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Our experiments were designed to study the effect of diltiazem (DIL) combined with superoxide dismutase (SOD) on myocardial ischemia-reperfusion (MIRI) injury in a rat model.Fifty rats were randomly separated into sham, ischemia-reperfusion (IR), DIL (5 mg/kg), SOD (10,000 U/kg) and combinatorial therapy (DIL plus SOD) groups. MIRI was induced by ligating the left anterior descending coronary artery for 30 min and then reperfusing for 60 min. The cardioprotective effects of combinatorial therapy were evaluated using hemodynamics, biochemical indices, histopathology and apoptotic-related proteins and gene expression.Compared with the IR group, combinatorial therapy significantly improved cardiac function and decreased arrhythmia, myocardial infarction area and release of myocardial enzyme. In addition, combinatorial therapy protected the myocardial cell structure as well as markedly alleviated oxidative stress, resulting in upregulation of Bcl-2 and adenine nucleotide transporter-1 expression as well as downregulation of Bax, caspase-3 and cleaved caspase-3 expression.Our results indicated that DIL combined with SOD can provide protection against MIRI in rats, and these effects may be attributed to a reduction in oxygen stress damage, attenuation of calcium overload, and inhibition of cell apoptosis.