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Yoshiyuki Satake, Kazunari Higa, Murat Dogru, Hiroto Mitamura, Naoki Toriyama, Jun Shimazaki; The Effect of Rebamipide to the Human Conjunctival Epithelium. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3697.
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© ARVO (1962-2015); The Authors (2016-present)
Rebamipide has been reported to enhance mucosal protection in gastric ulcers. Recently, rebamipide eyedrops (Mucosta® Otsuka Pharmaceutical Co., Ltd, Tokyo) have become commercially available in Japan. The ocular effects of rebamipide are still unclear. We investigate the effect of rebamipide solution on cultivated human conjunctival cells.
Human limbal conjunctival tissues from donor cornea were cultured for two weeks to prepare the stratified conjunctival epithelial sheets. Rebamipide (5μM and 20μM) was added to the culture medium for the last four days of cultivation. Histlogical (H.E. and PAS staining) and immunohistochemiccal (K4, K13, MUC16, ZO-1, occludin, Ki67 and p63) analysis were performed. LC-biotin, as a tracer molecule, was also used in cultivated human conjunctival epithelial sheet to detect tight junction barrier function. The cell-cycle phase distributions of cultivated conjunctival cells were examined by flow cytometric analysis.
The addition of rebamipide (20μM) to the cultivation medium induced a more stratified cultivated human conjunctival epithelial sheet. With increasing stratification, ZO-1 and occludin were confirmed to be located in the surface layer of cultivated epithelial sheets, and the barrier function against the permeability of LC-biotin were also enhanced. The expressions of the proliferation markers including Ki67 and p63 were enhanced, and the cell fraction of G2 and M phases was also increased in a dose-dependent manner by the addition of rebamipide into the cultivation medium. There were no differences in PAS, K3 and K14 stainings in this cultivation system.
This study revealed acceleration of cultivated human conjunctival epithelial sheet stratification due to the increased cell proliferation by rebamipide, which enhanced the barrier function of the conjunctival epithelium.
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