April 2009
Volume 50, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2009
Azithromycin Suppresses Pro-Inflammatory Mediators Stimulated by a TLR2 Ligand Zymosan in Human Corneal Epithelial Cells
Author Affiliations & Notes
  • N. Zhou
    Ocular Surface center, Cullen Eye Institute, Ophthalmology, Baylor College of Medicine, Houston, Texas
  • P. Ma
    Ocular Surface center, Cullen Eye Institute, Ophthalmology, Baylor College of Medicine, Houston, Texas
  • D.-Q. Li
    Ocular Surface center, Cullen Eye Institute, Ophthalmology, Baylor College of Medicine, Houston, Texas
  • S. C. Pflugfelder
    Ocular Surface center, Cullen Eye Institute, Ophthalmology, Baylor College of Medicine, Houston, Texas
  • Footnotes
    Commercial Relationships  N. Zhou, Inspire Pharmaceuticals, Inc, F; P. Ma, Inspire Pharmaceuticals, Inc, F; D.-Q. Li, Inspire Pharmaceuticals, Inc, F; S.C. Pflugfelder, Inspire Pharmaceuticals, Inc, F.
  • Footnotes
    Support  Inspire Pharmaceuticals, Inc., NIH grant EY11915 (SCP), DOD CDMRP PRMRP grant FY06 PR064719 (DQL), Research to Prevent Blindness, Oshman Foundation, William Stamps Farish Fund
Investigative Ophthalmology & Visual Science April 2009, Vol.50, 5545. doi:
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    • Get Citation

      N. Zhou, P. Ma, D.-Q. Li, S. C. Pflugfelder; Azithromycin Suppresses Pro-Inflammatory Mediators Stimulated by a TLR2 Ligand Zymosan in Human Corneal Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2009;50(13):5545.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : In addition to antibiotic effects, azithromycin (AZM) has been noted to have anti-inflammatory effects, particularly in the context of bacterial infections. The goal of this study was to explore the suppressive effects of AZM on pro-inflammatory mediators stimulated by toll like receptor (TLR) ligands, in human corneal epithelial cells.

Methods: : Primary human corneal epithelial cells were cultured from donor corneal limbal explants and grown to sub-confluence in SHEM. The cells were treated with extracted or synthetic microbial components, ligands for TLRs 2-6, respectively for 6-48 hours, with or without pre-incubation with azithromycin (1-50 µg/ml) or TLR antibodies. The cells were subjected to total RNA extraction, reverse transcription (RT) and real-time PCR using Taqman gene expression assays (Applied Biosystems). The medium supernatants of cells treated for 48 hours were collected for protein quantitation using Luminex immunobead assays.

Results: : The expression of pro-inflammatory cytokines (TNF- and IL-1β), chemokines (IL-8, RANTES), and matrix metalloproteinases (MMP-1 and -9) by human corneal epithelial cells was dramatically stimulated by zymosan, poly I:C (dsRNA) and flagellin, ligands for TLRs 2, 3 and 5 respectively, with peak stimulation at 16 hours. AZM, added 1 hour pre-stimulation, suppressed expression of these pro-inflammatory mediators stimulated by zymosan, but not by flagellin or poly I:C. TLR2 antibody or AZM each partially blocked the expression of TNF-, IL-1β, IL-8, RANTES, MMP-1 and MMP-9 induced by zymosan in human corneal epithelial cells.

Conclusions: : These findings indicate that AZM has a potential to suppress inflammatory responses stimulated by zymosan through a TLR2 pathway in human corneal epithelial cells.

Keywords: cornea: epithelium • inflammation • antibiotics/antifungals/antiparasitics 
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