The effect of α tocopherol, all-trans retinol and retinyl palmitate on the non enzymatic lipid peroxidation of rod outer segments

The effect of alpha tocopherol, all-trans retinol and retinyl palmitate on the non enzymatic lipid peroxidation induced by ascorbate-Fe2+ of rod outer segment membranes isolated from bovine retina was examined. The inhibition of light emission (maximal induced chemiluminescence) by alpha tocopherol, all-trans retinol and retinyl palmitate was concentration dependent. All trans retinol showed a substantial degree of inhibition against ascorbate-Fe2+ induced lipid peroxidation in rod outer segment membranes that was 10 times higher than the observed in the presence of either alpha tocopherol or retinyl palmitate. Inhibition of lipid peroxidation of rod outer segment membranes by alpha tocopherol and retinyl palmitate was almost linear for up to 0,5 mu mol vitamin/mg membrane protein, whereas all-trans retinol showed linearity up to 0,1 mu mol vitamin/mg membrane protein. Incubation of rod outer segments with increasing amounts of low molecular weight cytosolic proteins carrying 1-[C-14] linoleic acid, [H-3] retinyl palmitate or [H-3] all-trans retinol during the lipid peroxidation process produced a net transfer of ligand from soluble protein to membranes. Linoleic acid was 4 times more effectively transferred to rod outer segment membranes than all-trans retinol or retinyl palmitate. Incubation of rod outer segments with delipidated low molecular weight cytosolic proteins produced inhibition of lipid peroxidation. The inhibitory effect was increased when the soluble retinal protein fraction containing a tocopherol was used. These data provide strong support for the role of all-trans retinol as the major retinal antioxidant and open the way for many fruitful studies on the interaction and precise roles of low molecular weight cytosolic retinal proteins involved in the binding of antioxidant hydrophobic compounds with rod outer segments.