Aims: This study aimed to evaluate the effects and mechanisms of tertiary butylhydroquinone (TBHQ) on insulin resistance (IR) and diabetic liver steatosis. Methods: Male ApoE-/- mice were received streptozocin (STZ) injection and a high-sugar-high-fat diet to form type 2 diabetes mellitus (T2DM). Then, the mice were given TBHQ for six weeks. Body weight, fasting blood-glucose (FBG), postprandial blood glucose (PBG), insulin and oral glucose tolerance test (OGTT) were detected on all the mice. Hematoxylin-eosin staining and western-blot were performed to detect the morphological structure and the target proteins expression in liver tissues. In vitro, HepG2 cells were induced by HClO and insulin to develop IR. Western-blot was used to evaluate the related proteins expression. Hoechst staining was conducted to measure cell apoptosis. Results: Mice that received STZ injection and a high-sugar-high-fat diet developed T2DM. TBHQ reduced blood glucose level, improved glucose tolerance, alleviated liver steatosis in diabetic mice. Moreover, TBHQ significantly increased AMPKα2, GLUT4 and GSK3β expression, up-regulated PI3K and AKT phosphorylation level in diabetic mice liver. Notably, TBHQ down-regulated HClO and insulin-induced cell IR and inhibited cell apoptosis via AMPKα2/PI3K/AKT pathway. Conclusion: TBHQ alleviated IR and liver steatosis in T2DM mice and the mechanism may relate to AMPKα2/PI3K/AKT pathway.