Objective To observe changes in voltage-dependent anion channel 3(VDAC3) in a mouse model of sepsis-induced myocardial injury and to explore its potential mechanism. Methods Twenty male C57BL / 6J mice were divided randomly into a Sham group and Sepsis group, respectively (n = 10 mice per group). Sepsis was induced by the cecal ligation and puncture (CLP). Serum levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α, creatine kinase MB (CK-MB), and cardiac troponin T ( cTnT) were detected by enzyme-linked immunosorbent assay. Pathological changes in heart tissue were observed by hematoxylin and eosin staining. Structural and functional changes in the heart were evaluated by echocardiography. Changes in total glutathione, reduced glutathione (GSH),oxidized glutathione, and malondialdehyde ( MDA) in heart tissue were detected by spectrophotometry. The morphological structure of mitochondria in mouse cardiomyocytes was observed by transmission electron microscopy.Expression levels of IL-6, IL-1β, VDAC3, glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11), lipocalin-2 ( LCN2), and prostaglandin-endoperoxide synthase 2 ( PTGS2) mRNA were detected by real-time quantitative polymerase chain reaction and the localization and expression of VDAC3 and GPX4 proteins in mouse heart tissue were detected by immunofluorescence staining. The correlations between VDAC3 mRNA and GPX4, SLC7A11, PTGS2, LCN2, IL-6, and IL-1β mRNA were analyzed. Expression levels of VDAC3, GPX4, and SLC7A11 proteins were detected by Western blot. Results IL-6, TNF-α, CK-MB, and cTnT levels were significantly higher in the Sepsis group compared with the Sham group (P<0. 05). In the Sepsis group, myocardial fibers were torn, the ventricular wall was thickened and edematous, the mitochondrial membrane was ruptured, and mitochondrial cristae were broken or absent. GSH levels were significantly reduced in the Sepsis group (P<0. 05) and the lipid peroxide MDA was increased in the Sepsis group (P<0. 05) compared with the Sham group. VDAC3, GPX4 and SLC7A11 mRNA and protein levels were all lower in the Sepsis group compared with the Sham group (P<0. 05),while expression levels of IL-6, IL-1β, LCN2, and PTGS2 mRNA were increased (P<0. 05). VDAC3 mRNA was positively correlated with GPX4 and SLC7A11 mRNA levels, and negatively correlated with LCN2, PTGS2, IL-6,and IL-1β. Conclusions VDAC3 expression decreases in myocardial injury, and it may participate in the occurrence of sepsis-induced myocardial injury by regulating ferroptosis.