Ocular inflammation and corneal permeability alteration by benzalkonium chloride in rats: a protective effect of a myosin light chain kinase inhibitor

Invest Ophthalmol Vis Sci. 2013 Apr 17;54(4):2705-10. doi: 10.1167/iovs.12-10193.

Abstract

Purpose: The aim of this study was to evaluate the interest of an ophthalmic eyedrop preparation containing a myosin light chain kinase (MLCK) inhibitor, ML-7, in the treatment of ocular surface. The local protective effect on the inflammation and the increase of corneal permeability induced by benzalkonium (BAK) was evaluated.

Methods: An ocular instillation of 10 lL BAK at a concentration of 0.1% in PBS was performed on rats. The eyes were rinsed with sterilized water, 10 minutes after BAK preceded by instillation at T -24, -12, and -0.5 hours of 10 lL ofML-7: 100 μg (10 μL) into a gel form vehicle. All animals were sacrificed 6 hours after BAK instillation. The eyes were isolated for study in a masked manner. The ocular surface inflammation was assessed by measuring the inflammatory cell infiltration by a histologic quantitative analysis and for total ocular myeloperoxidase (MPO) activity. The tight junction permeability was tested.

Results: Instillation of 0.1% BAK increased the inflammation of the eye. The quantitative analysis showed an increase in the number of eosinophil and neutrophil polynuclears, and MPO activity. Pretreatment with ML-7 reduced inflammation (P < 0.05). The vehicle alone produced no notable effects. BAK instillation also thickened the fluorescent corneal front on frozen sections, indicating an increase of tight junction permeability. Pretreatment with ML-7 suppressed BAK-induced alterations of paracellular permeability while the vehicle had no visible effects.

Conclusions: Our study indicates that the inhibition of corneal cytoskeleton contraction by an MLCK inhibitor prevents BAK-induced ocular inflammatory response, and that ML-7 may be a new and original preparation in the treatment of ocular surface pathologies.

MeSH terms

  • Animals
  • Azepines / therapeutic use*
  • Benzalkonium Compounds / toxicity*
  • Cornea / drug effects*
  • Cornea / metabolism
  • Dry Eye Syndromes / drug therapy
  • Enzyme Inhibitors / therapeutic use*
  • Eosinophils / metabolism
  • Keratitis / chemically induced
  • Keratitis / metabolism
  • Keratitis / prevention & control*
  • Male
  • Myosin-Light-Chain Kinase / antagonists & inhibitors*
  • Naphthalenes / therapeutic use*
  • Neutrophils / metabolism
  • Ophthalmic Solutions
  • Permeability
  • Peroxidase / metabolism
  • Preservatives, Pharmaceutical / toxicity*
  • Rats
  • Rats, Wistar
  • Tight Junctions / drug effects*
  • Tight Junctions / metabolism

Substances

  • Azepines
  • Benzalkonium Compounds
  • Enzyme Inhibitors
  • Naphthalenes
  • Ophthalmic Solutions
  • Preservatives, Pharmaceutical
  • ML 7
  • Peroxidase
  • Myosin-Light-Chain Kinase