Diabetic cardiomyopathy (DCM) is a myocardial disorder resulting from glucose metabolism dysfunction, leading to structural and functional heart abnormalities independent of common cardiovascular conditions. This study explores the impact of Mammalian Sterile20-like Kinase 1 (Mst1) and Nuclear Factor E2-Related Factor 2 (Nrf2) pathways on autophagy in type 2 diabetic mice. By employing Mst1 knockout and Nrf2 activation, improvements in cardiac function reduced myocardial fibrosis, and decreased cardiomyocyte apoptosis was observed, with enhanced autophagy noted in Mst1 knockout mice further augmented by Nrf2 activation. The Mst1/Nrf2 pathway demonstrates a protective effect by regulating autophagy-related proteins, offering a potential therapeutic avenue for treating DCM in type 2 diabetes.