Abstract
Purpose: This study aimed to investigate how Fusarium solani triggers PANoptosis in fungal keratitis and to examine the toxic impacts and underlying mechanisms of deoxynivalenol (DON) on human corneal epithelial (HCE-T) cells. Methods: A F. solani keratitis mouse model was established through intrastromal injection of fungal spores. Clinical severity was evaluated using a standardized scoring system. PANoptosis markers in corneal tissues were analyzed via western blotting, immunofluorescence, and TUNEL assay. Real-time PCR was used to quantify inflammatory cytokines. RNA sequencing was conducted to select differentially expressed genes (DEGs) and analyze immune cell infiltration profiles. HCE-T cells were exposed to DON, and PANoptosis-related proteins were detected via western blot and immunofluorescence. Gene expression alterations were analyzed using RNA sequencing. Results: PANoptosis activation was validated through upregulated expression of cleaved caspases-1, -3, -7, and -8; gasdermin D (GSDMD); and phosphorylated mixed lineage kinase domain-like protein (p-MLKL) in infected corneas, accompanied by augmented TUNEL-positive cells. Inflammatory cytokines (IL-6, IL-1β, TNF-α) were significantly upregulated. RNA sequencing revealed significant changes in PANoptosis, immune response, and extracellular matrix organization. Immune infiltration profiling indicated a marked increase in M1 macrophages in infected corneas. In vitro experiments demonstrated that DON exposure increased PANoptosis markers (cleaved caspases-1, -3, -7, and -8; BAX; GSDMD; and p-MLKL) in HCE-T cells, along with upregulation of inflammatory cytokines (IL-6, IL-1β, IL-18, TNF-α). RNA sequencing further revealed alterations in ribosomal RNA (rRNA) processing and extracellular matrix organization pathways in DON-treated cells. Conclusions: F. solani infection and DON exposure induce PANoptosis in fungal keratitis, leading to significant inflammatory responses. These results suggest that targeting PANoptosis signaling pathways may represent a novel therapeutic approach for treating fungal keratitis.