Viral myocarditis (VMC) is closely related to apoptosis, oxidative stress, innate immunity, and energy metabolism, which are all linked to mitochondrial dysfunction. A close nexus between mitochondrial dynamics and cardiovascular disease with mitochondrial dysfunction has been deeply researched, but there is still no relevant report in viral myocarditis. In this study, we aimed to explore the role of Dynamin-related protein 1 (Drp1)-linked mitochondrial fission in VMC. Mice were inoculated with the Coxsackievirus B3 (CVB3) and treated with mdivi1 (a Drp1 inhibitor). Protein expression of Drp1 was, increased in mitochondria while decreased in cytoplasm and accompanied by excessive mitochondrial fission in VMC mice. In addition, midivi1 treatment attenuate inflammatory cells infiltration in myocardium of the mice, serum Cardiac troponin I (C-TnI) and Creatine kinase-MB (CK-MB) level. Mdivil also could improved the survival rate of mice and mitochondrial dysfunction reflected as the up regulated mitochondrial marker enzymatic activities of succinate dehydrogenase (SDH), cytochrome c oxidase (COX) and mitochondrial membrane potential (MMP). At the same time, mdivil rescued the body weight loss, myocardial injury and apoptosis of cardiomyocyte. Furthermore, decease in LVED5 and increase in EF and FS were detected by echocardiogram, which indicated the improved myocardial function. Thus, Drpl-linked excessive mitochondrial fission contributed to VMC and midivil may be a potential therapeutic approach. (C) 2017 Elsevier Inc. All rights reserved.