Abstract
Purpose :
Impaired autophagic and proteasomal cleansing has been documented in retinal pigment epithelial (RPE) cells and age-related macular degeneration (AMD) pathology. Omega fatty acids and resveratrol has been shown to be cytoprotective in RPE cells. We examined effects of commercial Resvega on the regulation of autophagy and cytoprotection in ARPE-19 cells under proteasome inhibition.
Methods :
Protein aggregation was induced with 1 µM MG-132. Resvega, which includes vitamin C 240 mg, E 30 mg, zinc 12,5 mg, copper 1 mg, omega-3 665 mg, lutein 10 mg, zeaxanthin 2 mg and resveratrol 30 mg, was used solely and together with proteasome inhibition up to 48 hours.. SQSTM1/p62, MAP1LC3A/LC3, ubiquitin protein conjugates and Hsp70 were analyzed by western blotting (WB). Tandem fluorescently tagged LC3 (GFP-mCherry-LC3) transfection was used to study autophagy flux in fluorescence microscopy.
Results :
Inhibition of proteasomes with MG-132 upregulated ubiquitin protein conjugates and Hsp70 and autophagy markers SQSTM1/p62 and MAP1LC3A/LC3 detected in WB. Simultaneous treatment with MG-132 and Resvega corresponding 15 and 25 µM resveratrol concentration highly increased amount of LC3-II, but decreased SQSTM1/p62 and Hsp70. Moreover, Resvega provided a clear cytoprotection under proteasome inhibition. Fluorescence microscopy showed increased autophagy flux with GFP-mCherry-LC3 Resvega treatment.
Conclusions :
Resvega has a potential to further studies in the prevention of RPE damage and AMD.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.