TLR2/4 signaling pathway mediates sperm-induced inflammation in bovine endometrial epithelial cells in vitro.
Animals
Cattle
Cells, Cultured
Coculture Techniques
Dose-Response Relationship, Drug
Endometritis
/ metabolism
Endometrium
/ cytology
Epithelial Cells
/ metabolism
Female
Immunity, Innate
Immunohistochemistry
Inflammation
/ metabolism
Insemination, Artificial
Lipopolysaccharides
MAP Kinase Kinase 4
/ metabolism
Male
Phosphorylation
Signal Transduction
Spermatozoa
/ metabolism
Toll-Like Receptor 2
/ agonists
Toll-Like Receptor 4
/ agonists
p38 Mitogen-Activated Protein Kinases
/ metabolism
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2019
2019
Historique:
received:
28
09
2018
accepted:
15
03
2019
entrez:
18
4
2019
pubmed:
18
4
2019
medline:
28
12
2019
Statut:
epublish
Résumé
We have recently shown that sperm attachment to bovine endometrial epithelial cells (BEECs) triggers uterine local innate immunity with induction of a pro-inflammatory response in vitro, however details of the mechanism remain unknown. Here, we investigated the involvement of Toll-like receptor 2/4 (TLR2/4) pathway in mediating sperm-BEECs inflammatory process. Immunohistochemistry of the uterine tissue revealed that TLR2 and TLR4 proteins were present in the luminal and glandular epithelia of bovine endometrium. Moreover, BEECs monolayers were treated with TLR2 agonist (Pam; 0, 10, 100, and 1000 ng/ml) or TLR4 agonist (LPS; 0, 0.1, 1, and 10 ng/ml) for 0, 1, 3, or 6 h, followed by evaluating mRNA expression of the pro-inflammatory genes (TNFA, IL-1B, IL-8, and PGES) in BEECs using a real-time PCR. Both Pam and LPS treatments showed a dose-dependent stimulation of mRNA expression of the pro-inflammatory genes. To elucidate the functional role of TLR2/4 in sperm-BEECs interaction, BEECs monolayers were incubated with either TLR2 antagonist or TLR4 antibody for 2 h prior to the co-culture with sperm for 3 h. Importantly, pre-incubation of BEECs with TLR2 antagonist or TLR4 antibody prevented the stimulatory effect of sperm on the transcription of pro-inflammatory genes in BEECs. Furthermore, sperm increased the phosphorylation levels of TLR2/4 downstream targets (p38MAPK and JNK) in BEECs within 1 h of the co-culture. Treatment of BEECs with TLR2 antagonist prior to sperm addition inhibited JNK phosphorylation, while TLR4 antibody inhibited the phosphorylation of both p38MAPK and JNK. In conclusion, the present in vitro findings strongly suggest that bovine endometrial epithelial cells respond to sperm via TLR2/4 signal transduction.
Identifiants
pubmed: 30995239
doi: 10.1371/journal.pone.0214516
pii: PONE-D-18-28314
pmc: PMC6469758
doi:
Substances chimiques
Lipopolysaccharides
0
Toll-Like Receptor 2
0
Toll-Like Receptor 4
0
p38 Mitogen-Activated Protein Kinases
EC 2.7.11.24
MAP Kinase Kinase 4
EC 2.7.12.2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0214516Déclaration de conflit d'intérêts
The authors declare that there is no conflict of interest that could be preserved as prejudicing the impartiality of the research reported.
Références
Theriogenology. 1994 Nov 1;42(6):941-50
pubmed: 16727599
Nat Immunol. 2001 Aug;2(8):675-80
pubmed: 11477402
J Reprod Immunol. 2002 Oct-Nov;57(1-2):61-79
pubmed: 12385834
Biol Reprod. 2012 Feb 29;86(2):51
pubmed: 22053092
Rev Reprod. 2000 Sep;5(3):164-74
pubmed: 11006166
FASEB J. 2014 May;28(5):2358-68
pubmed: 24497579
J Clin Invest. 2005 Aug;115(8):2223-33
pubmed: 16025156
Anal Biochem. 1987 Apr;162(1):156-9
pubmed: 2440339
Immunol Lett. 2003 Jan 22;85(2):85-95
pubmed: 12527213
Endocrinology. 2014 Apr;155(4):1453-65
pubmed: 24437488
Reprod Domest Anim. 2008 Nov;43 Suppl 5:2-11
pubmed: 19068027
Proc Jpn Acad Ser B Phys Biol Sci. 2010;86(5):322-37
pubmed: 20467218
Immunobiology. 2008;213(3-4):205-24
pubmed: 18406368
J Biol Chem. 2007 Apr 13;282(15):11030-7
pubmed: 17308304
Clin Microbiol Rev. 2009 Apr;22(2):240-73, Table of Contents
pubmed: 19366914
Trends Immunol. 2003 Oct;24(10):534-9
pubmed: 14552837
Nat Rev Immunol. 2007 May;7(5):353-64
pubmed: 17457343
Biol Reprod. 2009 Oct;81(4):795-805
pubmed: 19494250
J Biol Chem. 2004 Feb 27;279(9):7370-7
pubmed: 14660645
Int Immunopharmacol. 2007 Aug;7(8):1111-21
pubmed: 17570328
J Exp Med. 2002 Jan 7;195(1):99-111
pubmed: 11781369
Nat Immunol. 2010 May;11(5):373-84
pubmed: 20404851
Annu Rev Immunol. 2002;20:197-216
pubmed: 11861602
Biol Reprod. 1988 Jun;38(5):1171-80
pubmed: 3408784
Reprod Sci. 2008 Apr;15(3):231-42
pubmed: 18421019
Angew Chem Int Ed Engl. 2012 Dec 3;51(49):12246-9
pubmed: 22969053
J Endocrinol. 2008 Dec;199(3):425-34
pubmed: 18824521
Theriogenology. 1998 Feb;49(3):607-18
pubmed: 10732039
Nature. 2010 Jun 17;465(7300):885-90
pubmed: 20485341
J Reprod Dev. 2013 Oct;59(5):470-8
pubmed: 23800958
Reprod Biol Endocrinol. 2008 Nov 18;6:53
pubmed: 19017375
Reprod Domest Anim. 2012 Aug;47 Suppl 5:31-41
pubmed: 22913558
Biol Reprod. 2009 Dec;81(6):1025-32
pubmed: 19439727
PLoS One. 2015 Jul 06;10(7):e0132336
pubmed: 26147469
Mol Reprod Dev. 2018 Mar;85(3):215-226
pubmed: 29337420
Methods. 2001 Dec;25(4):402-8
pubmed: 11846609
J Assist Reprod Genet. 2008 Jun;25(6):223-8
pubmed: 18509754
Development. 2008 Jun;135(11):2001-11
pubmed: 18434414
Biol Reprod. 2000 May;62(5):1116-20
pubmed: 10775156
Mol Hum Reprod. 2010 Mar;16(3):135-52
pubmed: 19880575
PLoS One. 2016 Sep 23;11(9):e0162309
pubmed: 27662642
J Biol Chem. 2002 Apr 26;277(17):15028-34
pubmed: 11836257