Hydrogen sulfide (H₂S) is a toxic gas getting recognized as an endogenous signaling molecule in multiple organ systems, in particular cardiovascular system. H₂S is known to regulate cardiac function and protect against ischemic injury. However, little information is available regarding the effect of H₂S on cardiac function in insulin resistance. This study was designed to examine the impact of H₂S supplementation on cardiac function using an Akt2 knockout model of insulin resistance. Wild-type and Akt2 knockout mice were treated with NaHS (50 micromole/kg/d, i.p. for 10 days) prior to evaluation of echocardiographic, cardiomyocyte contractile and intracellular Ca²⁺ properties, apoptosis and mitochondrial damage. Our results revealed that Akt2 ablation led to overtly enlarged ventricular end systolic diameter, reduced myocardial and cardiomyocyte contractile function, disrupted intracellular Ca²⁺ homeostasis and apoptosis, the effects of which were ameliorated by H₂S. Furthermore, Akt2 knockout displayed upregulated apoptotic protein markers (Bax, Caspase-3, -9 and -12) and mitochondrial damage (reduced aconitase activity and NAD⁺, elevated cytochrome C release from mitochondria) along with reduced phosphorylation of PTEN, Akt and GSK3β in the absence of changes in pan protein expression, the effects of which were abolished or significantly ameliorated by H₂S treatment. In vitro data revealed that H₂S-induced beneficial effect against Akt2 ablation was obliterated by mitochondrial uncoupling. Taken together, our findings suggest the H₂S may reconcile Akt2 knockout-induced myocardial contractile defect and intracellular Ca²⁺ mishandling, possibly via attenuation of mitochondrial injury and apoptosis.