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MedLine Citation:
PMID:  25183764     Owner:  NLM     Status:  Publisher    
Purpose: To evaluate the role of TLR4 in corneal epithelial wound healing. Methods: The expression of TLR4 during in vivo corneal epithelial wound healing was examined by immunostaining in mice. The expression and activation of TLR4 was studied in primary or telomerase-immortalized human corneal epithelial cells (HCEC). Scratch assay was performed to evaluate in vitro wound closure using live time-lapse microscopy. Transwell migration assay and Ki67 immunostaining were done to evaluate migration and proliferation, respectively. Lipopolysaccharide (LPS) was used to activate TLR4 whereas CLI-095 was used for its inhibition. The expression of inflammatory cytokines was determined by RT-PCR and ELISA. The activation of p42/44 and p38 was determined by immunoblotting. Results: In the murine model, TLR4 immunostaining was prominently noted in the epithelium 8 hours after wounding. There was a 4-fold increase in the expression of TLR4 6 hours after in vitro scratch wounding (P<0.001). Confocal microscopy confirmed the membrane localization of TLR4/MD2 complex. There was a significant increase in migration, proliferation and wound closure in HCEC treated with LPS (P<0.05) while there was significant decrease with TLR4 inhibition (P<0.05). Addition of LPS to wounded HCEC resulted in a significant increase in the expression of IL-6, TNF-α, CXCL8/IL8, and CCL5/RANTES both at the mRNA and protein level. Likewise, LPS increased the activation of p42/44 and p38 in wounded HCEC. Conclusions: These results suggest that epithelial wounding induces the expression of functional TLR4. TLR4 signaling appears to contribute to early corneal epithelial wound repair by enhancing migration and proliferation.
Medi Eslani; Asadolah Movahedan; Neda Afsharkhamseh; Herve Sroussi; Ali R Djalilian
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-9-2
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  -     ISSN:  1552-5783     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2014 Sep 
Date Detail:
Created Date:  2014-9-3     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2014 by Association for Research in Vision and Ophthalmology.
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