This paper mainly studied the effect of Lactobacillus rhamnosus DHC32 on intestinal microflora and mucosal immune of normal mice. The results showed that DHC32 reduced the moisture， ammonia and pH of feces， and increased metabolic capacity of fecal microflora in mice. Compared to the control group， the growth of Lactobacilli and Bifidobacteria were promoted， meanwhile， the Bacteroides fragilis， Enterobacteria and other harmful facal bacteria were inhibited. The production of short chain fatty acids in feces from mice feed on DHC32 strains were significantly promoted. Moreover， DHC32 significantly increased mRNA levels of My D88， IL-23， IL-22 and other intestinal immunoregulatory cytokines. Also， the effect of DHC32 on gene expression of intestinal MUC1， MUC2 and RegIIIγ were studied. These results furtherly demonstrate that Lactobacillus rhamnosus DHC32 efficiently stabilize mucous membrane barrier， and strengthens the host immune function to improve animal intestinal health.