Abstract
Background: The intestine functions as the body’s primary immune barrier, playing a vital role in host defense against pathogens. Human milk oligosaccharides (HMOs: 2′-fucosyllactose and lacto-N-neotetraose) and α-milk fat globule membrane (α-MFGM) are key bioactive components in human milk known to regulate intestinal immunity. Although their individual benefits have been reported, the potential combined effects of their combined use on intestinal mucosal and systemic immune function remain poorly understood. Methods: To address this, we established a lipopolysaccharide (LPS)-induced intestinal injury model in BALB/c mice. The animals were randomly divided into five groups: normal control, LPS model control, α-MFGM alone, HMOs alone (2′-FL + LNnT), and combined α-MFGM + HMOs intervention. After 15 days of oral gavage, colonic histopathology, intestinal barrier integrity, immune organ indices, serum inflammatory cytokine levels, immune cell subsets, and gut microbiota composition were comprehensively evaluated. Results: Combined supplementation with HMOs and α-MFGM effectively alleviated colonic pathological damage, increased goblet cell numbers and mucus secretion, enhanced sIgA expression, and reduced serum DAO and D-lactate levels, indicating restored intestinal barrier function. It also balanced spleen and thymus indices, suppressed pro-inflammatory TNF-α/IL-6 production, elevated anti-inflammatory IL-10/IL-22 levels, and corrected the LPS-induced Th1/Th2 and Th17/Treg imbalances. Moreover, the combined intervention remodeled the gut microbiota by enriching beneficial Lactobacillus and Roseburia while decreasing the abundance of Prevotella abundance, with functional predictions pointing to enhanced butanoate metabolism. Conclusions: These findings demonstrate that HMOs and α-MFGM exert combined protective effects against LPS-induced intestinal immune dysfunction through modulation of the gut microbiota and its metabolic products, providing a scientific foundation for optimizing early-life nutritional strategies.