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

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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.