Lingonberry Anthocyanins Protect Cardiac Cells from Oxidative Stress‐Induced Apoptosis

Lingonberry grown in Canada contains exceptionally high levels of anthocyanins, the pigments that give the berries their red color, as well as other polyphenols, including resveratrol, quercetin, and proanthocyanidins, which may have antioxidant properties. Anthocyanin‐rich diets have been shown to be associated with decreased cardiovascular disease and overall mortality. Oxidative stress plays an important role in cardiovascular disease by impairing function and triggering apoptosis of cardiomyocytes. In this study, we investigated whether physiologically relevant doses of pure anthocyanins and lingonberry extracts could protect H9c2 cardiomyoblasts from oxidative stress‐induced cell death. Anthocyanins were added at 10 ng mL −1 (20 nmol L −1 ) to cell culture media prior to induction of oxidative stress with 600 μM hydrogen peroxide. After 2 hours of hydrogen peroxide treatment, apoptosis and necrosis were detected in H9c2 cells via flow cytometry using FLICA 660 caspase 3/7 combined with YO‐PRO‐1 to allow differentiation between viable, early apoptotic, late apoptotic, and necrotic cells. Apoptosis was confirmed using Hoechst 33258 and fluorescence microscopy. Each of the three major anthocyanins found in lingonberry, cyanidin‐3‐galactoside, cyandin‐3‐glucoside, and cyanidin‐3‐arabinoside, was protective against hydrogen peroxide‐induced apoptosis in H9c2 cells at 10 ng mL −1 and preserved the number of viable cells to match the non‐hydrogen peroxide‐treated group. A combination of the three anthocyanins tested at 10 ng mL −1 was also protective. A lingonberry extract tested at three concentrations produced a dose‐dependent protective effect. This study demonstrates that lingonberry anthocyanins could protect cardiac cells from oxidative stress‐induced apoptosis and may have cardioprotective effects. Support or Funding Information AAFC Project ID J‐001297