Abstract: This study used goat milk oligosaccharides (GMOs) isolated from goat milk and Bifidobacterium bifidum ATCC15696 (BB) as raw materials. By constructing a Caco-2:HT-29MIX cell intestinal epithelial barrier and an inflammation injury model, the effects of GMOs alone, BB alone, and their synergistic interaction on intestinal barrier integrity, permeability, and the expression levels of inflammatory cytokines were investigated. The results showed that GMOs alone markedly increased transepithelial electrical resistance (TEER) in a dose-dependent manner, inhibited the secretion of inflammatory factors and promoted the expression of tight junction proteins, and the effect was superior to that of BB. However, GMOs and BB acted in synergy, the effect was more significant (P<0.01). When the action concentration was 10.0 mg/mL, the change rate of the TEER increased by 1.62%~3.27% compared with the GMOs group (P<0.01). The expression levels of IL-6, IL-8, and TNF-α decreased (2.11~14.93 ng/L), (7.42~16.27 ng/L) and (7.03~12.29 ng/L) (P<0.05), respectively. The expression levels of the three tight junction proteins, claudin-1, ZO-1 and occludin, increased by 1.0~2.0 times (P<0.001). Overall, individual GMOs and BB could improve the intestinal barrier function and inhibit the secretion of inflammatory factors. However, the synergistic effect of GMOs and BB was more significant, with the best recovery effect on barrier function and a significant inhibition of inflammatory factor expression at a concentration of 10.0 mg/mL.