AbstractInflammatory bowel disease (IBD), a common chronic gastrointestinal disease in humans, has emerged as a global public health challenge. Dihydromyricetin (DHM) has anti‐inflammatory and antioxidant activities, which can alleviate inflammation. In this study, we explored the effect and underlying mechanism of DHM on dextran sulfate sodium (DSS)‐induced colitis in mice and porcine jejunum epithelial cells (IPEC‐J2) exposed to lipopolysaccharide (LPS). We found that DHM alleviated loss of weight, diarrhea, and damage of colon structure in colitis mice. For the intestinal microbial, a significant rise in the amount of the potentially beneficial genera and a decline in the amount of harmful genera were observed in DHM‐treated colitis mice. Metabolomic analysis of cecal content revealed that DHM restored phenylalanine metabolism, arginine biosynthesis, and arachidonic acid metabolism disorders caused by intestinal inflammation. Moreover, DHM decreased the level of pro‐inflammatory cytokines and reactive oxygen species in LPS‐treated IPEC‐J2 cells. DHM also reduced the expression of MyD88 and nuclear factor‐κB (NF‐κB). In summary, we found that 125 mg/kg DHM administration alleviated diarrhea, reinstated intestinal barrier function, modulated intestinal dysbiosis, and suppressed the expression of myeloid differentiation factor 88 (MyD88) and NF‐κB. Therefore, DHM may be a potentially therapeutic agent for IBD.