Oats are widely recognized for their numerous health benefits, particularly regarding their anti-inflammatory properties. However, research exploring their specific effects on intestinal permeability and tight junction (TJ) integrity in the context of inflammatory bowel disease (IBD) has been limited. This study aimed to investigate the therapeutic efficacy of germinated oat extract (GOE) in managing IBD, a condition marked by persistent gastrointestinal inflammation and increasing global prevalence. The identified compounds were used to predict target biomarkers and mechanisms related to IBD via bioinformatics analysis and validated using in vitro models. In this study, we used network biology and chemical informatics approaches to predict target biomarkers and their molecular mechanisms. The predicted biomarkers were validated for their effectiveness using a cellular model of intestinal inflammation. The effectiveness of treatment with GOE was validated via in vitro studies, which demonstrated significant enhancement in transepithelial electrical resistance (TEER) and a reduction in fluorescein isothiocyanate (FITC) permeability. Analysis of the mRNA expression of IBD-associated biomarkers in Caco-2 cells demonstrated a significant increase in the mRNA levels of TJ proteins, including TJP1, TJP2, occludin, claudin-1 and claudin-3 compared to the inflammatory group. Furthermore, treatment with GOE markedly reduced the mRNA expression levels of proinflammatory cytokines such as TNF-α, IL-6, and CXCL8. The combination of COCONUT and chemical profiling analysis provided insights into the fundamental molecular mechanisms of GOE. These results underscore the potential of systematically using big data-driven network biology to analyze the effect of food components, highlighting GOE as a promising dietary intervention for IBD.