The therapeutic effect of DA-6034 on ocular inflammation via suppression of MMP-9 and inflammatory cytokines and activation of the MAPK signaling pathway in an experimental dry eye model

Curr Eye Res. 2010 Feb;35(2):165-75. doi: 10.3109/02713680903453494.

Abstract

Purpose: To investigate the effect of DA-6034, 7-carboxymethyloxy-3',4',5-trimethoxy flavone, in experimentally-induced inflammatory dry eye in rabbit. In addition, to elucidate the mechanism of DA-6034, we evaluated the mitogen-activated protein kinase (MAPK) signaling pathway and transcriptional factor-kappa B (NF-kB) in corneal epithelial cells.

Methods: Rabbit lacrimal glands were injected with the T-cell mitogen concanavalin A (Con A). DA-6034 was then administered topically four times a day for six days starting 24 hr after Con A injection. Tear volume, tear function, MMP-9 and inflammatory cytokine levels in the lacrimal glands, and histological evaluation were subsequently assessed. In in vitro study, phosphorylated MAPKs (c-Jun NH2-terminal kinase (JNK) and p38 MAPK) and NF-kB were detected by enzyme-linked immunosorbent assay (ELISA) using human corneal epithelial cells.

Results: A single injection of Con A into the lacrimal glands induced a pronounced inflammatory response, caused elevated levels of MMP-9 and cytokines IL-8 and TGF-beta(1), and induced a decrease in tear volume and shortening of tear breakup time (TBUT). In this inflammation model of dry eye, DA-6034 clearly showed therapeutic efficacy by restoring tear function and inhibiting inflammatory responses after topical ocular application. Furthermore, DA-6034 attenuated the phosphorylation of JNK and p38 MAPK and inhibited NF-kB activation in a concentration-dependent manner in corneal epithelial cells.

Conclusions: These results suggest that DA-6034 has the therapeutic effect in rabbit lacrimal gland inflammation model of dry eye and might be a potential treatment option for acute dry eye syndrome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Administration, Topical
  • Animals
  • Blotting, Western
  • Cell Culture Techniques
  • Cytokines / metabolism*
  • Dacryocystitis / drug therapy*
  • Dacryocystitis / enzymology
  • Disease Models, Animal*
  • Dry Eye Syndromes / drug therapy*
  • Dry Eye Syndromes / enzymology
  • Enzyme Activation
  • Enzyme-Linked Immunosorbent Assay
  • Epithelium, Corneal / metabolism
  • Flavonoids / therapeutic use*
  • MAP Kinase Signaling System / drug effects*
  • Matrix Metalloproteinase 9 / metabolism*
  • Mitogen-Activated Protein Kinases / metabolism*
  • NF-kappa B / metabolism
  • Phosphorylation
  • Rabbits
  • Tears / chemistry

Substances

  • Cytokines
  • Flavonoids
  • NF-kappa B
  • Mitogen-Activated Protein Kinases
  • Matrix Metalloproteinase 9
  • recoflavone