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Keiko Yamada, Mayumi Ueta, Hiromi Nishigaki, Norihiko Yokoi, Chie Sotozono, Shigeru Kinoshita; The effects of rebamipide-based gene-expression analysis in human conjunctival epithelial cells. Invest. Ophthalmol. Vis. Sci. 2018;59(9):116. doi: https://doi.org/.
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It is reported that rebamipide (RBM), an eye-drop medicine used to treat dry eye in Japan, suppresses ocular-surface inflammation. The purpose of this present study was to determine the effects of RBM-based gene-expression analysis in primary human conjunctival epithelial cells (PHCjECs).
This study involved 3 female conjunctivochalasis (CCh) patients (mean age: 69±15 years) in whom informed consent was obtained after receiving a detailed explanation of the study protocols. PHCjECs were harvested from conjunctival tissues obtained and discarded at the time of CCh surgery. The harvested PHCjECs were cultured using purified dispase, were divided into two groups [i.e., polyI:C(10µg/ml) added group and polyI:C(10µg/ml)+RBM(2mM) added group], and were stimulated for 3 hours. An exhaustive gene-expression analysis was then performed to compare gene-expressions in both groups using the GeneChip™ Microarray System (Thermo Fisher Scientific, Waltham, MA).
GeneChip™ Microarray analysis showed that in the PHCjECs, eighteen transcripts [e.g., NmrA-like family domain containing 1 pseudogene (LOC344887), PR domain containing 1 with ZNF domain (PRDM1), and growth differentiation factor 15 (GDF 15)] were upregulated 1.5-fold more upon polyI:C stimulation with RBM than upon polyI:C stimulation without RBM in all 3 cases. In addition, six transcripts [e.g., myxovirus resistance 2 (MX2), RGM domain family-member B (RGMB), and v-myc myelocytomatosis viral oncogene homolog (nMyc)] were downregulated 0.6-fold less upon polyI:C stimulation with RBM than upon polyI:C stimulation without RBM in all 3 cases.
The findings of this study show that RBM was effective in both upregulating and suppressing various kinds of gene expressions in PHCjECs stimulated with polyI:C. Further study using quantitative real-time polymerase chain reaction is needed to confirm these upregulated or downregulated transcripts of their gene-expressions, and it is essential to elucidate the method by which each gene acts in the action mechanism of RBM on the ocular surface.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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