Introduction: Metabiotics are promising candidates for peptic ulcer (PU) healing by regulating the expression of pro-inflammatory genes and antioxidant capacity. Methods: We prepared a metabiotic by culturing Lactobacillus acidophilus BF-P-064 and evaluated its cytotoxicity and antioxidant properties by MTT and DPPH (α-diphenyl-βpicrylhydrazyl) methods. To investigate the potential of PU healing, we divided 45 rats into 3 groups and induced PU by administering ethanol. The negative-control group was not treated, while the positive-control and metabiotic groups received treatment with a drug and a metabiotic. On different days, the wound area was determined and the histopathological characteristics were evaluated by H&E staining. The expression of pro-inflammatory cytokines (IL-6, PDGF, VEGF, EGF, TGF-β, and TNF-α) and antioxidant (SOD and GPx) genes was assayed by qRT-PCR method. Results: Our results showed that the metabiotic, at a concentration of 60 mg/mL, had no cytotoxic effects on the CaCo2 cell line, and its antioxidant capacity was 72.49 u/mL. The metabiotic caused faster wound contraction by accelerating the infiltration of phagocytic cells, which intensified the inflammatory phase. By increasing the expression of proinflammatory cytokines, as well as enhancing antioxidant properties and the expression of SOD and GPx, the metabiotic prevented the induction of oxidative stress in the tissue. So, the proliferation phase started faster, and the healing process improved. Conclusion: The metabiotic extracted from L. acidophilus BF-P-064 can improve the healing process of PU by increasing the antioxidant capacity, improving the pattern of cell infiltration, and the expression of pro-inflammatory cytokine