Kim Jeong Hun, Kim Jin Hyoung, Yu Young Suk, Min Bon-Hong, Kim Kyu-Won
Department of Ophthalmology, Seoul National University College of Medicine & Seoul Artificial Eye Center, Clinical Research Institute, Seoul National University Hospital, 28 Yongon-dong, Chongno-gu, Seoul 110-744, Korea.
Br J Ophthalmol. 2007 Nov;91(11):1541-6. doi: 10.1136/bjo.2007.115220. Epub 2007 May 2.
To assess the role of clusterin in retinal vascular development and in free radical damage in vivo and in vitro.
The expression of clusterin, von Willebrand factor (vWF), flk-1, heat shock protein 27 (Hsp27) and heat shock protein 70 (Hsp70) was examined in the retinas of developing mice and oxygen-induced retinopathy (OIR) mice by immunofluorescence staining and western blot analysis. Hydrogen peroxide (H(2)O(2))-pretreated human retinal endothelial cells (HREC) and astrocytes were cultured in the presence or absence of exogenous clusterin, and then the cell viability was measured using the MTT assay and DAPI staining.
Clusterin was expressed mainly in the inner retina and co-localised with vWF, an endothelial cell marker. During the mouse developmental process, clusterin expression was decreased, which was similar to the expression of flk-1, vWF and Hsp27. Furthermore, in the OIR model, clusterin expression changed in a similar way to both vWF and Hsp27. Under hypoxic conditions, clusterin expression increased in HREC and astrocytes. In H(2)O(2)-pretreated HREC and astrocytes, clusterin protected against apoptotic cell death.
These results suggest that clusterin is associated with protection from apoptotic retinal cell death in retinal development and in free radical damage.
评估簇集素在体内外视网膜血管发育及自由基损伤中的作用。
通过免疫荧光染色和蛋白质印迹分析，检测发育中小鼠和氧诱导视网膜病变（OIR）小鼠视网膜中簇集素、血管性血友病因子（vWF）、flk-1、热休克蛋白27（Hsp27）和热休克蛋白70（Hsp70）的表达。用过氧化氢（H₂O₂）预处理人视网膜内皮细胞（HREC）和星形胶质细胞，在有或无外源性簇集素存在的情况下进行培养，然后使用MTT法和DAPI染色测量细胞活力。
簇集素主要在内层视网膜表达，并与内皮细胞标志物vWF共定位。在小鼠发育过程中，簇集素表达降低，这与flk-1、vWF和Hsp27的表达情况相似。此外，在OIR模型中，簇集素的表达变化与vWF和Hsp27相似。在缺氧条件下，HREC和星形胶质细胞中簇集素表达增加。在H₂O₂预处理的HREC和星形胶质细胞中，簇集素可保护细胞免受凋亡性细胞死亡。
这些结果表明，簇集素与视网膜发育及自由基损伤中保护视网膜细胞免受凋亡性死亡有关。
