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Nonalcoholic fatty liver disease (NAFLD) is a major global health problem with few therapeutic options available so far. Accumulating evidence suggests that probiotics have beneficial effects on NAFLD by modulating gut microbiota. Bifidobacterium lactisProbio‐M8 (M8) is a new probiotic strain isolated from human breast milk. The aim of this study was to investigate whether M8 could protect against NAFLD in rats fed a high‐fat diet by modulating gut microbiota. In this study, rats were randomly distributed into four groups: normal diet (ND) group, normal diet plus M8 (ND+M8) group, high‐fat diet (HFD) group, and high‐fat diet plus M8 (HFD+M8) group. Ten weeks later, hepatic morphological changes and biochemical indicators were measured. 16S rDNA sequencing was applied to analyze the gut microbiota alterations. Our results showed that M8 administration effectively improved hepatic steatosis and liver damage in high‐fat diet‐fed rats. 16S rDNA analysis of gut microbiota indicated that M8 could modulate the gut microbiota composition, especially increasing Bifidobacteriumand decreasing Bilophila, Lachnoclostridium, GCA‐900066225, and Phascolarctobacteriumin high‐fat diet‐fed rats. In conclusion, our findings demonstrated that M8 could protect against NAFLD in rats fed a high‐fat diet, which may be attributed to the modulation of gut microbiota. Bifidobacterium lactisProbio‐M8 improved hepatic steatosis and liver damage. B. lactisProbio‐M8 modulated the gut microbiota composition in rats. This study provides new insights into the application of B. lactisProbio‐M8 to prevent nonalcoholic fatty liver disease (NAFLD).