The objective of this study was to investigate the influence of ora-pro-nobis (Pereskia aculeata Mill.) flour on the adhesion of probiotics to intestinal epithelial cells and to evaluate the effect of a product based on this flour on gastrointestinal symptoms, weight, body fat, glycemia, and lipid profile in overweight men.Microbiological counts (probiotic count, survival after in vitro gastrointestinal resistance, Caco-2 cell adhesion) were analyzed. A randomized, cross-over intervention was performed. Intestinal microbiota was indirectly assessed on the basis of consistency, color of feces, and gastrointestinal symptoms.P. aculeata did not affect Lactobacillus casei adhesion to Caco-2 cells. Ora-pro-nobis flour improved gastrointestinal symptoms and increased satiety.The consumption of ora-pro-nobis flour improved intestinal health. In addition, it maintained the high adherence of L. casei to intestinal cells as well as patient anthropometric and biochemical parameters.Pereskia aculeata Mill. is well known in folk medicine and has several nutrients; however, there are few studies on this plant. This is the first study to analyze the influence of P. aculeata on bacterial adherence and the first cross-over clinical trial to evaluate the beneficial potential of ora-pro-nobis flour in overweight men. Thus, this study will contribute to the promotion of ora-pro-nobis as a functional ingredient and will arouse the interest of industries to develop related healthy foods. In addition, it is an effective dietary strategy to improve the gastrointestinal health of men.