Effect of Aspirin on human ARPE-19 cells and in Mouse Model of Choroidal Neovascularization

Abstract

 

Purpose

 

Age-related macular degeneration (AMD) is a leading cause of irreversible blindness worldwide. Conflicting associations between aspirin (ASA) use and AMD have been observed in various epidemiologic studies questioning the safety of ASA in patients with concurrent AMD and heart disease. The aim of this study was to characterize the effect of aspirin administration on RPE cells in-vitro and in a mouse model of wet ARMD.

 

Methods

 

In- Vitro: Human ARPE-19 cells (obtained from ATCC) were cultured for 24 hours before applying 0.5 mM/L or 2.0 mM/L of ASA for 72 hours. ELISA was used to measure the concentration of VEGF, PDGF-BB and CCL-2 in the supernatants collected after 24 and 72 hours. In- Vivo: Male C57BL/6 mice (from Jackson laboratory) were given ASA (370 μg/mL) in drinking water daily for 7 days before and after laser treatment. CNV was induced by argon laser photocoagulation. Mice were sacrificed and perfused with PBS containing 50 mg/ml fluorescein isothiocyanate (FITC) conjugated dextran on day 7 post-laser. RPE-choroid-sclera complexes were stained for VEGF and F-actin using immunohistochemistry (IHC). The relative area of FITC-dextran perfused vessels, laser spot area and mean intensity of VEGF positive IHC staining in the laser injured zones was measured with ImagePro Plus software.

 

Results

 

ASA significantly reduced secretion of VEGF and CCL2 by human ARPE-19 cells in dose dependent manner. Though in-vivo ASA treatment showed some trend towards decrease in laser spot size, no statistically significant effect on either CNV size or laser spot size was noted. Similarly there was no statistically significant difference in levels of VEGF expression in ASA treated mice.

 

Conclusions

 

ASA may act as anti-angiogenic agent at higher doses as demonstrated by in-vitro study. Because of the prevalence variability of responsiveness of ASA in humans, it may be assumed that ASA at doses consumed clinically for various medical causes may not worsen CNV in human subjects. In- fact, ASA may have anti-angiogenic effects at higher doses.

 

 

Effect of Aspirin on CNV size and size of laser spot in mouse model of CNV

 

Keywords: 412 age-related macular degeneration