Persimmon Leaves (Diospyros kaki) Extract Enhances the Viability of Human Corneal Endothelial Cells by Improving Na+-K+-ATPase Activity.

Ramsha Afzal,Hyung Bin Hwang

doi:10.3390/ph15010072

Abstract

The Na+/K+-ATPase, present in the basolateral membrane of human corneal endothelial cells (HCECs), is known to play an important role for corneal transparency. Na+/K+-ATPase dysfunction is one of the major causes of corneal decompensation. The ethanol extract of Diospyros kaki (EEDK) has been reported to increase corneal cell viability. Thus, we treated HCECs with EEDK and studied its effects on HCECs survival and Na+/K+-ATPase against cytotoxic drugs like staurosporine (ST) and ouabain (OU). Firstly, survival assays, (MTT assay and live dead-imaging) showed that decreased HCECs viability by ST and OU was significantly recovered by EEDK co-treatment. Secondly, Na+/K+-ATPase activity assays revealed that EEDK enhanced Na+/K+-ATPase enzymatic activity (* p < 0.01) with/without ST and OU. Finally, Na+/K+-ATPase expression analysis (Western Blot and confocal microscopy) demonstrated that EEDK treatment with/without ST and OU facilitates Na+/K+-ATPase expression in HCECs. Taken together, our findings led us to the conclusion that EEDK might aid HCECs survival in vitro by increasing the activity and expression of Na+/K+-ATPase enzyme. Since Na+/K+-ATPase activity is important to maintain cellular function of HCECs, we suggest that EEDK can be a potential effective agent against corneal edema and related corneal disorders.

Highlights

The cornea is a transparent outer layer of the eye that covers the entire front of the eye and focuses the light allowing for clear vision
Our results indicated that co-treatment with cytotoxic drugs had aapositive effect the viability ofofHCECs co-treatment of ethanol extract of Diospyros kaki (EEDK) with cytotoxic drugs had aand positive effect on(Figure the viability of human corneal endothelial cells (HCECs) compared to the cells exposed to as compared to the cells exposed to ST (Figure 2C,D) and OU alone (Figure 2E,F)
We proved that EEDK treatment increases HCECs viability, enzyme activity and expression of the Na+ /K+ -ATPase in cultured HCECs with/without cytotoxic agents

Summary

Introduction

The cornea is a transparent outer layer of the eye that covers the entire front of the eye and focuses the light allowing for clear vision. It consists of several layers of cells like the epithelium, Bowman’s layer, stroma, Descemet’s membrane, and endothelial layer. The corneal stroma makes up 90% of cornea and its hydration plays a major role in maintaining corneal transparency. The posterior side of the corneal stroma is lined by Descemet’s membrane and the innermost monolayer membrane of cornea is endothelium. The corneal endothelial cells are composed of hexagonal, squamous cells lining the posterior cornea. Apoptosis or necrosis of human corneal endothelial cells (HCECs) may lead to corneal edema and decompensation [1]

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