Animals. For the evaluation of the biological activity of RLE, 9 to 12 CD1 male mice per experimental group, aged between 6 and 7 weeks, were used. All experiments were carried out in accordance with experimental protocols approved by the Italian Ministry of Health.

RLE administration. RLE was solubilized in water and mice received a daily oral administration of RLE containing 120 mg/kg/day−1 of total anthocyanins. The main individual compounds were cyanidin 3-glucoside, counting for 50 mg/kg/day−1, and cyanidin 3-(6-malonyl) glucoside), counting for 25 mg/kg/day−1. The total intake of flavanones amounted to 700 mg/kg/day−1 with the main individual compounds being eriocitrin (480 mg/kg/day−1), narirutin (60 mg/kg/day−1), hesperidin (150 mg/kg/day−1), and didymin (10 mg/kg/day−1).

High-fat diet. An adjusted calories diet was used for this study (Teklad Custom Research Diet, TD.06414) containing 60.3% of total calories from fat, 18.4% from proteins, and 21.3% from carbohydrates.

Evaluation of blood glucose, cholesterol, and triglycerides levels. Mice were fed on a high-fat diet (Harlan-DIETA TD 06414) for eight weeks and randomly divided into two groups. Concomitantly to the high-fat diet, one group received the RLE supplement (RLE-HFD mice) while the control group received water (Vehicle-HFD mice). For the evaluation of glucose, cholesterol, and triglycerides levels, 100 microliters of blood were drawn from the tail vein and a drop was put on multiCare in reactive strips (Biochemical Systems International). Results were expressed as mg/dl. Glucose, cholesterol, and triglycerides levels were evaluated before the high-fat diet administration (basal) or treatments and after 4 and 8 weeks of treatment. Glucose, cholesterol, and triglycerides blood levels were measured in mice before any treatment (basal) and after four and eight weeks of HFD.