Abstract: The aim of this study was to investigate the protective effects of exogenous 5'-nucleotides on alcohol induced colon function injury in SD rats.Fifty SPF male Wistar rats were randomly divided into normal control group, alcohol model group, dextrose control group, 0.04% and 0.16% nucleotides intervention group. After continuous ethanol lavage for 6 weeks, the results showed that exogenous 5'-nucleotides could effectively improve the pathological changes of colon in rats induced by alcohol, increase the expression levels of Claudin and Occludin and reduce the levels of TLR4 and CD14 in colon tissues ( p <0.01) .The numbers of Bifidobacterium ( 8.32 vs.8.00 log CFU/g) and Lactobacillus ( 8.13 vs.7.44 log CFU/g) in ileum contents were significantly decreased, and the numbers of Enterococcus ( 7.03 vs.7.52 log CFU/g) and E. coli ( 6.19 vs.6.85 log CFU/g) were significantly decreased in nucleotides intervention groups when compared with the alcohol model group ( p < 0.05) .Therefore, exogenous 5'-nucleotides could improve the alcohol induced colon function injury in rats which may attribute to the regulation of gut microbiota.