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(Investigative Ophthalmology and Visual Science. 2007;48:2343-2349.)
© 2007 by The Association for Research in Vision and Ophthalmology, Inc.
doi:10.1167/iovs.06-0452
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Bilberry (Vaccinium myrtillus) Anthocyanins Modulate Heme Oxygenase-1 and Glutathione S-Transferase-pi Expression in ARPE-19 Cells

Paul E. Milbury,1 Brigitte Graf,2 Joanne M. Curran-Celentano,3 and Jeffrey B. Blumberg1

1From the Antioxidants Research Laboratory, Jean Mayer USDA Human Nutrition Research Center on Aging, Tufts University, Boston, Massachusetts; and the 3Department of Animal and Nutritional Sciences, University of New Hampshire, Durham, New Hampshire.

PURPOSE. To determine whether anthocyanin-enriched bilberry extracts modulate pre- or posttranslational levels of oxidative stress defense enzymes heme-oxygenase (HO)-1 and glutathione S-transferase-pi (GST-pi) in cultured human retinal pigment epithelial (RPE) cells.

METHODS. Confluent ARPE-19 cells were preincubated with anthocyanin and nonanthocyanin phenolic fractions of a 25% enriched extract of bilberry (10–6–1.0 mg/mL) and, after phenolic removal, cells were oxidatively challenged with H2O2. The concentration of intracellular glutathione was measured by HPLC and free radical production determined by the dichlorofluorescin diacetate assay. HO-1 and GST-pi protein and mRNA levels were determined by Western blot and RT-PCR, respectively.

RESULTS. Preincubation with bilberry extract ameliorated the intracellular increase of H2O2-induced free radicals in RPE, though H2O2 cytotoxicity was not affected. By 4 hours, the extract had upregulated HO-1 and GST-pi protein by 2.8- and 2.5-fold, respectively, and mRNA by 5.5- and 7.1-fold, respectively, in a dose-dependent manner. Anthocyanin and nonanthocyanin phenolic fractions contributed similarly to mRNA upregulation.

CONCLUSIONS. Anthocyanins and other phenolics from bilberry upregulate the oxidative stress defense enzymes HO-1 and GST-pi in RPE, suggesting that they stimulate signal transduction pathways influencing genes controlled by the antioxidant response element.




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J. Johnson, P. Maher, and A. Hanneken
The Flavonoid, Eriodictyol, Induces Long-term Protection in ARPE-19 Cells through Its Effects on Nrf2 Activation and Phase 2 Gene Expression
Invest. Ophthalmol. Vis. Sci., May 1, 2009; 50(5): 2398 - 2406.
[Abstract] [Full Text] [PDF]


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