The invention discloses a preparation method for a novel non-alcoholic fatty liver disease model. The preparation method comprises the following steps: a, randomly dividing fed mice into two groups, with one group being a model group and the other group being control mice, recording states everyday and taking urine, hepatic tissue and blood for lipid analysis; b, processing experimental subjects; c, building a model; d, collecting hepatic tissue and blood; e, preparing conventional frozen sections from hepatic tissue specimens of the mice, taking materials, wrapping the frozen sections with common glue and then successively carrying out dyeing and sealing; f, carrying out conventional pathological examination on model mice; g, carrying out conventional pathological examination on control mice; and h, comparing results of dynamic analysis of hepatic tissue fat obtained in step f and step g. The novel non-alcoholic fatty liver disease model established in the invention has the advantages of reasonable route, easy operation and strong pertinency and is an effective tool for further research on the non-alcoholic fatty liver disease.