Objective The aim of this study was to investigate how different concentrations of dextran sulfate sodium (DSS) influence the establishment of mouse model of inflammatory bowel disease (IBD) and the effect of DSS on the expression of colitis-associated immune factors. Methods The DSS solution in different concentrations (3%, 5%, 7%) were given to male C57BL/6J mice to generate mouse inflammatory bowel disease model. The IBD mice were observed by defecation characteristics, body weight, and survival time. The animals were sacrificed at 6 days after the start of DSS drinking. The general appearance of colons was observed and scored. Moreover, the pathological changes of the colon were examined and analyzed by routine histology. The expression of immune factors in the spleen was detected by real-time PCR.Results The mice in the 3%, 5%, 7% DSS groups developed murine colitis. In addition, the incidence of IBD and mouse mortality rate was directly proportional to the increase of DSS concentration. Furthermore, the higher concentration of DSS induced the expression of proinflammatory factors including TNF-α, IFN-γ and IL-17A, but cause a decrease of anti-inflammatory factors such as IL-4, IL-10 and Treg-related transcription factor Foxp3. Conclusions Our data suggest that giving 5% DSS solution to C57BL/6J mouse is appropriate to efficiently establish a murine IBD model. This laid an important foundation for further studies of the pathogenesis of IBD, biological characteristics, and intervention factors.