• The Korean Journal of Physiology and Pharmacology
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• v.18 no.2
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• pp.89-94
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• 2014

DA-6034, a eupatilin derivative of flavonoid, has shown potent effects on the protection of gastric mucosa and induced the increases in fluid and glycoprotein secretion in human and rat corneal and conjunctival cells, suggesting that it might be considered as a drug for the treatment of dry eye. However, whether DA-6034 induces $Ca^{2+}$ signaling and its underlying mechanism in epithelial cells are not known. In the present study, we investigated the mechanism for actions of DA-6034 in $Ca^{2+}$ signaling pathways of the epithelial cells (conjunctival and corneal cells) from human donor eyes and mouse salivary gland epithelial cells. DA-6034 activated $Ca^{2+}$-activated $Cl^-$ channels (CaCCs) and increased intracellular calcium concentrations ($[Ca^{2+}]_i$) in primary cultured human conjunctival cells. DA-6034 also increased $[Ca^{2+}]_i$ in mouse salivary gland cells and human corneal epithelial cells. $[Ca^{2+}]_i$ increase of DA-6034 was dependent on the $Ca^{2+}$ entry from extracellular and $Ca^{2+}$ release from internal $Ca^{2+}$ stores. Interestingly, these effects of DA-6034 were related to ryanodine receptors (RyRs) but not phospholipase C/inositol 1,4,5-triphosphate ($IP_3$) pathway and lysosomal $Ca^{2+}$ stores. These results suggest that DA-6034 induces $Ca^{2+}$ signaling via extracellular $Ca^{2+}$ entry and RyRs-sensitive $Ca^{2+}$ release from internal $Ca^{2+}$ stores in epithelial cells.

• Korean Journal of Veterinary Research
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• v.56 no.3
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• pp.155-160
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• 2016

This study was conducted to evaluate three different mixed formulations of sodium hyaluronate (SH) and carboxymethyl cellulose (CMC) using a low-humidity air flow-induced rat dry eye model and determine the most suitable mixture. The total thickness of the cornea, corneal epithelial thickness, corneal stroma thickness, damaged corneal epithelium percentage region, thickness of the bulbar conjunctiva epithelium, number of goblet cells, goblet cell occupation percentage region, and damaged bulbar conjunctiva epithelium percentage region were measured by histomorphological evaluation. After 5 h exposure to drying airflow, the thickness of the cornea and conjunctiva was decreased with desquamation of the corneal and conjunctiva epithelium. However, these dry eye symptoms were markedly inhibited by treatment with the reference and test formulations. More favorable effects on decreased thickness were detected in response to the CMC than the SH. However, SH had a greater protective effect against corneal and conjunctiva epithelial damage. The application of a mixture of 0.1% SH and 0.2% CMC showed more favorable effects on the corneal and conjunctival damage and the stabilization of the ocular surface than SH or CMC alone.

• BMB Reports
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• v.46 no.2
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• pp.124-129
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• 2013

FK506 binding protein 12 (FK506BP) belongs to a family of immunophilins, and is involved in multiple biological processes. However, the function of FK506BP in corneal disease remains unclear. In this study, we examined the protective effects on dry eye disease in a Botulinum toxin A (BTX-A) induced mouse model, using a cell-permeable PEP-1-FK506BP protein. PEP-1-FK506BP efficiently transduced into human corneal epithelial cells in a time- and dose-dependent manner, and remained stable in the cells for 48 h. In addition, we demonstrated that topical application of PEP-1-FK506BP was transduced into mouse cornea and conjunctiva by immunohistochemistry. Furthermore, topical application of PEP-1-FK506BP to BTX-A-induced mouse model markedly inhibited expression levels of pro-inflammatory cytokines such as interleukin-$1{\beta}$ (IL-$1{\beta}$), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and macrophage inhibitory factor (MIF) in corneal and conjunctival epithelium. These results suggest PEP-1-FK506BP as a potential therapeutic agent for dry eye diseases.