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

1.为了研究猪视网膜杆状外段(ROS)脂褐素的化学组成，根据脂质过氧化和蛋白质分解形成的醛可以产生脂褐素类物质的原理，将视网膜匀浆与FeCl2和白蛋白孵育，生成类脂褐素荧光团。用高效毛细管电泳法对孵育后视网膜匀浆中的ROS衍生荧光团进行了各种化学分析1)醛(HNE)，HNE在孵育初期有一个峰值，并随时间的延长而降低。显示脂质过氧化的TBARs在孵化早期增加，并在孵育24小时内保持平台期。随着时间的延长，荧光素强度逐渐增强。2)HNE和白蛋白孵育产生的荧光团的荧光光谱与视网膜匀浆与FeCl2和白蛋白孵育产生的荧光团的荧光光谱相对应。3)谷胱甘肽抑制本实验系统中的荧光团，因为谷胱甘肽可以与醛形成偶联物。铁诱导的过氧化处理的牛RPE吞噬的ROS比未处理的吞噬ROS的ROS显示出更强的荧光团。

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.