Multi-omics analysis and experimental verification reveal the role of dihydrocaffeic acid against 5-fluorouracil-induced intestinal mucositis

BACKGROUND: A common adverse effect of 5-fluorouracil (5-FU) in chemotherapy is intestinal mucositis (IM), which significantly impacts patients' quality of life. To date, no effective long-term treatment is available for IM. Dihydrocaffeic acid (DHCA), a bioactive dietary compound from Gynura bicolor, exhibits potent anti-inflammatory and antioxidant properties. However, the exact mechanisms through which DHCA mediates its preventive and therapeutic effects on 5-FU-induced IM remain unclear. PURPOSE: This study aims to evaluate the protective and therapeutic potential of DHCA against 5-FU-induced intestinal mucositis and to elucidate the underlying molecular mechanisms. METHODS: An intestinal mucositis model was successfully established using 5-FU to explore the effects of DHCA. Subsequently, multi-omics and network pharmacology analyses were performed to elucidate the pathways potentially implicated in the effects of DHCA. Validation experiments were conducted in both mouse and cell models. RESULTS: In vivo experiments demonstrated that prophylactic administration of DHCA markedly attenuated multiple 5-FU-induced adverse effects, including body weight loss, elevated colonic inflammation scores, excessive production of pro-inflammatory cytokines, goblet cell depletion, colon shortening, and disruption of tight junction integrity. Integrated multi-omics and network pharmacology analyses further suggested that the protective effects of DHCA are closely associated with the regulation of inflammation- and lipid metabolism-related signaling pathways. Molecular docking and results from multiple experiments revealed a direct interaction between DHCA and p38 MAPK, supporting its role in suppressing inflammatory signaling. In addition, DHCA enhanced antioxidant defense through activation of the Nrf2/HO-1/GPX4 axis, as evidenced by reduced MDA levels and increased SOD activity and GSH content, thereby alleviating oxidative stress. These in vivo-identified protective effects and underlying mechanisms were further confirmed by in vitro experiments. CONCLUSION: Our findings demonstrate that DHCA shows inflammation-suppressing, ROS-scavenging, and apoptosis-inhibiting properties in 5-FU-induced IM. As a natural compound, DHCA holds promise as a novel drug for the treatment and prevention of 5-FU-induced IM.