<p>Inflammatory bowel diseases are characterized by oxidative stress, microbial imbalance, and disruption of intestinal barrier integrity, highlighting a need for safe natural interventions. This study aimed to investigate the antioxidant, antibacterial, and anti-inflammatory properties of polysaccharides extracted from red beetroot (<i>Beta vulgaris</i> L., PSB) and to evaluate their potential to preserve intestinal epithelial barrier function through an in vitro model of inflammation. The antioxidant activity of PSB was measured using free radical scavenging and ferric reducing assays, while antibacterial activity was tested against selected intestinal pathogenic strains. To mimic intestinal inflammation, differentiated human intestinal epithelial cells (Caco-2) were co-cultured with macrophage-like immune cells (THP-1), and inflammation was induced by lipopolysaccharide (LPS) treatment. PSB were administered at increasing concentrations (100, 200, and 400&#xa0;µg/mL). Inflammatory mediators, oxidative stress markers, and transepithelial electrical resistance (TEER) were evaluated. The results showed that PSB demonstrated strong antioxidant potential and moderate antibacterial effects. In the inflamed co-culture system, PSB significantly reduced the secretion of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6), with a significant decrease of malondialdehyde (MDA) accumulation and the restoration of superoxide dismutase (SOD) activity. Additionally, PSB had minimal influence on epithelial barrier integrity under basal conditions; however, they preserved barrier function during inflammation, as shown by improved TEER compared with LPS-challenged controls. These findings highlight that PSB represent a promising natural strategy for the prevention and management of inflammatory bowel disorders.</p>