摘要
目的观察蓝光对人视网膜色素上皮细胞凋亡的影响并探讨其可能机制。方法细胞株分为蓝光照射组和对照组。蓝光照射组,利用35 W白色冷光灯加用蓝色滤光片建立蓝光损伤体外培养的RPE细胞模型,蓝光控制波长范围470~520 nm,光照强度在2000 lx左右,光照时间24~96 h,实验过程中保证光照在细胞培养孵箱内密进行。对照组为常规避光孵箱培养细胞,除无光线照射外,各培养条件相同。利用流式细胞仪检测RPE细胞的凋亡变化,利用流式细胞仪和激光共聚焦显微镜检测RPE细胞内游离钙离子浓度。结果与对照组比较,蓝光照射组的RPE细胞凋亡增加(P<0.05);照射组的RPE细胞内游离钙离子浓度增加(P<0.05);通过特异性抑制L型电压依赖性钙离子通道功能降低RPE细胞内游离钙离子浓度,在一定程度上降低了蓝光引起的RPE细胞凋亡。结论蓝光可以通过增加RPE细胞内游离钙离子浓度促进RPE细胞的凋亡,L型电压依赖性钙离子通道有望成为治疗年龄相关性黄斑变性等视网膜变性疾病的新靶点。
Objective To investigate the influence and mechanism of blue light on the apoptosis of human retinal pigment epithelial cells. Methods Cells were divided into blue light group and control group. 35 W white light lamp with blue filter was used to establish damaged RPE cell model in vitro, blue ray wave length ranged 470-520 nm, and the light intensity was about 2000 lx, light exposure time was set to 24-96 h, in the experimental process, the light was carried out in the cell culture incubation box. The control group cells were cultured in the conventional light avoidance incubator, and the culture conditions were as the same as the blue light group except without light. After exposure to blue light, the apoptosis of human retinal pigment epithelial cells was tested by flow cytometry. And then the intracellular calcium concentration of human retinal pigment epithelial cells was measured by flow cytometry and laser scanning confocal microscope. Results Compared with control group, the apoptosis of human retinal pigment epithelial cells increased in blue light group(P < 0.05), and the intracellular calcium concentration of human retinal pigment epithelial cells increased(P < 0.05). The intracellular calcium concentration of human RPE cells decreased by specific inhibition of L-type voltage-dependent calcium channel, RPE cell apoptosis caused by blue light decreased. Conclusion Blue light promotes apoptosis of human retinal pigment epithelial cells by the up-regulation of intracellular calcium concentration.
引文
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