Aim To study the preventive effect and mechanism of ligustrazine on doxorubicin-induced cardiotoxicity in mice. Methods The mouse model of cardiotoxicity induced by doxorubicin was established. The mice were randomly divided into three groups:sham group, doxorubicin group (doxorubicin 15 mg/kg), ligustrazine treated group (ligustrazine 60 mg/(kg·d)＋doxorubicin 15 mg/kg), echocardiography was performed to detect cardiac function of doxorubicin and ligustrazine intervention groups in mice. The sections were stained with HE staining or Masson's trichrome staining for histological and collagen analysis, apoptosis of cardiomyocyte were analyzed by TUNEL staining, Western blot was used to analyze the expression of Akt, endothelial nitric oxide synthase (eNOS), the phosphorylation of Akt and eNOS.Results Compared with doxorubicin group, the fractional shortening (FS) and the ventricular ejection fraction (EF) in the ligustrazine treated group were significantly higher (P＜0.01). The ligustrazine treated group significantly decreased apoptosis of cardiomyocyte in mice as compared to the doxorubicin group (P＜0.01), the expression of p-Akt and p-eNOS in the ligustrazine treated group were stronger than those in the the doxorubicin group (P＜0.01). Meanwhile, the ligustrazine treated group significantly inhibited cleaved Caspase-3 as compared to the doxorubicin group. Conclusion Ligustrazine protects doxorubicin-induced myocardial injury by activating Akt/eNOS signaling pathway and inhibiting cardiomyocyte apoptosis, and it may be an underlying mechanism by which ligustrazine can prevent doxorubicin-induced cardiotoxicity.