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Increased Expression of Vascular Endothelial Growth-factor (VEGF) Associated with the Accumulation of Lipids in Bruch`s Membrane of LDL-receptor-knock-out (LDLr(-))-Mice
1Rudolf M., 2Ivandic B. T., 2Winkler J., 2Schmidt-Erfurth U.,
1 (Lübeck)
2Medizinische Universität zu Lübeck, Klinik für Augenheilkunde (Lübeck)
Purpose: To investigate the pathogenesis of neovascular age-related macular degeneration (AMD) with respect to lipid accumulation within Bruch`s membrane in a knock-out model with low-density lipoprotein (LDL) receptor deficiency.
Methods: 4-months-old LDLr(-) mice and C57BL/6 controls were fed a standard rodent diet or a high fat diet western type for two months. Animals were sacrificed and serum cholesterol levels were determined. Eyes were examined by transmission electron microscopy (TEM). Immunohistochemical staining using monoclonal antibodies against VEGF was performed.
Results: High fat diet produced serum total cholesterol levels ranging from 281-702 mg/dl in LDLr(-) mice and from 146-215 mg/dl in controls. While Bruch`s membranes of control animals did not exhibit any visible changes by TEM even after fat diet, membrane-bound translucent particles were seen in Bruch´s membrane of all LDLr(-) eyes. The amount of these particles was substantially increased and membranes were thickened in LDLr(-) mice following high fat diet with additional deposits of non-membrane-bound particles. VEGF staining was positive in LDLr(-) mice only and was located in the outer plexiform layer and photoreceptor inner segments. Most intensive VEGF expression was documented in LDLr(-) mice following high fat diet corresponding to the increased amount of basal laminar deposits.
Conclusions: LDLr(-) mice exhibited an accumulation of lipid particles within Bruch`s membrane which is further increased after fat intake. VEGF expression is found within the outer retinal layers of LDLr(-) mice and appears to correlate with the amount of lipid particles present in Bruch`s membrane. Lipid deposits may stimulate VEGF expression and may be responsible for the progression of early AMD to advanced neovascular stages.