Bilberry and its main constituents have neuroprotective effects against retinal neuronal damage in vitro and in vivo

Abstract

Our aim was to determine whether a Vaccinium myrtillus (bilberry) anthocyanoside (VMA) and/or its main anthocyanidin constituents (cyanidin, delphinidin, and malvidin) can protect retinal ganglion cells (RGCs) against retinal damage in vitro and in vivo. In RGC cultures (RGC-5, a rat ganglion cell-line transformed using E1A virus) in vitro, cell damage and radical activation were induced by 3-(4-morpholinyl) sydnonimine hydrochloride (SIN-1, a peroxynitrite donor). Cell viability was measured using a water-soluble tetrazolium salt assay. Intracellular radical activation within RGC-5 cells was evaluated using 5-(and-6)-chloromethyl-2,7-dichlorodihydrofluorescein diacetate acetyl ester (CM-H(2)DCFDA). Lipid peroxidation was assessed using the supernatant fraction of mouse forebrain homogenates. In mice in vivo, we evaluated the effects of VMA on N-methyl-D-aspartic acid (NMDA)-induced retinal damage using hematoxylin-eosin and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) stainings. VMA and all three anthocyanidins (i) significantly inhibited SIN-1-induced neurotoxicity and radical activation in RGC-5, (ii) concentration-dependently inhibited lipid peroxidation in mouse forebrain homogenates. Intravitreously injected VMA significantly inhibited the NMDA-induced morphological retinal damage and increase in TUNEL-positive cells in the ganglion cell layer. Thus, VMA and its anthocyanidins have neuroprotective effects (exerted at least in part via an anti-oxidation mechanism) in these in vitro and in vivo models of retinal diseases.

Anthocyanins Extracted from Black Soybean Seed Coat Protect Primary Cortical Neurons against <i>in Vitro</i> Ischemia

“…9) Moreover, our results demonstrated that black soybean anthocyanins, including C3G, dose dependently inhibited excessive ROS generation in primary cortical neurons exposed to OGD, which coincided with increased cell viability. Our results parallel the finding that anthocyanins, including C3G, possess strong antioxidant capacities that are known to scavenge different free radicals, 35) inhibit lipid peroxidation, 36,37) and prevent free radical-induced PC12 cell death. 19) In addition, anthocyanins may modulate the activity of neuronal antioxidant enzymes.…”

“…However, there is an in vivo study that found that anthocyanins extracted from bilberry inhibited NMDA-induced retinal damage in mice. 37) It is reported that there are approximately 15 different anthocyanins in bilberry. 37,48) Therefore, it is possible that the anti-excitotoxic activity of bilberry extract may be due to the effect of anthocyanin components other than C3G, D3G, and P3G.…”

<i>In silico</i> docking studies of RP2 (X-Linked retinitis pigmentosa) protein using anthocyanins as potential inhibitors

“…Residues -Residues Leu 264, Ile 223, Ser 219, Arg 251, Asp 255, Ala 193, Gln 149 were found strongly interacting with the compound. An increased interest in cyanidin 3,5 diglucoside is due to its promising health benefits related to vision (Matsunaga et al,2009). Delphinidin 3,5 diglucoside generated a glide XP score of -12.55 kcal/mol, with a glide energy and glide emodel score of -67.25 kcal/mol and -92.73 kcal/mol respectively.…”

The Protective Effects of a Dietary Carotenoid, Astaxanthin, Against Light-Induced Retinal Damage

“…The 96-well plate was loaded onto a plate in a fluorescence spectrophotometer, and the reaction was carried out at 37°C. Cell fluorescence was determined by Hoechst 33342 staining and was used to calculate ROS production per cell (26).…”