In this study, we assessed the effects of Radix Rehmanniae Preparata polysaccharides (RRPP) on the digestive enzymes, morphology, microbial communities and mucosal barrier function of the intestine of Luciobarbus capito. Fish were fed different amounts of RRPP (0, 0.05, 0.1, 0.2 and 0.4%) for 60 days. The results revealed that intestinal amylase and trypsin activities significantly increased after the 0.2% RRPP treatment (p < .05). Increases in intestinal fold height, wall thickness and fold density were observed in Luciobarbus capito fed an RRPP‐supplemented diet, and the best effect was observed in the proximal intestine. Sequencing of the bacterial 16S rRNA V4 region showed that dietary RRPP reduced intestinal microbial community richness and species diversity. Fusobacteria, Proteobacteria, Firmicutes and Verrucomicrobia were the dominant bacterial phyla in the intestine of Luciobarbus capito. Dietary RRPP affected the overall microbial community and increased the abundance of the genus Cetobacterium. Moreover, RRPP significantly upregulated the expression of occludin, zonula occludens (ZO)‐1 and claudin‐c in the intestine (p < .05). In summary, supplementing the diet of Luciobarbus capito with RRPP can improve digestive enzymes activities and intestinal morphology, modulate the microbial composition and enhance intestinal mucosal barrier function.