簇集素对氧化应激诱导的人角膜内皮细胞死亡的保护作用。
Protective effect of clusterin on oxidative stress-induced cell death of human corneal endothelial cells.
作者信息
Shin Young Joo, Kim Jeong Hun, Seo Jong Mo, Lee Sang Mok, Hyon Joon Young, Yu Young Suk, Wee Won Ryang
机构信息
Myung Dong St. Mary's Eye Center, Seoul National University College of Engineering, Seoul, Korea.
出版信息
Mol Vis. 2009 Dec 16;15:2789-95.
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.
目的
研究簇集素对氧化应激诱导的人角膜内皮细胞死亡的保护作用。
方法
按照先前发表的方法培养人角膜内皮细胞（HCECs）。用不同浓度（0 - 50 mM）的叔丁基过氧化氢（tBHP）或簇集素处理后，使用氧化敏感荧光探针和2'7'-二氯荧光素二乙酸酯（DCFH-DA）测量活性氧（ROS）的产生。用细胞计数试剂盒-8检测细胞活力。
结果
在HCECs中，DCF-DA染色显示，与对照细胞相比，用较高浓度tBHP处理的细胞比用簇集素处理的细胞具有更高的荧光强度。簇集素显著抑制tBHP诱导的ROS产生。细胞活力随tBHP浓度升高而降低。在5 mM tBHP时，用簇集素处理的细胞比对照细胞具有更高的活力。簇集素有效地保护HCECs免受ROS诱导的细胞死亡。
结论
我们的数据表明，簇集素可能通过抑制ROS产生来保护HCECs免受氧化损伤介导的细胞死亡。
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