| Summary: | The Na<sup>+</sup>/K<sup>+</sup>-ATPase, present in the basolateral membrane of human corneal endothelial cells (HCECs), is known to play an important role for corneal transparency. Na<sup>+</sup>/K<sup>+</sup>-ATPase dysfunction is one of the major causes of corneal decompensation. The ethanol extract of <i>Diospyros kaki</i> (EEDK) has been reported to increase corneal cell viability. Thus, we treated HCECs with EEDK and studied its effects on HCECs survival and Na<sup>+</sup>/K<sup>+</sup>-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<sup>+</sup>/K<sup>+</sup>-ATPase activity assays revealed that EEDK enhanced Na<sup>+</sup>/K<sup>+</sup>-ATPase enzymatic activity (* <i>p</i> < 0.01) with/without ST and OU. Finally, Na<sup>+</sup>/K<sup>+</sup>-ATPase expression analysis (Western Blot and confocal microscopy) demonstrated that EEDK treatment with/without ST and OU facilitates Na<sup>+</sup>/K<sup>+</sup>-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<sup>+</sup>/K<sup>+</sup>-ATPase enzyme. Since Na<sup>+</sup>/K<sup>+</sup>-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.
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