This study investigates the therapeutic potential of ebselen, an organoselenium compound, in treating fungal keratitis. Ebselen has been shown to have antifungal, antibacterial, anti-inflammatory, and oxidative stress-regulatory properties. The research demonstrates that ebselen effectively inhibits *Aspergillus fumigatus* growth, reduces mycelial growth, and disrupts fungal biofilm formation. The antifungal activity of ebselen is attributed to its ability to damage the fungal cell membrane through oxidative stress caused by inhibition of the thioredoxin (Trx) system. In contrast, ebselen enhances the antioxidation of the Trx system in mammalian cells. Ebselen also suppresses inflammatory mediators (IL-1β, IL-6, TNF-α, COX-2, iNOS, and CCL2) and reduces oxidative stress indicators (ROS, NO, and MDA) in fungi-stimulated RAW264.7 cells. Additionally, ebselen regulates PI3K/Akt/Nrf2 and p38 MAPK signaling pathways, contributing to improved inflammation and oxidative stress. In a mouse model of fungal keratitis, ebselen improved prognosis, reduced fungal burden, and decreased inflammatory mediator expressions and macrophage and neutrophil infiltration in the cornea. These findings suggest that ebselen is a promising treatment for fungal keratitis by reducing fungal load and protecting host tissues.This study investigates the therapeutic potential of ebselen, an organoselenium compound, in treating fungal keratitis. Ebselen has been shown to have antifungal, antibacterial, anti-inflammatory, and oxidative stress-regulatory properties. The research demonstrates that ebselen effectively inhibits *Aspergillus fumigatus* growth, reduces mycelial growth, and disrupts fungal biofilm formation. The antifungal activity of ebselen is attributed to its ability to damage the fungal cell membrane through oxidative stress caused by inhibition of the thioredoxin (Trx) system. In contrast, ebselen enhances the antioxidation of the Trx system in mammalian cells. Ebselen also suppresses inflammatory mediators (IL-1β, IL-6, TNF-α, COX-2, iNOS, and CCL2) and reduces oxidative stress indicators (ROS, NO, and MDA) in fungi-stimulated RAW264.7 cells. Additionally, ebselen regulates PI3K/Akt/Nrf2 and p38 MAPK signaling pathways, contributing to improved inflammation and oxidative stress. In a mouse model of fungal keratitis, ebselen improved prognosis, reduced fungal burden, and decreased inflammatory mediator expressions and macrophage and neutrophil infiltration in the cornea. These findings suggest that ebselen is a promising treatment for fungal keratitis by reducing fungal load and protecting host tissues.