Protective effect of clusterin on oxidative stress-induced cell death of human corneal endothelial cells.
OBJECTIVE
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.
Shin
Young Joo
YJ
Myung Dong St. Mary's Eye Center, Seoul National University College of Engineering, Seoul, Korea.
Kim
Jeong Hun
JH
Seo
Jong Mo
JM
Lee
Sang Mok
SM
Hyon
Joon Young
JY
Yu
Young Suk
YS
Wee
Won Ryang
WR
D000328:Adult; D053148:Caspase 3 / Q000378:metabolism; D002452:Cell Count; D016923:Cell Death; D002478:Cells, Cultured; D051152:Clusterin / Q000378:metabolism*; D019610:Cytoprotection; D042783:Endothelial Cells / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology*; D004728:Endothelium, Corneal / Q000473:pathology*; D005452:Fluoresceins / Q000378:metabolism; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D008856:Microscopy, Fluorescence; D018384:Oxidative Stress; D011419:Propidium / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D013194:Staining and Labeling
D051152:Clusterin; D005452:Fluoresceins; D017382:Reactive Oxygen Species; C029569:diacetyldichlorofluorescein; D011419:Propidium; D053148:Caspase 3