Formulações naturais seguras, saudáveis e sustentáveis à base de frutos vermelhos para a prevenção de doenças cardiovasculares

The aims of this research will be: 1. Evaluation of the anti-diabesity of red fruits, by-products and their mixtures by in vitro assays. 2. Development of different formulations from red fruits and by-products. Stability and chromatographic characterization in different encapsulation methods will be performed to determine their bioactive compounds and to choose the most promising ones. Additionally, a three-dimensional in vitro assay based on a collagen layer with human intestinal fibroblasts embedded and epithelial cells on top capable of mimicking the small intestine will be done to assess formulations bioavailability. 3. The antioxidant, anti-inflammatory, antidiabetic and anti-obesity effects of the most promising formulations will be evaluated. The antioxidant capacity will be tested against ROS and RNS. The anti-inflammatory effects will be assessed in RAW 264.7 macrophages. The antidiabetic effects will be based on the ability of the formulations to inhibit a-amylase and a-glucosidase and to interfere with insulin signalling, and glucose production and uptake in insulin-responsive human HepG2 cells treated with high glucose. The anti-obesity will be determined by the ability of the different formulations to inhibit pancreatic lipase. 4. Anti-proliferative and cytotoxicity activity of beverage formulations using several human cell lines will be evaluated. Glycolytic metabolism and glucose consumption will be also conducted. The capacity of these formulations to protect several cells against oxidative stress will be also tested, including the effects on enzymatic, non-enzymatic antioxidants, and in the upregulation of phase II detoxification. 5. The most promising formulations will also be characterized in respect to their chemical composition and sensorial characteristics, being also evaluated their useful life period. 6. In vivo assays in rodents in order to evaluate the protective effects of red fruits formulations against diabetes, and obesity will be assessed in streptozotocin-induced diabetic rodents, and in obesity-induced rodents, respectively, 2 control groups (1 negative and 1 positive) that will be fed normal food and more 5 groups, where different doses of the most promising beverage will be orally administrated in accordance with WHO and EMA recommendations. 7. Clinical trials based on the daily ingestion of the best functional formulation with 80 humans with risk factors for the development of CVDs (namely with diabetes, obesity of hypertension) will be conducted for 3 months. Blood, saliva and urine samples will be collected before, during, and after 3 months, and analysed by chromatographic and laboratory methods to understand what parameters are more affected after beverage intake. At the end of the project, the biological potential of red fruit formulations will be revealed. Cova da Beira region is considered a disadvantaged region due to low population density and low income per capita, this project also pretends to increase the value of red fruits, and the region presents an ideal microclimate to produce red fruits, being a good source of income, allowing the establishment of people and attracting investment to the region. During the project, we will be in permanent contact with Cerfundão, BeiraBerry, 7CBAFruit, Frulact, etc., and also with the INOVCLUSTER to promote this product's consumption at the national and international levels.