Effects of organic solvents on two retinal pigment epithelial lipofuscin fluorophores, A2E and all-trans-retinal dimer  

有机溶剂对两种视网膜色素上皮脂褐质荧光色素的影响(英文)

作　　者：Qiu-xia JIN Xin-ran DONG Jing-meng CHEN Ke YAO Ya-lin WU 

机构地区：[1]College of Pharmaceutical Sciences, Zhejiang University [2]Eye Center, the Second Affiliated Hospital, School of Medicine, Zhejiang University [3]School of Medicine, Zhejiang University City College

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2014年第7期661-669,共9页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：Project supported by the National Natural Science Foundation of China(Nos.21202146 and 81271018);the Fundamental Research Funds for the Central Universities;the Zhejiang Scientific Research Foundation for the Returned Overseas Chinese Scholars(No.J20120556);the Zhejiang Key Laboratory Fund of China(No.2011E10006)

摘　　要：Gene and drug therapies are being developed to al eviate vision loss in patients with Stargardt’s disease and age-related macular degeneration （AMD）. To evaluate the therapeutic effects of these treatments, organic sol-vents 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 fluoro-phores in related experiments.研究目的:减缓Stargardt和年龄相关性黄斑变性患者视力损失的基因和药物疗法正处于发展中,然而为了评价对实验小鼠的治疗效果,有机溶剂是获取及定量小鼠后眼罩脂褐质色素必需的载体。本工作报道了有机溶剂对N-亚视黄基-N-视黄基乙醇胺(A2E)和全反式视黄醛二聚体两种重要脂褐质色素的影响以及从体内获取和定量它们的优化方法。创新要点:本文阐释了二甲亚砜、甲醇、氯仿、四氢呋喃、环己烷和乙醇对A2E和全反式视黄醛二聚体两种脂褐质色素的影响,并且优化了它们的体内获取及定量程序。研究方法:采用高效液相色谱技术分析A2E和全反式视黄醛二聚体溶液以及C57BL/6小鼠后眼罩浸膏。重要结论:氯仿和四氢呋喃能够改变A2E和全反式视黄醛二聚体,但是它们在二甲亚砜和甲醇中是稳定的;环己烷和乙醇不影响全反式视黄醛二聚体,然而A2E在乙醇中不稳定,在环己烷中溶解度很低。脂褐质元素体内获取及定量的优化方法能够释放出更多的全反式视黄醛二聚体和A2E。理解有机溶剂对脂褐质色素的影响将为正确使用这些色素提供洞察力,也可促进建立每个内源性色素测量的合理方法。

关 键 词：N-retinylidene-N-retinyl-ethanolamine （A2E） All-trans-retinal dimer LIPOFUSCIN Organic solvents Retinal pigment epithelial 

分 类 号：R774.1[医药卫生—眼科]