The effects of 3% diquafosol sodium eye drop application on meibomian gland and ocular surface alterations in the Cu, Zn-superoxide dismutase-1 (Sod1) knockout mice

Keisuke Ikeda, Cem Simsek, Takashi Kojima, Kazunari Higa, Motoko Kawashima, Murat Dogru, Takahiko Shimizu, Kazuo Tsubota, Jun Shimazaki

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Purpose: The purpose of the study is to investigate the effect of 3% diquafosol sodium eye drops on meibomian gland and ocular surface alterations in the superoxide dismutase-1 (Sod1−/−) mice in comparison to the wild-type mouse. Methods: Three percent diquafosol sodium eye drop was instilled to 20 eyes of 10 50-week-old male Sod1−/− mice and 22 eyes of 11 C57BL/6 strain 50-week-old wild-type (WT) male mice six times a day for 2 weeks. Aqueous tear secretion quantity was measured with phenol red-impregnated cotton threads without anesthesia. Tear film stability and corneal epithelial damage were assessed by fluorescein and lissamine green staining. We also performed oil red O (ORO) lipid staining to evaluate the lipid changes in the meibomian glands. Meibomian gland specimens underwent hematoxylin and eosin staining to examine histopathological changes and meibomian gland acinar unit density after sacrifice. Immunohistochemistry staining was performed using cytokeratin 4, cytokeratin 13, and transglutaminase-1 antibodies. Quantitative real-time polymerase chain reaction for cytokeratin 4, cytokeratin 13, and transglutaminase-1 mRNA expression was also performed. Results: The aqueous tear quantity, the mean tear film breakup time, and the number of lipid droplets significantly improved in the Sod1−/− mice with treatment. The mean meibomian acinar unit density did not change in the Sod1−/− mice and WT mice after treatment. Application of 3% diquafosol sodium eye drop significantly decreased the corneal fluorescein and lissamine green staining scores in the Sod1−/− mice after 2 weeks. We showed a notable increase in cytokeratin 4, cytokeratin 13 immunohistochemistry staining, and cytokeratin 4, cytokeratin 13 mRNA expressions with a marked decrease in immunohistochemistry staining and significant decline in mRNA expression of transglutaminase-1 after 3% diquafosol sodium treatment. Conclusion: Topical application of 3% diquafosol sodium eye drop improved the number of lipid droplets, tear stability, and tear production which in turn appeared to have a favorable effect on the ocular surface epithelium. Three percent diquafosol sodium eye drop may be a potential treatment for age-related meibomian gland and dry eye disease based on the observations of the current study.

Original languageEnglish
Pages (from-to)1-12
Number of pages12
JournalGraefe's Archive for Clinical and Experimental Ophthalmology
Publication statusAccepted/In press - 2018 Feb 27



  • Diquafosol
  • Dry eye
  • Meibomian gland alterations
  • Oxidative stress

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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