The invention relates to a prevention effect of rape pollen total flavonoids on non-alcoholic fatty liver induced by high fat. The method comprises the following steps: feeding a mouse with high fat for 10 weeks, establishing a non-alcoholic fatty liver mouse model, carrying out intragastric administration intervention while molding, carrying out intragastric administration intervention once a day, after 10 weeks, measuring AST and ALT levels in mouse serum, TG, TC, LDL-C and HDL-C contents and TG and TC content changes in liver tissues, and observing morphological changes of the liver tissues by an HE staining method. Compared with a model group, the rape pollen total flavonoids can significantly reduce the body weight of mice, reduce the levels of AST and ALT in serum, reduce the contents of TG, TC and LDL-C in serum, reduce the contents of TC and TG in liver tissues, increase the content of HDL-C in serum, and significantly improve the pathological morphology of the liver tissues of the mice. The rape pollen total flavonoids can prevent the non-alcoholic fatty liver induced by high fat by means of improving liver functions, regulating blood fat level, improving liver tissue lesions and the like.