Abstract: This study used goat milk oligosaccharides (GMOs) isolated from goat milk and Bifidobacterium bifidum ATCC15696 (BB) as raw materials to construct a Caco-2:HT-29MIX intestinal epithelial barrier and inflammatory injury model to explore the individual MOs and BB and their synergistic effects on the integrity and permeability of the intestinal barrier and the expression levels of inflammatory factors 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, and the recovery effect on barrier function was the best and the expression of inflammatory factors was significantly inhibited when the action concentration is 10.0 mg/mL.