Anti-inflammatory effects of alpha linolenic acid on human corneal epithelial cells

Invest Ophthalmol Vis Sci. 2012 Jul 3;53(8):4396-406. doi: 10.1167/iovs.12-9724.

Abstract

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 linoleic acid (LA). Oleic acid (OA) and dexamethasone (DM) served as negative and positive controls, respectively. Cells were stimulated with either polyinosinic:polycytidylic acid (poly I:C) or lipopolysaccharide (LPS) complex. The protein contents and mRNA expression levels of IL-6, IL-8, IL-1β, and 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Inflammatory Agents, Non-Steroidal / pharmacology*
  • Blotting, Western
  • Cell Survival
  • Cells, Cultured
  • Dexamethasone / pharmacology
  • Epithelium, Corneal / drug effects*
  • Epithelium, Corneal / metabolism
  • Fluorescent Antibody Technique
  • Humans
  • I-kappa B Proteins / genetics
  • Interleukin-1beta / genetics
  • Interleukin-6 / genetics
  • Interleukin-8 / genetics
  • Lipopolysaccharides / pharmacology
  • NF-KappaB Inhibitor alpha
  • Oleic Acid / pharmacology
  • Poly I-C / pharmacology
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction
  • Tissue Donors
  • Tumor Necrosis Factor-alpha / genetics
  • alpha-Linolenic Acid / pharmacology*

Substances

  • Anti-Inflammatory Agents, Non-Steroidal
  • I-kappa B Proteins
  • IL6 protein, human
  • Interleukin-1beta
  • Interleukin-6
  • Interleukin-8
  • Lipopolysaccharides
  • NFKBIA protein, human
  • RNA, Messenger
  • Tumor Necrosis Factor-alpha
  • alpha-Linolenic Acid
  • NF-KappaB Inhibitor alpha
  • Oleic Acid
  • Dexamethasone
  • Poly I-C