Effects of organic solvents on two retinal pigment epithelial lipofuscin fluorophores, A2E and all-trans-retinal dimer
Effects of organic solvents on two retinal pigment epithelial lipofuscin fluorophores, A2E and all-trans-retinal dimer
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DOI：
10.1631/jzus.b1300194
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发表时间：
2014-07
影响因子：
5.1
通讯作者：
Qiu-xia Jin;Xinran Dong;Jing-meng Chen;Ke Yao;Yalin Wu
中科院分区：
文献类型：
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作者：
Qiu-xia Jin;Xinran Dong;Jing-meng Chen;Ke Yao;Yalin Wu
目前正在开发基因和药物疗法，以减轻Stargardt病和年龄相关性黄斑变性（AMD）患者的视力丧失。为了评价这些治疗的治疗效果，常规使用有机溶剂来提取和定量双视黄脂褐素成分，例如N-亚视黄基-N-视黄基-乙醇胺（A2 E）和全反式-视黄二聚体（ATR-二聚体）。通过高效液相色谱（HPLC）分析，发现A2 E和ATR-二聚体在四氢呋喃（THF）和氯仿中均发生变化，但在二甲基亚砜（DMSO）和甲醇（MeOH）中稳定。此外，环己烷和乙醇（EtOH）没有改变ATR-二聚体，而在EtOH中发生A2 E的改变。在这些发现的基础上，我们设计了过程II-IV，通过过程I的修改产生，这是一种测量体内bisretinoid化合物的常规方法。与过程I相比，过程II-IV在小鼠眼杯中释放了额外量的ATR-二聚体或A2 E。努力澄清有机溶剂对脂褐素色素的影响是重要的，因为这样的研究可以指导在相关实验中处理这些荧光团。
Gene and drug therapies are being developed to alleviate vision loss in patients with Stargardt’s disease and age-related macular degeneration (AMD). To evaluate the therapeutic effects of these treatments, organic solvents are routinely used to extract and quantify bisretinoid lipofuscin constituents, such as N-retinylidene-N-retinyl-ethanolamine (A2E) and all-trans-retinal dimer (ATR-dimer). By high-performance liquid chromatography (HPLC), we found that A2E and ATR-dimer were both altered by tetrahydrofuran (THF) and chloroform, but were stable in dimethyl sulfoxide (DMSO) or methanol (MeOH). In addition, cyclohexane and ethanol (EtOH) did not alter ATR-dimer, whereas an alteration of A2E occurred in EtOH. On the basis of these findings, we designed processes II–IV, generated by modifications of process I, a routine method to measure bisretinoid compounds in vivo. Extra amounts of either ATR-dimer or A2E in mouse eyecups were released by processes II–IV versus process I. Efforts to clarify the effects of organic solvents on lipofuscin pigments are important because such studies can guide the handling of these fluorophores in related experiments.