Abstract
Intestinal subepithelial myofibroblasts (ISMFs) are mesenchymal cells that reside in the subepithelial region throughout the intestine. When the intestine is damaged, the migratory and mitotic responses of ISMFs are crucial for wound closure. However, their mechanism of action remains unknown. We have investigated the role of cyclooxygenase (COX) and its metabolite prostaglandin E2 (PGE2) in the wound repair process of bovine ISMFs. The action of a mechanical scratch in a layer of ISMFs in cell culture elevated the levels of both COX-2 mRNA expression and PGE2 secretion 1 and 6 h after the event. After 24 h ISMFs had migrated to and reduced the wounded area around the site of the scratch. Treatment with the COX-1/2 inhibitor indomethacin, the COX-2 inhibitor 3-(4-methylsulphonylphenyl)-4-phenyl-5-trifluoromethylisoxazole (CAY10404), or E prostanoid receptor 2 to 4 (EP2–EP4) antagonists significantly inhibited wound repair. Conversely, inhibition of wound closure by indomethicin was reversed by treatment with PGE2 or agonists of the receptors EP2, EP3, or EP4 but not of EP1. Although EP2 to EP4 stimulation did not influence ISMF proliferation, it did stimulate ISMF migration in the transwell cell migration assay. It is noteworthy that cell migration stimulated by EP2 and EP4 was inhibited by the tyrosine kinase receptor inhibitor genistein and also by (Z)-3-[2,4-dimethyl-5-(2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-1H-pyrrol-3-yl]-propionic acid (SU6668). However, cell migration stimulated by EP3 was unaffected. Reverse transcription-polymerase chain reaction showed EP2 or EP4 stimulation elevated the level of mRNA expression for fibroblast growth factor-2, which stimulates ISMF migration. Collectively, COX-2-dependent PGE2 secretion promotes wound healing by ISMFs. PGE2-EP3 signaling may directly stimulate ISMF migration. PGE2-EP2/4 signaling indirectly stimulates ISMF migration by elevating the level of growth factor secretion.
Footnotes
This work was supported by Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists and the Japan Society for the Promotion of Science.
Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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ABBREVIATIONS:
- ISMF
- intestinal subepithelial myofibroblast
- PG
- prostaglandin
- PGE2
- prostaglandin E2
- PGES
- PGE2 synthase
- cPGES
- cytosolic PGES
- mPGES
- microsomal PGES
- EP
- E prostanoid receptor
- COX
- cyclooxygenase
- FGF
- fibroblast growth factor
- GAPDH
- glyceraldehyde-3-phosphate dehydrogenase
- VEGF
- vascular endothelial growth factor
- HGF
- hepatocyte growth factor
- RT-PCR
- reverse transcription-polymerase chain reaction
- FBS
- fetal bovine serum
- DMEM
- Dulbecco's modified Eagle's medium
- bp
- base pairs
- CAY10404
- 3-(4-methylsulphonylphenyl)-4-phenyl-5-trifluoromethylisoxazole
- SU6668
- (Z)-3-[2,4-dimethyl-5-(2-oxo-1,2-dihydro-indol-3-ylidenemethyl)-1H-pyrrol-3-yl]-propionic acid
- AH6809
- 6-isopropoxy-9-oxoxanthene-2-carboxylic acid
- AH23848
- (7-[5α-([1S,1α(Z)-biphenyl]-4-ylmethoxy)-2β-(4-morpholinyl)-3-oxocyclopentyl]-4-heptenoic acid, calcium salt), hydrate
- MK886
- 1-[(4-chlorophenyl)methyl]-3-[(1,1-dimethylethyl)thio]-α,α-dimethyl-5-(1-methylethyl)-1H-indole-2-propanoic acid, sodium salt
- SC-560
- 5-(4-chlorophenyl)-1-(4-methoxyphenyl)-3-(trifluoromethyl)-1H-pyrazole
- LY798106
- N-[(5-bromo-2-methoxyphenyl)sulfonyl]-3-[2-(2-naphthalenylmethyl)phenyl]-2-propenamide
- ONO-DI-004
- 17S-2,5-ethano-6-oxo-17,20-dimethyl prostaglandin E1
- ONO-AE1-259-01
- 11,15-O-dimethyl prostaglandin E2
- ONO-AE-248
- 16S-9-deoxy-9β-chloro-15-deoxy-16-hydroxy-17,17-trimethylene-19,20-didehydro prostaglandin F2
- ONO-AE1-329
- 16-(3-methoxymethyl) phenyl-ω-tetranor-3,7-dithia-prostaglandin E1.
- Received November 8, 2011.
- Accepted December 1, 2011.
- Copyright © 2012 by The American Society for Pharmacology and Experimental Therapeutics
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