Doxorubicin (DOX) for treatment of patients with neoplasms shows cardiac toxicity. Recent studies reported that histamine H2 receptor antagonist improved DOX-induced cardiotoxicity, while histamine deficiency exacerbates myocardial injury. To elucidate the effects and mechanisms of histamine on DOX-induced acute cardiotoxity, we investigated the cardiac functions, pathological alterations, and protein expressions of Hsp25, LAMP-1, Beclin-1, p62, LC3B, histamine H1 and H2 receptors using both mice with organic cation transporter-3 (OCT3) deficiency and mice with supplementation of L-histidine that is converted to histamine via histidine decarboxylase. The DOX-induced acute cardiotoxicity was improved in both mice by analyzing cardiac function with echocardiography and electron microscopy. The higher content of histamine, decreased histamine H₂ receptor expression in basal levels, improvements of autophagy-lysosome flow, and enhancement of Hsp25 protein expression were shown in OCT3(-/-) deficiency and mice with supplementation of L-histidine. In conclusion, OCT3 deficiency and supplementation of L-histidine significantly contributes to DOX-induced acute cardiotoxicity.