Aspirin and its Metabolites Enhance the Expression of Vascular Endothelial Growth Factor in Retinal Pigment Epithelial Cell Cultures ? Implications in the Pathophysiology of Age-Related Macular Degeneration

Purpose: An estimated 19.3% of adults, especially the elderly in the United States regularly use Aspirin for cardioprotection. Recently, multiple cohort studies have concluded that regular aspirin use for 10-15 years was associated with a statistically significant increase in the risk of incident age-related and neovascular acute macular degeneration. It has been hypothesized that aspirin or its metabolites induce the expression of vascular endothelial growth factor (VEGF). Materials & Methods: Retinal pigment epithelial cells, ARPE-19 (ATCC®CRL-2302™) were cultured. The cells were grown to achieve 95% confluence and then the media was changed. Cells cultured under blue light, red light, or darkness were subjected to a challenge with high dose aspirin (0.925 mg/dL), low dose aspirin (0.325 mg/dL), or hippuric acid (0.325 mg/dL). Light was generated using 2 red or blue LEDs powered by 3v CR2032 batteries. The 24-well plate was incubated with or without drugs in blue light, red light or darkness at 37C for 16 hours. The supernatants were harvested, and VEGF was quantified. One-way ANOVA using Dunnett’s multiple comparison test was performed to analyze statistical significance. Results: Cells exposed to blue light or darkness and hippuric acid showed a statistically significant increase in VEGF secretion (P=0.0012). However, cells exposed to red light with hippuric acid challenge showed no significant difference from the mean of cells exposed to darkness and sham control. Conclusions: Retinal pigment epithelial cells challenged with oxidative stress provided by blue light or darkness in the presence of hippuric acid increased VEGF secretion, suggesting a possible cause for neovascularization in age-related macular degeneration. RPE cells exposed to red light, known to abrogate oxidative stress, had decreased levels of VEGF induction by hippuric acid.