"AGLIANICO DEL VULTURE" RED WINE POLYPHENOLIC EXTRACT (RWP) AS DIETARY SUPPLEMENT FOR PREVENTING OBESITY-ASSOCIATED CARDIOMETABOLIC COMPLICATIONS: THE IN VITRO AND IN VIVO PHARMACO-TOXICOLOGICAL PROFILE

Introduction. Evidence suggests that nutraceuticals containing polyphenols help to prevent atherosclerosis and cardiometabolic disease (CMD) complications by directly modulating signaling pathways in endothelial and inflammatory cells of the vascular compartment. Recently, components from the Aglianico del Vulture red wine (RWP) extract have demonstrated significant in vitro effects on polarization, histone acetylation, lipid-peroxidation and proinflammatory cytokine activities from human macrophages. Aim. To gain deeper insights on RWP vascular protective mechanisms, its pharmaco-toxicological profile has been evaluated on endothelial cells (ECs) from patients with CMD (obesity, dyslipidemia, and insulin resistance), and on spontaneously hypertensive rats (SHR), a metabolic syndrome animal model. Methods. ECs from CMD patients (n = 12) were incubated with RWP (dose-response: 0, 100, 200, 400, 800, 1600, 3200 lg/mL; timecourse: 0, 24, 48, 72 h) to measure toxicological (apoptosis, by flow cytometry) and functional (migration, by wound healing) effects. SHR 9 week (wk) old (n = 20) were randomized into 4 groups and treated (by gavage) with vehicle or RWP (200, 400, 800 mg/Kg/day) for 3 wk period. Body weight and food intake were measured daily, systolic blood pressure (SBP) every 3 days (tail cuff). At the end of the 3 wk treatment, plasma glucose concentrations and pro-inflammatory cytokine levels were measured. Vasodilation to acetylcholine (ACh, 10nM to 3lM/30 sec) and RWP (5 to 20 lg/ml/30 sec) was evaluated before and after N-nitro-L-arginine methyl ester preincubation (L-NAME, 100 lM/20 min) on mesenteric arteries (MVA) isolated and pre-constricted with noradrenaline (NA, 5lM). Results. On human ECs, RWP promoted apoptosis and impaired migration only at the highest doses (up to 800 lg/mL/72 h). Under in vivo studies, 3-wk treatment with RWP significantly reduced SBP levels in all groups (P\0.001, vs vehicle treated), with no associated signs of organ toxicity, or significant changes in body weight and blood glucose levels. Conversely, levels of pro-inflammatory cytokines as well as adhesion molecules tended to decrease. On ex vivo experiments, ACh-mediated MVA vasorelaxation was significantly increased in 400 and 800 mg/Kg/day RWP-SHR treated (vs vehicle-treated, p\0.05). Direct administration of RWP on MVA from 12-wk-old vehicle-treated SHR produced a reversible and dose dependent vasorelaxation abolished by L-NAME preincubation (P\0.001 vs respective control). Conclusions. Obtained data suggest that RWP contributes to cardiovascular protection via activation of NO-mediated signaling pathways and inhibition of NFkB-mediated inflammatory activities. Current experiments in co-coltured ECs and macrophages from CMD patients aim to clarify cellular mechanisms by which RWP administration may protect from initial atherosclerotic process.