Protective effect of clusterin on oxidative stress-induced cell death of human corneal endothelial cells

PURPOSE: To investigate the protective effect of clusterin on oxidative stress-induced cell death of human corneal endothelial cells. METHODS: Human corneal endothelial cells (HCECs) were cultured according to previously published methods. With treatment of various concentrations (0-50 mM) of tert-butyl hydroperoxide (tBHP) or clusterin, reactive oxygen species (ROS) producrion was measured using an oxidationsensitive fluorescent probe and 2′7′-dichlorofluorescin diacetate (DCFH-DA). Cell viability was assayed with a Cell Counting Kit-8. RESULTS: In HCECs, DCF-DA staining revealed that cells treated with a higher concentration of tBHP had higher fluorescent intensity than cells treated with clusterin, compared to control cells. Clusterin significantly inhibited tBHP-induced ROS production. Cell viability decreased with higher tBHP concentration. Cells treated with clusterin had higher viability than control cells at 5 mM tBHP. Clusterin effectively protected HCECs from ROS-induced cell death. CONCLUSIONS: Our data suggest that clusterin may protect HCECs from oxidative injury-mediated cell death via inhibition of ROS production.