Diabetic cardiomyopathy (DCM) describes pathological changes in the myocardium in diabetes and is not directly attributable to coronary artery disease or hypertension. The cardioprotective effect of rosemary extract and hesperidin against Streptozotocin-induced diabetic cardiomyopathy in rats was evaluated. Fifty rats were divided into five groups. Group Ι: rats fed a normal diet. Group Π: rats received a single intraperitoneal injection of streptozotocin (50 mg /kg body wt.). Group III: rats treated with insulin (2 U/rat per day). Group IV: rats administered orally rosemary extract at a dose of 200 mg/kg body weight/day. Group V: rats were given hesperidin orally at a dose of 100 mg/kg body weight/day. Blood samples and heart tissue specimens were collected after 12 weeks of the experiment for determination of some biochemical and molecular biomarkers. The obtained results showed a marked increase in blood glucose concentration, creatine kinase-MB and lactate dehydrogenase activities in addition to a significant upregulation of Histone acetyltransferase (HAT) and Histone deacetylase (HDAC) with downregulation of Glucose transporter 4 (GLUT4) gene expression in cardiac tissue of diabetic cardiomyopathy. Treatment with insulin, rosemary extract, or hesperidin exhibited a significant decrease in blood glucose level, CK-MB, and LDH activities in addition to a significant down-regulation of HAT and HDAC. However, GULT4 translocation showed a significant upregulation. It was concluded that rosemary extract and hesperidin have potential therapeutic in DCM by attenuating GLUT4 Pathway involved in the glucose uptake by the myocardium in diabetic rats.