Document Detail

In vitro evidence of nerve growth factor effects on human conjunctival epithelial cell differentiation and mucin gene expression.
MedLine Citation:
PMID:  19407015     Owner:  NLM     Status:  MEDLINE    
PURPOSE: Mucins released into the tear film are crucial to maintaining a healthy ocular surface. Alterations in goblet cell numbers and mucin secretion are observed in chronic ocular surface inflammatory diseases. Nerve growth factor (NGF) plays a crucial role in healing and inflammation of the ocular surface. The aim of this study was to evaluate in vitro the effect of NGF on conjunctival goblet cell differentiation and mucin production and secretion. METHODS: Human conjunctival epithelial cells were exposed to increasing NGF concentrations (1 to 250 ng/mL) and analyzed to quantify cell growth (MTT/Ki67/BrdU), goblet cell differentiation (PAS/MUC5AC confocal staining), and mucin mRNA expression (real-time PCR). Secreted and cellular MUC5AC were also analyzed by sandwich-ELISA and FACS, respectively. To confirm the biological effects of NGF, the same evaluations were performed on primary cultures, and changes in markers of stemness (p63) and commitment (14-3-3 sigma) were also investigated. RESULTS: In cell cultures, NGF induced a dose-dependent increase of goblet cell numbers, MUC5AC production, storage, and release. Additionally, in primary cultures, NGF induced an increase of abortive colonies and 14-3-3 sigma protein, and a decrease of p63 mRNA and protein, suggesting a differentiating effect of NGF on human conjunctival epithelium. CONCLUSIONS: These findings show that NGF might play a role in the complex mechanism leading to conjunctival epithelium differentiation and mucin secretion. In addition to the known roles of NGF in promoting ocular surface healing and sensitivity, its effects on conjunctival goblet cells support a rationale to investigate the therapeutic effectiveness of NGF in dry eye disease.
Alessandro Lambiase; Alessandra Micera; Graziella Pellegrini; Daniela Merlo; Paolo Rama; Michele De Luca; Sergio Bonini; Stefano Bonini
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-04-30
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  50     ISSN:  1552-5783     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2009 Oct 
Date Detail:
Created Date:  2009-09-24     Completed Date:  2009-10-02     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  4622-30     Citation Subset:  IM    
CIR Laboratory of Ophthalmology, University Campus Bio-Medico, Rome, Italy.
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MeSH Terms
Blotting, Western
Cell Count
Cell Differentiation / drug effects*
Cell Proliferation / drug effects
Cells, Cultured
Conjunctiva / cytology*,  metabolism
Dose-Response Relationship, Drug
Enzyme-Linked Immunosorbent Assay
Exonucleases / metabolism
Flow Cytometry
Gene Expression Regulation / drug effects*
Goblet Cells / cytology*,  metabolism
Membrane Proteins / genetics,  metabolism
Microscopy, Confocal
Mucin 5AC / genetics*
Neoplasm Proteins / metabolism
Nerve Growth Factor / pharmacology*
RNA, Messenger / metabolism
Receptor, Nerve Growth Factor / metabolism
Receptor, trkA / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Tumor Markers, Biological / metabolism
Reg. No./Substance:
0/CKAP4 protein, human; 0/MUC5AC protein, human; 0/Membrane Proteins; 0/Mucin 5AC; 0/Neoplasm Proteins; 0/RNA, Messenger; 0/Receptor, Nerve Growth Factor; 0/Tumor Markers, Biological; 9061-61-4/Nerve Growth Factor; EC, trkA; EC 3.1.-/Exonucleases; EC 3.1.-/SFN protein, human

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