Non-alcoholic fatty liver disease (NAFLD), also referred to metabolic associated fatty liver disease (MAFLD), displays characteristics indicative of metabolic disorders. Despite extensive research efforts, the underlying pathogenesis of NAFLD remain largely unknown and currently lack recommended treatment options. It is widely acknowledged that obesity and type 2 diabetes are significant risk factors for NAFLD, with the accumulation of free fatty acids (FFA) in the liver considered an initial trigger for its development. In this study, we exposed the human liver cell line Huh7 to a high concentration of a mixture containing both saturated (palmitic acid) and unsaturated (oleic acid) fatty acids for 24 hours. TMT and LC-MS based proteomics yielded a set of 172 significantly regulated proteins.