Abstract
Purpose :
Age-related macular degeneration (AMD) is a multifactorial retinal disease. Amyloid-β (Aβ) deposition has been reported in patients with AMD among the constituents of drusen, which cause inflammation, oxidative stress and retinal pigmented epithelium (RPE) damage. Hereby, we assessed the effect of vitamin D3 and meso-zeaxanthin combination in several in-vitro models of AMD, challenging retinal cells with three different types of damage.
Methods :
Human ARPE-19 cells were exposed to three different stimuli (Aβ, LPS, H2O2) in separate set of experiments. For all experiments, the cells were pre-treated for 24 hours with vitamin D3 (50 nM-1 µM), meso-zeaxanthin (0.1 µM and 0.5 µM) or their combination; then, ARPE-19 were exposed to the different insults in separate set of experiments. The damage of Aβ, LPS, H2O2, and the protective effects of molecules were evaluated with MTT, LDH, ROS assay and the modulation of inflammatory mediators (IL-1β, IL-6, TNFα, MMP-9 and VEGF) was evaluated with RT-PCR.
Results :
The combination of vitamin D3 and meso-zeaxanthin significantly protected ARPE-19 against the damage elicited by Aβ, LPS, and H2O2. These three stimuli significantly (p<0.05) reduced cell proliferation, increased LDH and ROS release after different time of treatment. These effects were significantly (p<0.05) counteracted by the combination of vitamin D3 and meso-zeaxanthin treatment. Further, the expression of inflammatory mediators such as IL-1β, IL-6 and TNFα was significantly (p<0.05) reduced by vitamin D3 and meso-zeaxanthin combination treatment. Finally, a potentiation effect has been demonstrated by vitamin D3/meso-zeaxanthin; the combination was, in fact, able to reduce significantly (p<0.05) the expression of VEGF-A and MMP-9 mRNA, in ARPE-19 challenged with LPS and H2O2 in comparison with the single molecules
Conclusions :
These findings demonstrated, at least in these in-vitro paradigms, that combination of vitamin D3/meso-zeaxanthin exerts an enhanced protective effect in ARPE-19 cells damaged by amyloid-β, inflammation (LPS) and oxidative stress (H2O2) that represent the initiating events leading to AMD.
This is a 2021 ARVO Annual Meeting abstract.