BackgroundOwing to the emergence and spread of multidrug resistance mechanisms inHelicobacter pylori, achieving a successful eradication has become exceedingly difficult. Thus, this study for the first time determines the effect of a combination of vitamin D3 and probiotic on the pathogenesis and treatment ofH. pylori.MethodsWe established anin vitroexperimental system using AGS human gastric carcinoma cells and explored the synergistic effect ofLevilactobacillus brevisIBRC-M10790 and vitamin D3 onH. pylori. Live and pasteurizedL. brevis,L. brevis-derived membrane vesicles (MVs), andL. breviscell-free supernatant (CFS), as well as their combination with vitamin D3 were used during this study. We assessed the anti-inflammatory and anti-oxidative effects of these combinations using RT-qPCR and ELISA, respectively. We further performed an adhesion assay to evaluate the influence ofL. brevisand vitamin D3 on the adherence rate ofH. pylorito AGS cells.ResultsOur results demonstrated thatL. brevisand vitamin D3 possess anti-inflammatory and anti-oxidative effects againstH. pyloriinfection in AGS cells. The combination of vitamin D3 with the probiotic strain (particularly liveL. brevisand its CFS) can more efficiently reduce the expression of pro-inflammatory cytokines IL-6, IL-8, IFN-γ, and TNF-α in the AGS cells. Moreover, vitamin D3 andL. brevisexhibited an additive impact preserving the integrity of the epithelial barrier by increasing the expression of the tight junction protein ZO-1. Furthermore, this combination can potentially reduceH. pyloriadherence to AGS cells.ConclusionsThis study indicates the advantage of combining vitamin D3 and probiotic to attenuateH. pylori-induced inflammation and oxidative stress. Consequently, probiotic and vitamin D3 co-supplementation can be considered as a novel therapeutic approach to manage and preventH. pyloriinfection.