Herpes simplex virus type 1 (HSV-1) had led to kinds of clinical disorders and became an important cause of morbidity and mortality worldwide, such as herpetic gingivostomatitis in children. Previous studies reported that HSV-1 infection is common and has evolved a variety of mechanisms to evade the immune system, such as dysregulation of miRNAs. However, reports concerning the role of miRNA in HSV-1infection are limited. Here, we report that a host microRNA, miR-373, was significantly upregulated by HSV-1 infection in Hela cells and patients with herpetic gingivostomatitis and it facilitated HSV-1 replication in vitro. Subsequently, we demonstrated that miR-373 was a negative regulator of IFN-I response by directly targeting IRF1, resulting in the suppression of interferon stimulated gene (ISG) expression and enhancement of HSV-1 infection. Taken together, our findings provide new evidence of which HSV-1 hijacks the host miRNAs to promote its replication by negatively regulating the production of type I IFN and suggest a novel potential anti-HSV-1 therapeutic target.