Laabich Aicha, Vissvesvaran Ganesh P, Lieu Kuo L, Murata Kyoko, McGinn Tim E, Manmoto Corinne C, Sinclair John R, Karliga Ibrahim, Leung David W, Fawzi Ahmad, Kubota Ryo
Acucela Inc., Seattle, Washington 98103, USA.
Invest Ophthalmol Vis Sci. 2006 Jul;47(7):3156-63. doi: 10.1167/iovs.05-1621.
The present study was performed to investigate the effect of crocin on blue light- and white light-induced rod and cone death in primary retinal cell cultures.
Primary retinal cell cultures were prepared from primate and bovine retinas. Fifteen-day-old cultures were exposed to blue actinic light or to white fluorescent light for 24 hours. Cultures were treated by the addition of different concentrations of crocin for 24 hours before light exposure or for 8 hours after light exposure. Cultures kept in the dark were used as controls. Green nucleic acid stain assay was used to evaluate cell death. Rods and cones were immunolabeled with specific antibodies and counted. TUNEL labeling was used to detect fragmented DNA in fixed cells after light exposure.
Primary retinal cell cultures contained a mixture of retinal cells enriched in photoreceptors, bipolar cells, and Müller cells. Twenty-four-hour exposure to blue and white light induced death in 70% to 80% of the photoreceptors in bovine and primate retinal cell cultures. Crocin protected the photoreceptors against blue light- or white light-mediated damage in a concentration-dependent manner with an EC50 of approximately 30 microM. TUNEL assays confirmed that crocin protected photoreceptors from light damage.
These results show that blue and white light selectively induce rod and cone cell death in an in vitro model. Crocin protects retinal photoreceptors against light-induced cell death.
本研究旨在探讨西红花苷对原代视网膜细胞培养物中蓝光和白光诱导的视杆细胞和视锥细胞死亡的影响。
从灵长类动物和牛的视网膜制备原代视网膜细胞培养物。将15日龄的培养物暴露于蓝光或白光下24小时。在光照前24小时或光照后8小时,添加不同浓度的西红花苷处理培养物。置于黑暗中的培养物用作对照。采用绿色核酸染色法评估细胞死亡情况。用特异性抗体对视杆细胞和视锥细胞进行免疫标记并计数。采用TUNEL标记法检测光照后固定细胞中的DNA片段化情况。
原代视网膜细胞培养物包含富含光感受器、双极细胞和穆勒细胞的视网膜细胞混合物。在牛和灵长类动物视网膜细胞培养物中,24小时的蓝光和白光暴露诱导70%至80%的光感受器死亡。西红花苷以浓度依赖的方式保护光感受器免受蓝光或白光介导的损伤,半数有效浓度(EC50)约为30微摩尔。TUNEL分析证实西红花苷可保护光感受器免受光损伤。
这些结果表明,在体外模型中,蓝光和白光可选择性诱导视杆细胞和视锥细胞死亡。西红花苷可保护视网膜光感受器免受光诱导的细胞死亡。