Inflammatory bowel disease is a chronic disease of the intestinal tract, which is related to increased levels of various inflammatory mediators. This study aims to explore the anti-inflammatory mechanism of small molecular peptide WLS and its alleviating effect on inflammatory bowel disease (IBD). In TNF-α-induced HT-29 cells, WLS inhibited IL-8 secretion, decreased gene expression of pro-inflammatory cytokines IL-8, IL-6, IL-1β, and TNF-α, and inhibited the activation of MAPK/NF-κB signaling pathways. In the dextran sulfate sodium salt (DSS) induced colitis mouse model, WLS inhibited weight loss and disease activity index scores, increased colon length, improved colon histopathology, inhibited secretion of IL-6 and TNF-α in the colon, and down-regulated gene expression of pro-inflammatory cytokines (IL-6, TNF-α, IL-1β, IFN-γ, IL-17A). This study revealed that WLS was a novel small molecule peptide with anti-inflammatory activity and may be a potential candidate for the treatment of inflammatory bowel disease.