Intestinal barrier dysfunction plays a vital role in the pathogenesis of inflammatory bowel disease (IBD) and may contribute to the pathology of obesity and other diseases. The goal of this study was to investigate the effects of aqueous extract of passion fruit (Passiflora edulis) leaves on intestinal barrier dysfunction induced by lipopolysaccharides (LPS) from Escherichia coli and interleukin-1β (IL-1β). Caco-2 cells were differentiated on transwell inserts and used as a model of intestinal barrier permeability. Transepithelial electrical resistance (TEER) was used as indicator of monolayer integrity. Differentiated cells were treated with LPS (10 μg.mL−1) and IL-1β (10 ng.mL−1 ) as inflammatory stimulus (IS) and passion fruit leaves extract (PFLE) at 5 e 10 mg.mL−1. LPS/IL-1β-stimulated Caco-2 cells were incubated for three hours and received in the apical side the treatment with PFLE for more three hours. These treatments were compared to the control treatment (C), consisting in only culture medium (DMEM) containing 1% fetal bovine serum (FBS). After 24 hours, the C treatment kept TEER values at 96.73%, compared to 100% initial TEER measures, showing the maintenance of the monolayer integrity. The IS reduced TEER by 40.22%, which confirm the activity of the IS on decreasing monolayer integrity. After PFLE treatment, the TEER decrease was attenuated, (12.84%, p < 0.05), compared to IS and C. Our results reveal a protective effect of the extract associated with the improvement of intestinal integrity. PFLE can be considered a candidate as a dietary supplement for intestinal disorders and to promote barrier function.