Korean J Ophthalmol.  2020 Feb;34(1):1-10. 10.3341/kjo.2019.0046.

Effect of Diquafosol on Hyperosmotic Stress-induced Tumor Necrosis Factor-α and Interleukin-6 Expression in Human Corneal Epithelial Cells

Affiliations
  • 1Myungmoon Bio, Hwaseong, Korea.
  • 2Department of Physiology, College of Korean Medicine Dongguk University, Gyeongju, Korea.
  • 3Binaree, Daegu, Korea.
  • 4Central Ophthalmic Clinic, Daegu, Korea. eyepark9@naver.com
  • 5Division of Biomedicinal & Cosmetics, College of Sciences & Technology, Mokwon University, Daejeon, Korea.
  • 6Developmental Biology Laboratory, Department of Biology, College of Natural Sciences, Kyungpook National University, Daegu, Korea. jcjung@knu.ac.kr

Abstract

PURPOSE
Diquafosol is a pharmaceutical drug used for dry eye treatment with a novel mechanism of action. It is a purinergic P2Y2 receptor agonist that promotes the secretion of tears and healing of corneal epithelial wounds. However, its inhibitory effect on hyperosmotic stress-induced inflammation in human corneal epithelial cells (HCECs) remains unclear.
METHODS
A hyperosmotic stress model was established by transferring HCECs from isosmotic (312 mOsm/kg to hyperosmotic medium (500 mOsm/kg). HCECs were incubated with 500 mOsm/kg hyperosmotic medium for 30 minutes, and then treated with diquafosol (0.6-6 mg/mL) for 4 or 24 hours. Cells were then harvested and analyzed by western blot, immunocytochemistry, and real-time polymerase chain reaction to evaluate the expression of interleukin-6, tumor necrosis factor-alpha, and the phosphorylation status of nuclear factor-kappa B.
RESULTS
Diquafosol significantly decreased the mRNA and protein expression of hyperosmotic stress-induced tumor necrosis factor-alpha and interleukin-6. These results were supported by immunofluorescence staining and quantitative real-time polymerase chain reaction analysis. Furthermore, diquafosol inhibits nuclear factor-kappa B activation by suppressing the phosphorylation and degradation of the inhibitor of кB.
CONCLUSIONS
This study shows that diquafosol inhibits nuclear factor-kappa B signaling and inflammatory factors induced by hyperosmotic stress in HCECs. This suggests that using diquafosol for the improvement of dry eye syndrome could be effective in the treatment of inflammation-related corneal and conjunctival diseases.

Keyword

Diquafosol; Human corneal epithelial cells; Inflammation; Interleukin-6; Tumor necrosis factor-alpha

MeSH Terms

Blotting, Western
Conjunctival Diseases
Dry Eye Syndromes
Epithelial Cells*
Fluorescent Antibody Technique
Humans*
Immunohistochemistry
Inflammation
Interleukin-6*
Necrosis*
Phosphorylation
Real-Time Polymerase Chain Reaction
RNA, Messenger
Tears
Tumor Necrosis Factor-alpha
Wounds and Injuries
Interleukin-6
RNA, Messenger
Tumor Necrosis Factor-alpha
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