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Nasim Salimiagdam, Marilyn Chwa, Anthony Nesburn, M.Cristina Kenney; Effects of Quercetin in AMD Cybrid Cell Lines: Implications for a Therapeutic Role in AMD. Invest. Ophthalmol. Vis. Sci. 2022;63(7):467 – A0004.
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© ARVO (1962-2015); The Authors (2016-present)
We evaluated the possible beneficial outcomes of quercetin on human AMD cybrid cell lines.IntroductionIn developed countries, age related macular degeneration (AMD) is the leading cause of progressive central vision loss in aged populations. Previous studies have demonstrated the advantageous impacts of antioxidants such as vitamin E and zinc in lowering the incidence and progression of AMD. Quercetin, a plant flavanol from the flavonoid group of polyphenols, is found in many fruits, vegetables and seeds. Previous in vitro studies have demonstrated the antioxidant and anti-inflammatory properties of quercetin, suggesting it may have therapeutic benefit for human diseases. Its main antioxidant activity is associated with inhibition of the signal transduction pathway and activity of cytosolic glutathione, leading to reduction of reactive oxygen species (ROS) production.
Cybrids were created by fusion of the mitochondria-deficient (Rho0) ARPE-19 cells with the platelets derived from AMD patients and age-matched non-AMD individuals. The resultant cybrid cell lines have mitochondria originating from different individuals but identical nuclei. Cells were cultured in 96-well plates (10,000 cells/well) and treated with quercetin 20 µM for 24 hours. DMSO was used as vehicle-control. ROS levels and cellular metabolism were measured in control and treated cultures. For ROS assay, after adding 100 µl H2DCFDA solution/well, plates were read via the fluorescent plate reader with the excitation (EX, 492nm) and emission (EM, 520nm) wavelengths. Performing MTT assay, 10 µl of MTT assay reagent was added/well and plates incubated in 37°C for 3 hours. After adding 200 μl/well of DMSO, plates were analyzed (signal at 570 nm and reference at 630 nm) with the absorbance plate reader. Statistical analyses were performed via student t-test.
ROS levels of quercetin treated non-AMD cybrids and AMD cybrids declined to 84.25% (P= 0.0002, n=4) and 86.75% (P= 0.011, n=4), respectively, compared to vehicle-control groups (100%, n=4). Moreover, quercetin increased cellular metabolism of non-AMD cybrids and AMD cybrids to 132.8% (P= 0.002, n=4) and 128.0% (P= 0.001, n=4), in order, versus vehicle-control cells (100%, n=4).
Administration of quercetin 20 µM has positive impacts on treated AMD cybrid cell lines.
This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.
* P<0.05 ** P<0.01 *** P<0.0001
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