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Purpose:
Systemic polyunsaturated fatty acids (PUFAs) were shown to improve the symptoms of dry eye syndrome due to their anti-inflammatory effects. This study evaluated the in-vitro anti-inflammatory effects of PUFAs on human corneal epithelial (HCE) cells.
Methods:
HCE cells were incubated for 2 hours with different concentrations of PUFAs: Alpha-linolenic acid (ALA), Gamma-linolenic acid (GLA) and Linolenic acid (LA). Oleic acid (OA) and dexamethasone (DM) served as negative and positive controls, respectively. Cells were stimulated with either polyinosinic
olycytidylic acid (poly I:C) or lipopolysaccharide (LPS) complex. The protein contents and mRNA expression levels of Interleukin (IL)-6, IL-8, IL-1β, and Tumor necrosis factor-α (TNF-α) were evaluated with multiplex fluorescent bead immunoassay and real time-PCR, respectively. The expression of inhibitory factor-κBα (I-κBα) was evaluated with real time-PCR.
Results:
The protein and mRNA levels of IL-6, IL-8, IL-1β and TNF-α were significantly increased after stimulation with LPS or poly I:C. Following treatment with ALA, a significant decrease was demonstrated in the protein content of TNF-α to 23.81% (P<0.001), IL-6 to 46.71% (P<0.001), IL-1β to 20.86% (P<0.05) and IL-8 to 52.21% (P<0.001). Similar results were demonstrated at the mRNA level. The anti-inflammatory effects of ALA were similar to those of DM for all of the pro-inflammatory cytokines. The ALA inhibition of the pro-inflammatory cytokines was associated with a significant reduction of I-κBα.
Conclusions:
ALA may serve as a potent anti-inflammatory agent in ocular surface inflammation. The anti-inflammatory effects of ALA are comparable to those of corticosteroids, and are mediated through NF-κB signal transduction.
Systemic polyunsaturated fatty acids (PUFAs) were shown to improve the symptoms of dry eye syndrome due to their anti-inflammatory effects. This study evaluated the in-vitro anti-inflammatory effects of PUFAs on human corneal epithelial (HCE) cells.
Methods:
HCE cells were incubated for 2 hours with different concentrations of PUFAs: Alpha-linolenic acid (ALA), Gamma-linolenic acid (GLA) and Linolenic acid (LA). Oleic acid (OA) and dexamethasone (DM) served as negative and positive controls, respectively. Cells were stimulated with either polyinosinic
olycytidylic acid (poly I:C) or lipopolysaccharide (LPS) complex. The protein contents and mRNA expression levels of Interleukin (IL)-6, IL-8, IL-1β, and Tumor necrosis factor-α (TNF-α) were evaluated with multiplex fluorescent bead immunoassay and real time-PCR, respectively. The expression of inhibitory factor-κBα (I-κBα) was evaluated with real time-PCR.Results:
The protein and mRNA levels of IL-6, IL-8, IL-1β and TNF-α were significantly increased after stimulation with LPS or poly I:C. Following treatment with ALA, a significant decrease was demonstrated in the protein content of TNF-α to 23.81% (P<0.001), IL-6 to 46.71% (P<0.001), IL-1β to 20.86% (P<0.05) and IL-8 to 52.21% (P<0.001). Similar results were demonstrated at the mRNA level. The anti-inflammatory effects of ALA were similar to those of DM for all of the pro-inflammatory cytokines. The ALA inhibition of the pro-inflammatory cytokines was associated with a significant reduction of I-κBα.
Conclusions:
ALA may serve as a potent anti-inflammatory agent in ocular surface inflammation. The anti-inflammatory effects of ALA are comparable to those of corticosteroids, and are mediated through NF-κB signal transduction.
Erdinest N, Shmueli O, Grossman Y, Ovadia H, Solomon A.
Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.