Protective effects of NSP-116, a novel imidazolyl aniline derivative, against light-induced retinal damage in vitro and in vivo
Protective effects of NSP-116, a novel imidazolyl aniline derivative, against light-induced retinal damage in vitro and in vivo
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DOI：
10.1016/j.freeradbiomed.2016.03.036
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发表时间：
2016-07-01
影响因子：
7.4
通讯作者：
Hara, Hideaki
中科院分区：
文献类型：
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作者：
Izawa, Hiroshi;Shimazawa, Masamitsu;Hara, Hideaki
在本研究中，我们研究了一种新型的咪唑基苯胺衍生物NSP116[4-(4-acetylpiperazin-1-yl)-2-(1H-imidazol-1-yl)苯胺对光诱导的光感受器细胞损伤的保护作用。在体外实验中，小鼠光感受器(661W)细胞受到光照24小时的损伤，用Hoechst 33342/碘化丙啶核染色法和四唑盐(WST-8)比色法检测光照后661W细胞的存活率。用活性氧(ROS)敏感探针5-(和6)-氯甲基-2，7-二氯二氢荧光素二乙酸酯(CM-H(2)DCFDA)检测661W细胞内自由基的产生。NSP-116显著抑制光诱导的661W细胞死亡和ROS的产生。在活体小鼠实验中，暗适应后暴露于8000lx的白光3h可引起视网膜损伤。在光照后第5天记录视网膜电信号并测量外核层(ONL)厚度，以评估视网膜损伤。光暴露前单次口服NSP-116可保护视网膜功能和光暴露后ONL变薄。此外，用硫代巴比妥酸反应物质(TBARS)法检测NSP-116对猪视网膜脂质过氧化的影响，发现NSP-116可减少TBARS的产生。电子自旋共振(ESR)测试表明，NSP-116对1，1-二苯基-2-苦基肼(DPPH)自由基、超氧阴离子自由基(中心点O-2(-))和羟基自由基(中心点OH)具有清除活性。这些结果表明，NSP-116作为一种自由基清除剂，在体内外对光诱导的光感受器变性具有保护作用，可能是一种治疗视网膜退行性疾病的新药物，如干性年龄相关性黄斑变性(AMD)。(C)2016 Elsevier Inc.保留所有权利。
In this study, we investigated the protective effects of NSP-116 [4-(4-acetylpiperazin-1-yl)-2-(1H-imidazol-1-yl) aniline], a novel imidazolyl aniline derivative, against light-induced photoreceptor cell damage. In an in vitro experiment, murine photoreceptor (661 W) cells were damaged by exposure to light for 24 h. Viability of 661 W cells after light exposure was assessed by Hoechst 33342/Propidium iodide nuclear staining and a tetrazolium salt (WST-8) assay. Intracellular radical production in 661 W cells was evaluated using the reactive oxygen species (ROS) sensitive probe 5-(and 6)-chloromethyl-2, 7-di-chlorodihydrofluorescein diacetate acetyl ester (CM-H(2)DCFDA). NSP-116 significantly suppressed light induced cell death and ROS production in 661 W cells. In an in vivo mouse experiment, retinal damage was induced by exposure to white light at 8000 lx for 3 h after dark adaptation. Retinal damage was evaluated by recording the electroretinogram and measuring the outer nuclear layer (ONL) thickness at 5 days after light exposure. Single oral administration of NSP-116 before light exposure protected retinal function and ONL thinning after light exposure. Furthermore, the effect of NSP-116 on lipid peroxidation was evaluated using thiobarbituric acid reactive substance (TBARS) assay in porcine retina, and was found to decrease the production of TBARS. Electron spin resonance (ESR) measurements showed that NSP-116 exhibited radical scavenging activities against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, superoxide anion radical (center dot O-2(-)), and hydroxyl radical (center dot OH). These findings suggest that NSP-116 has protective effects against light-induced photoreceptor degeneration in vitro and in vivo as a free radical scavenger, and it may be a novel therapeutic agent for retinal degenerative disorders, such as dry age related macular degeneration (AMD). (C) 2016 Elsevier Inc. All rights reserved.