In this study, rats with a fresh oil diet, a deep‐fried oil diet, and a deep‐fried oil and resistant starch (RS) diet were investigated for revealing the effects of deep‐fried oil to the metabolic system and if RS could effectively attenuate metabolic dysfunction caused by deep‐fried oil. The results showed that DO feeding led to significant increases of liver biomarkers of alanine aminotransferase (ALT) and aspartate transaminase (AST), accompanied by consistent reduction of total antioxidation (T‐AOC) and glutathione peroxidase (GSH‐Px) activity compared to the rats feeding with unheated canola oil (FO group) (p < 0.05). Liver histology of rats in DO group exhibited membrane blebbing and boundary ambiguity, indicating DO might exert significant hepatotoxic effects. However, RS intervention (DO‐RS group) significantly reversed these changes. Furthermore, the results in this study revealed that p53 and MAPK signaling pathways presented in the significantly enriched KEGG pathways list in FO versus DO group, but not in FO versus DO‐RS group, suggesting RS intervention modulated these two signaling pathways. This is the first study to investigate RS intervention on the attenuation of hepatotoxicity induced by DO intake in the dietary.