Abstract: To explore the regulatory effect of Rosa roxburghii Tratt. fruit vinegar (RFV) on nonalcoholic fatty liver disease (NAFLD) in mice, a high-fat diet (HFD)-induced NAFLD model in mice was established. The regulatory effects of RFV on NAFLD mice were investigated from various aspects, including body weight, epididymal fat mass, liver index, lipid metabolism, insulin resistance, oxidative stress, transaminase activity, and pathological sections. UPLC-MS method was developed to analyze the compounds of RFV. The results showed that, compared with the model group, the high-dose RFV had a highly significantly decreased the rate of weight gain by 36.38%, and a highly significantly decreased of epididymal fat mass by 57.03% (P<0.01), and highly significantly reductions in TG, TC, and LDL-C by 55.17%, 37.38%, 69.76% (P<0.01). Additionally, the HDL-C levels were highly significantly increased by 74.16% (P<0.01), SOD enzyme activity was highly significantly enhanced by 49.71% (P<0.01), and MDA content was highly significantly reduced by 51.96% (P<0.01). The insulin resistance index was highly significantly lowered by 45.85% (P<0.01), and the liver injury sensitivity markers ALT and AST were highly significantly down regulated by 60.03% and 53.56%, respectively (P<0.01). Therefore, it was speculated that RFV had a regulatory effect on NAFLD mice and was better than that of Rosa roxburghii Tratt. juice. Composition analysis showed that, RFV mainly contained 27 phenols compounds, 14 terpenoids, 5 organic acids, 4 sugars, 3 lactones, 3 amino acids, 2 lipids, 1 vitamin, and 11 other compounds. The number of phenols and terpenoids was up to 41. They may be the key active compounds regulating NAFLD. In conclusion, RFV was riched in various active compounds and had a significant regulatory effect on NAFLD induced by high-fat diet.