Type 1 diabetes (T1D) is characterized by dysregulated blood glucose and liver metabolism. In previous studies, niclosamide ethanolamine salt (NEN) and artemether (Art) displayed significant hypoglycemic effects. However, their combined therapeutic effects on the liver in T1D have remained elusive. In the present study, T1D mice were established and randomly allocated into groups. Following treatment, the physiological and metabolic parameters, including liver function, glycogen content, glucose-6-phosphatase (G6Pase) protein expression levels, mitochondrial biogenesis and mitochondrial metabolism were analyzed. Compared with the NEN or Art treatments alone, their combination improved glycometabolism and the symptoms of diabetes. Combined treatment with NEN and Art also significantly ameliorated liver injury and increased liver glycogen storage. Furthermore, combinatorial treatment significantly downregulated hepatic G6Pase protein expression levels and regulated mitochondrial biogenesis. NEN and Art increased the respiratory exchange rate and reduced mitochondrial phosphoenolpyruvate carboxykinase and branched-chain α-keto acid dehydrogenase complex protein expression levels, whereby the effects were obviously enhanced by their application as a combined treatment. In conclusion, the present study confirmed that combined treatment with NEN and Art improved glycometabolism and liver function in T1D mice and the therapeutic effects may be partially associated with the regulation of liver mitochondria.Copyright: © Weng et al.