1 Department of Ophthalmology, Okayama University Medical School and Graduate School of Medicine , Dentistry, and Pharmaceutical Sciences, Okayama City, Japan .
2 Department of Medical Neurobiology, Okayama University Medical School and Graduate School of Medicine , Dentistry, and Pharmaceutical Sciences, Okayama City, Japan .
J Ocul Pharmacol Ther. 2017 Apr;33(3):149-160. doi: 10.1089/jop.2016.0093. Epub 2017 Jan 13.
Our previous study demonstrated that photoelectric dye-coupled polyethylene film (Okayama University-type retinal prosthesis), which was implanted in subretinal space of the eyes of Royal College of Surgeons (RCS) rats, prevented retinal neurons from apoptotic death. In this study, we aimed to examine whether photoelectric dye itself would protect retinal neurons from apoptosis in RCS rats.
RCS rats received intravitreous injection of different concentrations of the dye in the left eye and housed under a 12-h light-dark cycle. Saline injection in the right eye served as control. In addition, RCS rats with dye injection were kept in 24-h daily dark condition. Sections were processed for terminal deoxynucleotidyl transferase-mediated fluorescein-conjugated-dUTP nick-end-labeling (TUNEL) assay and immunohistochemical staining of glial fibrillary acidic protein (GFAP) and protein kinase Cα (PKCα).
The number of TUNEL-positive cells significantly decreased in the retina of dye-injected eyes compared with those in saline-injected eyes (P = 0.0001, 2-factor analysis of variance [ANOVA]), under 12-h light-dark cycle. Significant decrease of TUNEL-positive cells was noted in the retina of rats with dye injection compared with those with saline injection, kept under 24-h dark condition (P = 0.0001, 2-factor ANOVA). Immunoreactive area for GFAP decreased significantly in the retina of dye-injected eyes compared with that in controls (P = 0.0001, 2-factor ANOVA), whereas immunoreactive area for PKCα increased significantly in the retina of dye-injected eyes compared with that in controls (P = 0.01, 2-factor ANOVA).
Photoelectric dye inhibits apoptotic death of photoreceptor cells in RCS rats and downregulates GFAP expression in retinal Müller cells. Photoelectric dye may be a candidate agent for neuroprotection in retinitis pigmentosa and other retinal diseases.
我们之前的研究表明,植入 Royal College of Surgeons(RCS)大鼠视网膜下腔的光电染料偶联聚乙烯膜(冈崎大学型视网膜假体)可防止视网膜神经元发生凋亡性死亡。在这项研究中,我们旨在研究光电染料本身是否会保护 RCS 大鼠的视网膜神经元免受凋亡。
RCS 大鼠左眼接受不同浓度染料的玻璃体腔内注射,并在 12 小时光照-黑暗循环下饲养。右眼注射生理盐水作为对照。此外,接受染料注射的 RCS 大鼠保持在 24 小时的每日黑暗环境中。对切片进行末端脱氧核苷酸转移酶介导的荧光素-dUTP 缺口末端标记(TUNEL)检测和胶质纤维酸性蛋白(GFAP)和蛋白激酶 Cα(PKCα)的免疫组织化学染色。
在 12 小时光照-黑暗循环下,与生理盐水注射眼相比,染料注射眼的视网膜中 TUNEL 阳性细胞的数量显著减少(P=0.0001,双因素方差分析[ANOVA])。在 24 小时黑暗条件下,与生理盐水注射组相比,染料注射组的视网膜 TUNEL 阳性细胞数量显著减少(P=0.0001,双因素 ANOVA)。与对照组相比,染料注射眼的视网膜中 GFAP 免疫反应性面积显著减少(P=0.0001,双因素 ANOVA),而染料注射眼的视网膜中 PKCα 免疫反应性面积显著增加(P=0.01,双因素 ANOVA)。
光电染料可抑制 RCS 大鼠光感受器细胞的凋亡性死亡,并下调视网膜 Müller 细胞中的 GFAP 表达。光电染料可能是治疗色素性视网膜炎和其他视网膜疾病的神经保护候选药物。