Objective: The aim of this study was to investigate the protective effect of eugenol against Fusarium solani (F. solani)-induced fungal keratitis (FK) in mice and to preliminarily explore possible underlying mechanisms. Methods: FK mouse models were prepared using a modified epifluorescence microscopy method. All mice were coated with equal amount of DMSO (0.05%) in the left eye as a blank control. Equal amount of DMSO (0.05%) was applied to the conjunctiva of the right eye of rats in the model group. The eugenol group was treated with 4 μL of eugenol (160 μg/mL) applied to the conjunctival sac of the right eyes of the mice. The insulin-like growth factor-1 (IGF-1) group was coated with the PI3K/AKT pathway activator IGF-1 (10 nmol) in the conjunctival sac of the right eye in addition to the administration of eugenol. The corneal morphology was observed under a slit lamp microscope on days 1, 3, and 5 of inoculation with F. solani suspension, respectively. Hematoxylin eosin (HE) staining was used to assess corneal histopathological damage. Corneal tissue bacterial load was determined. Enzyme-linked immunosorbent assay and protein immunoblotting were used to analyze the levels of inflammatory mediators interleukin-6 (IL-6) and interleukin-1β (IL-1β) and the expression of PI3K/AKT pathway proteins. Results: Eugenol treatment improved morphological symptoms of keratitis and inflammatory response in FK mice, and reduced corneal pathologic tissue damage and fungal load. In the early stage of F. solani infection, corneal tissue IL-6 levels were significantly higher and IL-1β levels were significantly lower in the eugenol group compared with the model group (P<0.05); and corneal tissue IL-6 levels in the eugenol group were significantly higher than, and IL-1β levels were significantly lower than, those in the IGF-1 group (P<0.05). In the late stage of infection, both IL-6 and IL-1β levels in corneal tissues of the eugenol group were significantly lower than those of the model and IGF-1 groups (P<0.05). Compared with the model group, the expression of both p-PI3K and p-Akt in corneal tissues of the eugenol group was significantly lower (P<0.05); and the expression of p-PI3K and p-Akt in corneal tissues of the eugenol group was significantly lower than that of the IGF-1 group (P<0.05). Conclusion: Eugenol may attenuate F. solani induced corneal inflammation by inhibiting the PI3K/AKT pathway, and has a protective effect against F. solani keratitis in mice.