Abstract
Purpose:
To evaluate the anti-inflammatory effects of Resolvin-D1 (RV-D1) and its mode of action on human corneal epithelial (HCE) cells in vitro.
Methods:
Cultured HCE cells were incubated for different periods and at several concentrations of RV-D1. Oleic acid (OA) and Dexamethasone (DM) served as negative and positive controls, respectively. Cells were stimulated with polyriboinosinic : polyribocytidylic acid (poly I:C). The protein contents and mRNA expression levels of Tumor Necrosis Factor-α (TNF-α), Interleukin (IL)-6, IL-1β and IL-8 were evaluated with multiplex fluorescent bead immunoassay (FBI) and with real time-PCR, respectively. The expression of Inhibitory Factor-κBα (I-κBα) was evaluated with real time-PCR.
Results:
The protein contents of the pro-inflammatory cytokines TNF-α, IL-6, IL-1β and IL-8 were significantly increased after stimulation with Poly I:C. After incubation of the cells with RV-D1 at concentration of 1µM, the protein content of TNF-α decreased to 20.76±9.3% (p < 0.05), IL-6 to 43.54±14.16% (p < 0.001), IL-1β to 46.73±15.93% (p > 0.05) and IL-8 to 51.15±13.01% (p < 0.05), compared with stimulation with poly I:C alone. Similar results were demonstrated for the mRNA levels of each of these cytokines. The anti-inflammatory effects of RV-D1 were comparable to those of DM. The effects of RV-D1 on Poly I:C stimulated HCE cells were dose dependant for the reduction in protein contents of TNF-α, IL-6, IL-1β and IL-8. The decrease in the mRNA expression of TNF-α, IL-6, IL-1β and IL-8 was shown to be related to a decrease in the expression of I-κBα.
Conclusions:
RV-D1 may serve as a potent anti-inflammatory agent in ocular surface inflammation, as evaluated in cultured HCE. The anti-inflammatory effects of RV-D1 are comparable to those of corticosteroids, and are mediated through NF-κB signal transduction.
Keywords: 480 cornea: basic science •
482 cornea: epithelium •
557 inflammation