Purchase this article with an account.
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.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
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.
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.
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.
These findings indicate that AZM has a potential to suppress inflammatory responses stimulated by zymosan through a TLR2 pathway in human corneal epithelial cells.
This PDF is available to Subscribers Only