory response.
- This aberrant expression of miRNAs possibly disrupts the regular functioning of the uterus.
Experimental Design and Method
- In the experiment, three groups of primary endometrial cells and tissue were cultured, with one group serving as the control.
- The other two groups were treated with low and high doses of LPS for comparison.
- After 24 and 48 hours, cell and tissue samples were collected for RNA isolation and analysis of miRNAs and their targets.
- Cytokines and prostaglandins in the culture media and cell lysates were also measured at these times.
Results and Findings
- Both kinds of endometrial cells (epithelial and stromal) expressed TLR4 (Toll-like receptor 4) and its accessory molecules (MyD88 & CD14), which are necessary for triggering an inflammatory response after exposure to LPS.
- After exposure to LPS, several miRNAs exhibited significant changes in their expression compared to the control.
- Importantly, miR-24 and miR-532-5p showed increased expression only with the low dose of LPS.
- cytokines (TNF, IL6) and prostaglandins in the culture media and from cell lysates showed dose- and time-dependent patterns after LPS challenge.
- These findings suggest that changes in miRNA expression and their targets after LPS exposure could disturb normal uterine function through interference with cytokine and prostaglandin secretion
Conclusion or Significance
- This research illustrates the potential role of miRNAs in the body’s response to infection, in this case, LPS, a component of bacteria.
- It is suggested that these miRNAs might play a part in altering normal uterine functioning, hence contributing to reproductive issues in mares.
Cite This Article
APA
Ibrahim S, Szóstek-Mioduchowska A, Skarzynski D.
(2019).
Expression profiling of selected miRNAs in equine endometrium in response to LPS challenge in vitro: A new understanding of the inflammatory immune response.
Vet Immunol Immunopathol, 209, 37-44.
https://doi.org/10.1016/j.vetimm.2019.02.006 Publication
Researcher Affiliations
- Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research of PAS, Olsztyn, Poland; Department of Animal Reproduction and A.I, Veterinary Research Division, National Research Centre, Dokki, Giza, Egypt. Electronic address: sally_rashad2004@yahoo.com.
- Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research of PAS, Olsztyn, Poland. Electronic address: a.szostek-mioduchowska@pan.olsztyn.pl.
- Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research of PAS, Olsztyn, Poland. Electronic address: d.skarzynski@pan.olsztyn.pl.
MeSH Terms
- Animals
- Cell Survival
- Endometritis / genetics
- Endometritis / immunology
- Endometritis / veterinary
- Endometrium / cytology
- Endometrium / immunology
- Epithelial Cells / metabolism
- Female
- Gene Expression Profiling
- Gene Expression Regulation
- Horse Diseases / genetics
- Horse Diseases / immunology
- Horses
- Immunity, Innate
- Inflammation / genetics
- Inflammation / immunology
- Inflammation Mediators / metabolism
- Lipopolysaccharides
- MicroRNAs / biosynthesis
- MicroRNAs / genetics
- MicroRNAs / physiology
- Stromal Cells / metabolism
- Toll-Like Receptor 4 / genetics
Citations
This article has been cited 9 times.- Shokri A, Asadpour R, Jafari-Joozani R, Babaei E, Hajibemani A, Hamidian G. Plasma microRNAs as non-invasive biomarkers in bovine endometritis caused by Gram-negative and Gram-positive bacteria.. Vet Res Forum 2023;14(8):437-445.
- Wang B, Li L, Yu R. Exosomes From Adipose-Derived Stem Cells Suppress the Progression of Chronic Endometritis.. Cell Transplant 2023 Jan-Dec;32:9636897231173736.
- Ibrahim S, Hedia M, Taqi MO, Derbala MK, Mahmoud KGM, Ahmed Y, Sosa AS, Saber YHA, Hasanain MH, Nawito MF, Seidel GE. Extracellular vesicles in low volume uterine lavage and serum: novel and promising biomarker for endometritis in Arabian mares.. BMC Vet Res 2022 Jan 18;18(1):42.
- Liang Y, Ming Q, Shen T, Jin Y, Zhao X, Luo R, Wang J, Lu J. CircRNA circFADS2 is Downregulated in Endometritis and its Overexpression Promotes miR-643 Maturation in Human Endometrial Epithelial Cells to Suppress Cell Apoptosis.. Reprod Sci 2021 Dec;28(12):3508-3514.
- Ibrahim S, Hedia M, Taqi MO, Derbala MK, Mahmoud KGM, Ahmed Y, Ismail S, El-Belely M. Alterations in the Expression Profile of Serum miR-155, miR-223, miR-17, miR-200a, miR-205, as well as Levels of Interleukin 6, and Prostaglandins during Endometritis in Arabian Mares.. Vet Sci 2021 Jun 4;8(6).
- Oladejo AO, Li Y, Wu X, Imam BH, Shen W, Ding XZ, Wang S, Yan Z. MicroRNAome: Potential and Veritable Immunomolecular Therapeutic and Diagnostic Baseline for Lingering Bovine Endometritis.. Front Vet Sci 2020;7:614054.
- Sharma A, Shandilya UK, Sullivan T, Naylor D, Canovas A, Mallard BA, Karrow NA. Identification of Ovine Serum miRNAs Following Bacterial Lipopolysaccharide Challenge.. Int J Mol Sci 2020 Oct 25;21(21).
- Moussa Agha D, Rouas R, Najar M, Bouhtit F, Naamane N, Fayyad-Kazan H, Bron D, Meuleman N, Lewalle P, Merimi M. Identification of Acute Myeloid Leukemia Bone Marrow Circulating MicroRNAs.. Int J Mol Sci 2020 Sep 25;21(19).
- Paukszto L, Mikolajczyk A, Jastrzebski JP, Majewska M, Dobrzyn K, Kiezun M, Smolinska N, Kaminski T. Transcriptome, Spliceosome and Editome Expression Patterns of the Porcine Endometrium in Response to a Single Subclinical Dose of Salmonella Enteritidis Lipopolysaccharide.. Int J Mol Sci 2020 Jun 13;21(12).
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