Master of Science (MS)
School of Nutrition and Food Sciences
This research investigated the ability of blueberry anthocyanins to inhibit the uptake of N-retinyl-N-retinylidene ethanolamine (A2E) by retinal pigment epithelial (RPE) cells as quantified by pigment epithelial derived factor (PEDF) levels in RPE cells. The A2E was synthesized and identified by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Freeze dried blueberry powder of 50/50 blend (Tifblue/Rubel) anthocyanins were extracted by acetone and concentrated through chromatography. A RPE cell culture study investigating four treatments using a combination of 100 ìM A2E in DMSO and 100 ìg of blueberry anthocyanins in DMSO was tested. The treatments included: (A) control RPE cells; (B) RPE cells incubated with A2E for 2 hours with blue light illumination for 20 minutes; (C) RPE cells incubated with blueberry anthocyanins for 24 hours and incubated with A2E for 2 hours with blue light illumination for 20 minutes; and (D) RPE cells incubated with blueberry anthocyanins for 24 hours, then incubated with A2E for 2 hours with blue light illumination for 20 minutes and then treated with blueberry anthocyanins again. All treatments were incubated for seven days at 37°C with 5% CO2 and samples were collected on days 1, 3, 5 and 7. The supernatant of the RPE cells were collected and the protein concentration determined and normalized. The PEDF values were determined using a PEDF enzyme linked immunosorbant assay (ELISA). The average PEDF results indicated that blueberry anthocyanins promoted and maintained PEDF levels when compared to control RPE cells and RPE cells treated with A2E for 1, 3 and 5 collection days. Treatments C and D had significantly (p<0.05) higher PEDF levels of 10.17 and 10.14 ng PEDF per mg protein than treatment B of 5.750 ng PEDF per mg protein for day 1. Treatment D had a significantly (p<0.05) higher PEDF level of 13.76 ng PEDF per mg protein than the other treatments at day 3 and was significantly higher at 12.51 ng PEDF per mg protein than treatment A at 8.947 ng PEDF per mg protein on collection day 5. There is evidence that blueberry anthocyanins protected RPE cells from oxidation as measured by PEDF values.
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Sundalius, Naomi Marie, "Examination of blueberry anthocyanins in prevention of age-related macular degeneration through retinal pigment epithelial cell culture study" (2008). LSU Master's Theses. 974.