Aim: To explore the underlying influence of HSV type-1 (HSV-1) infection on the energy metabolism of human umbilical cord-derived mesenchymal stem cells (UCMSCs). Methods: UCMSCs (derived from different donors) were isolated from umbilical cord tissue, cultured and infected with HSV-1. Various virology and biochemical assays were used to assess cell viability and function, such as plaque formation assay and mitochondrial mass assay. Results: HSV-1 infection sharply activated mitochondrial biogenesis, increased glucose consumption, oxidative phosphorylation and glycolysis of UCMSCs. Treatment with rotenone (a metabolism antagonist) and iodoacetic acid significantly blocked the proliferation of HSV-1 in UCMSCs. Conclusion: This study demonstrates, for the first time, that HSV-1 infection affects the energy metabolism process of UCMSCs. Treatment with the appropriate metabolism antagonists might improve the safety and efficacy of clinical stem cell therapies.