The study on the chemical analysis of ROS derived lipofuscin and the related neovascular factor

1.为了研究猪视网膜视杆细胞外段（rod outer segment，ROS）脂褐素的化学组成，根据脂质过氧化物和蛋白质分解生成的醛类物质可产生脂褐素样物质的理论，将猪视网膜匀浆与FeCl_2和白蛋白共同孵育，产生脂褐素样荧光团。对ROS衍生荧光团进行各种化学分析：1）用高效液相色谱法分析孵育后视网膜匀浆中的醛（HNE），HNE在孵育初期有一个峰值，随着孵育时间的延长而下降。TBARS显示脂质过氧化物在孵育早期增加，在孵育24小时内保持稳定。HNE与白蛋白孵育产生的荧光团的荧光光谱与视网膜匀浆与FeCl_2和白蛋白孵育产生的荧光团的荧光光谱相对应; 3）谷胱甘肽可与谷胱甘肽形成结合物，从而抑制了本实验系统中的荧光团.牛RPE吞噬的活性氧与铁诱导的过氧化处理显示出更强的荧光团比吞噬的活性氧没有处理。

1. In order to study on the analysis of chemical composition of lipofuscin derived from rod outer segment (ROS) in the pig retina, the retina homogenates were incubated with FeCl_2 and albumin to produce lipofuscin-like fluorophores, based on the theory that aldehydes formed by the decomposition of lipid peroxide and protein can produce lipofuscin-like substance. The various kinds of chemical analysis on ROS derived fluorophores were performed.1) Aldehyde (HNE) in the retina homogenates after the incubation was analyzed by HPLC.HNE had a peak at the initial incubation and decreased with the time. TBARS showing lipid peroxides increased at early time of incubation, keeping a plateau for 24 hr incubation. Fluorescein intensiy increased gradually with the time.2) Fluorescence spectrum on the fluorophores produced by the incubation of HNE and albumin corresponded to that from the fluophores produced by the incubation of the retina homogenates with FeCl_2 and albumin.3) Glutathione inhibited the fluorophores in this experimental sistem because glutathion can forms the conjugate with the aldehyde.2. The bovine RPE phagocyted ROS treated with the iron-induced peroxidation showed more intense fluorophores than that phagocyted ROS without the treatment.