[Objective] Investigating the relationship between Lactobacillus animalis LGM and transcription factors (T-bet, GATA3, ROR-γt and Foxp3) gene expressions in vivo or in vitro and observing the effects of L. animalis LGM gavage on DSS-induced colitis.[Methods] L. animalis LGM was isolated from pig colon contents using Hungate rolling tube technique. Bacterial supernatants were extracted and incubated Caco-2 cells with LPS (2 μg/mL) simultaneously. Bacterial suspension was adjusted to 1×10⁹ CFU/0.2 mL and gavaged to DSS-induced mice. Gene expressions of transcription factors (T-bet, GATA-3, ROR-γt and Foxp3) and intracellular cytokines (IL-4, IL-17, INF-γ and IL-10) were analyzed by real-time PCR. [Results] In Caco-2 cells culture experiment, compared to control and LPS groups, L. animalis LGM supernatants treatment significantly downregulated the GATA-3, IL-4, IL-17 and TGF-β gene expressions and upregulated Foxp3 and ROR-γt gene expressions (P<0.05). In DSS-induced colitis mice, compared to the DSS group, L. animalis LGM administration restored the colonic length and increased ROR-γt and Foxp3 gene expressions (P<0.05). [Conclusion] L. animalis LGM isolated from pig colon contents in the present study have distinctive effects on transcription factors. ROR-γt and Foxp3 expressions were significantly upregulated both in Caco-2 cells and DSS-induced colitis mice. L. animalis LGM showed protection on DSS-induced colitis and gut inflammation.