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蓝光发光二极管诱导的小鼠视网膜变性的功能和形态学评估

Functional and morphological evaluation of blue light-emitting diode-induced retinal degeneration in mice.

作者信息

Kim Gyu Hyun, Kim Hyung Il, Paik Sun-Sook, Jung Sung Won, Kang Seungbum, Kim In-Beom

机构信息

Department of Anatomy, Catholic Neuroscience Institute, College of Medicine, The Catholic University of Korea, 505 Banpo-dong, Seocho-gu, Seoul, 137-701, Korea.

Gyeongju St. Mary's Eye Clinic, 293 Wonhwa-ro, Gyeongju-si, Gyeongsangbuk-do, 780-946, Korea.

出版信息

Graefes Arch Clin Exp Ophthalmol. 2016 Apr;254(4):705-16. doi: 10.1007/s00417-015-3258-x. Epub 2016 Jan 8.

Abstract

PURPOSE

The purpose of this study was to evaluate a retinal degeneration (RD) model induced by exposing mice to a blue light-emitting diode (LED), which led to photoreceptor cell death.

METHODS

RD was induced in BALB/c mice by exposure to a blue LED (460 nm) for 2 hours. Retinal function was examined using scotopic electroretinography (ERG). Histopathological changes were assessed by hematoxylin and eosin (H&E) staining and electron microscopy. Apoptotic cell death was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. In addition, retinal inflammation and oxidative stress were evaluated by immunohistochemistry with anti-glial fibrillary acidic protein (GFAP) and anti-8-hydroxy-2'-deoxyguanosine (8-OHdG), respectively.

RESULTS

Scotopic ERG showed that blue LED exposure resulted in a decrease in both a-waves and b-waves in mice retinas in an illuminance-dependent manner. H&E, TUNEL assay, and electron microscopy revealed massive photoreceptor cell death by apoptosis in the central region of the retina. Retinal stress and inflammation were detected by increased expression of GFAP and by electron microscopy findings demonstrating microglia infiltration in the outer nuclear layer and subretinal space. In addition, increased labeling of 8-OHdG was observed in the retinas from blue LED exposure.

CONCLUSIONS

These results suggest that blue LED-induced RD may be a useful animal model in which to study the pathogenesis of RD, including age-related macular degeneration, and to evaluate the effects of new therapeutic agents prior to clinical trials, where oxidative stress and inflammation are the underlying RD mechanisms.

摘要

目的

本研究旨在评估通过将小鼠暴露于蓝光发光二极管(LED)诱导的视网膜变性(RD)模型,该模型会导致光感受器细胞死亡。

方法

通过将BALB/c小鼠暴露于460纳米的蓝光LED下2小时来诱导RD。使用暗视视网膜电图(ERG)检查视网膜功能。通过苏木精和伊红(H&E)染色及电子显微镜评估组织病理学变化。通过末端脱氧核苷酸转移酶dUTP缺口末端标记(TUNEL)试验评估凋亡细胞死亡。此外,分别通过抗胶质纤维酸性蛋白(GFAP)免疫组织化学和抗8-羟基-2'-脱氧鸟苷(8-OHdG)免疫组织化学评估视网膜炎症和氧化应激。

结果

暗视ERG显示,蓝光LED暴露导致小鼠视网膜中a波和b波均以照度依赖的方式降低。H&E、TUNEL试验和电子显微镜显示视网膜中央区域大量光感受器细胞通过凋亡死亡。通过GFAP表达增加以及电子显微镜发现显示小胶质细胞浸润到外核层和视网膜下间隙,检测到视网膜应激和炎症。此外,在蓝光LED暴露的视网膜中观察到8-OHdG标记增加。

结论

这些结果表明,蓝光LED诱导的RD可能是一种有用的动物模型,可用于研究RD的发病机制,包括年龄相关性黄斑变性,并在临床试验前评估新治疗药物的效果,其中氧化应激和炎症是潜在的RD机制。

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