Background: To investigate the protective effect of Ixeris Sonchifolia (Bae.) Hance (ISH) extract on herpes simplex virus keratitis (HSK) in mice. Methods: A mouse model of HSK was established by inoculating 60 mice (60 right eyes) with herpes simplex virus type 1 (HSV-1) by corneal scratch. The other 15 mice as blank control only received corneal scratch but without HSV-1. From the 2nd day after the successful modeling, the experimental group was fed with ISH total flavonoids (50, 100 and 200 mg/kg) orally, twice a day for 14 days. The model group and control group were given the same amount of normal saline. The pathological changes of cornea were observed once a day by slit lamp microscopy combined with fluorescein staining. The corneal histopathological examination, the survival status and the serum levels of interleukin-2 (IL-2), IL-4 and interferon-gama (INF-γ) were performed at the end of the experiment. Results: The result showed that ISH could significantly improve the corneal lesion degree, increase mice survival rate, and markedly increase the levels of IL-2 and INF-γ, reduce the levels of IL-4 in serum of mice. Conclusions: ISH could increase the anti-virus ability, promote the healing of corneal inflammation and alleviate the pathological damage of cornea, which suggested that ISH has a potential and valuable therapeutic effect on the HSK.