This study aimed to investigate the effects of PNZ on intestinal mucosal barrier function in weaning piglets. A total of 210, 21-day-old piglets with similar body weights (6.30 ± 0.51 kg) were randomly allocated into seven groups: control group (CON), antibiotic group (ANT), ZnO group (ZO), nano-ZnO group (NZO) and low, middle, and high PNZ groups (LPNZ, MPNZ, and HPNZ). The seven groups were, respectively, fed control diets or control diets supplemented with antibiotics; 3,000 mg/kg ZnO; 800 mg/kg nano-ZnO; 700, 1,000, or 1,300 mg/kg PNZ. More integrated intestinal villi were observed in the LPNZ group. In the jejunum of LPNZ group, the crypt depth significantly decreased (P < 0.05), and the ratio of villus height to crypt depth (V/C) significantly increased (P < 0.05). In addition, the villus width and surface area of the ileum were significantly increased in the LPNZ group (P < 0.05). Dietary supplementation with PNZ can significantly increase the number of goblet cells in the mucosa of the jejunum and ileum (P < 0.05), decrease the contents of TNF-α and IL-1β (P < 0.05), and increase the contents of sIgA and IL-4 in the jejunal and ileal mucosa (P < 0.05). Meanwhile, the mRNA expression of MCU2 and ZO1 in PNZ group were significantly increased (P < 0.05), the mRNA expression of TLR4 and MyD88 was downregulated (P < 0.05). With increasing levels of PNZ, decreased proinflammatory cytokines and increased intestinal mucosal barrier function in weaned pigs was observed. In conclusion, supplementation with PNZ could effectively improve the intestinal barrier function of weanling piglets and potentially could replace the use of high doses of ZnO and antibiotics. The appropriate dose of PNZ for supplementation was 700 mg/kg.