Based on the intestinal damage induced by deoxynivalenol (DON) in KM mice, to investigate the protective effect of apigenin (API) and reveal its potential mechanism, healthy male KM mice were randomly divided into a control group, a DON group, a low-dose API group, a high-dose API group, a low-dose API intervention group (low-dose API+DON), and a high-dose API intervention group (high-dose API+DON). The protective effects of API against DON-induced intestinal injury in KM mice were comprehensively assessed through multiple approaches: monitoring of body weight, pathological staining of colonic tissue with hematoxylin-eosin, TUNEL apoptosis staining, western blotting analysis of apoptosis-related protein expression, and measurement of serum lactate dehydrogenase (LDH), total superoxide dismutase (SOD), and malondialdehyde (MDA) levels. The results showed that, compared with the blank group, the DON group exhibited significantly decreased body weight after peaking on day 19, and high-dose API intervention successfully mitigated the trend of weight loss. Pathological examination revealed that colonic tissue damage improved after high-dose API intervention, and the pathological score of damage was reduced by 1.3 when compared with that of the DON group. TUNEL staining showed that API interventions significantly reduced the count of apoptotic cells. Western blot analysis further confirmed that API intervention down regulated the expression of apoptotic proteins Bax, P53, caspase-12, and caspase-9 and decreased the expression of Bcl-2. Serological assays revealed that, compared with the DON group, high-dose API intervention reduced LDH level by 7.53% (P<0.05), increased SOD activity by 46.82% (P<0.05), and reduced MDA level by 12.25% (P<0.05). These results indicate that API has a significant protective effect on DON-induced intestinal injury in KM mice, and the mechanism may be related to the synergistic effects of multiple pathways, including the inhibition of DON-induced apoptosis in colonic tissue as well as improvement of serum LDH and SOD activities in the mice. This study provides strong theoretical support for the potential application of API as a functional food ingredient in the prevention and treatment of DON-related intestinal injury.