| Increased platelet-activating factor receptor gene expression by corneal epithelial wound healing. | |
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MedLine Citation:
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PMID: 10845588 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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PURPOSE: Platelet activating factor (PAF) is a potent inflammatory mediator the synthesis of which increases in the cornea after injury. The effects of PAF are mediated by receptors (PAF-R), which are present in target cells. This study was undertaken to investigate the effects of wound healing, PAF, and growth factors on modulating PAF-R mRNA levels in corneal epithelial cells. METHODS: Cultures of rabbit corneal epithelial (RCE), rabbit limbal epithelial (RLE), rabbit corneal fibroblast (RCF), and rabbit corneal endothelial (RCEn) cells, as well as rabbit corneal keratocytes (RCKs) were used. For the in vivo wound-healing experiments, a 7-mm central corneal deepithelialization was performed in anesthetized rabbits. For the in vitro experiments, wounded rabbit corneas were maintained in organ culture. Corneas were stimulated with 120 nM PAF or preincubated with PAF antagonists, cyclohexamide (CHX) or actinomycin D (AcD) before adding PAF. RCE cells were stimulated with transforming growth factor (TGF)-beta1, -beta2, and, -beta3, basic fibroblast growth factor (bFGF), keratinocyte growth factor (KGF); and hepatocyte growth factor (HGF). Total RNA was isolated and PAF-R expression evaluated by reverse transcription-polymerase chain reaction (RT-PCR), Northern blot analysis, and quantitative RT-PCR. RESULTS: PAF-R mRNA was expressed in RCE, RLE, and RCEn cells and RCKs, but not in RCFs. After epithelial injury, PAF-R expression increased from 2.5 to 4 times, both in vitro and in vivo. Addition of cPAF further stimulated PAF-R gene expression in epithelium, which was abolished by PAF antagonists. Quantitative RT-PCR revealed that PAF stimulated PAF-R mRNA threefold after injury. The induction of PAF-R by its agonist required previous injury and was inhibited by AcD but not by CHX. Treatment of RCE cells with TGF-beta1, -beta2, or -beta3, HGF, and KGF increased mRNA in PAF-R; however, bFGF had no effect. CONCLUSIONS: Corneal injury produces changes in PAF-R mRNA expression. Whereas stroma fibroblastic cells lost the PAF-R gene expression found in keratocytes, corneal epithelial injury upregulated PAF-R mRNA. These results suggest that activation of selective growth factors and increases in PAF synthesis after injury stimulate PAF-R gene transcription and constitute important feedback mechanisms needed to maintain the inflammatory process and regulate epithelial wound healing. |
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Authors:
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X Ma; H E Bazan |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Investigative ophthalmology & visual science Volume: 41 ISSN: 0146-0404 ISO Abbreviation: Invest. Ophthalmol. Vis. Sci. Publication Date: 2000 Jun |
Date Detail:
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Created Date: 2000-06-14 Completed Date: 2000-06-14 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 7703701 Medline TA: Invest Ophthalmol Vis Sci Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 1696-702 Citation Subset: IM |
Affiliation:
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Department of Ophthalmology and Neuroscience Center, Louisiana State University Health Sciences Center, New Orleans, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Blotting, Northern Cells, Cultured Cornea / drug effects, injuries Epithelium, Corneal / drug effects, metabolism* Fibroblast Growth Factor 10 Fibroblast Growth Factor 2 / pharmacology Fibroblast Growth Factor 7 Fibroblast Growth Factors* Fibroblasts / drug effects, metabolism Gene Expression* Growth Substances / pharmacology Hepatocyte Growth Factor / pharmacology Organ Culture Techniques Platelet Activating Factor / antagonists & inhibitors, pharmacology Platelet Membrane Glycoproteins / biosynthesis, genetics* RNA, Messenger / biosynthesis* Rabbits Receptors, Cell Surface* Receptors, G-Protein-Coupled* Reverse Transcriptase Polymerase Chain Reaction Transforming Growth Factor beta Up-Regulation Wound Healing* |
| Grant Support | |
ID/Acronym/Agency:
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04928//PHS HHS; EY02377/EY/NEI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Fibroblast Growth Factor 10; 0/Growth Substances; 0/Platelet Activating Factor; 0/Platelet Membrane Glycoproteins; 0/RNA, Messenger; 0/Receptors, Cell Surface; 0/Receptors, G-Protein-Coupled; 0/Transforming Growth Factor beta; 0/platelet activating factor receptor; 103107-01-3/Fibroblast Growth Factor 2; 126469-10-1/Fibroblast Growth Factor 7; 62031-54-3/Fibroblast Growth Factors; 67256-21-7/Hepatocyte Growth Factor |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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