Thymol Protects against Aspergillus Fumigatus Keratitis by Inhibiting the LOX-1/IL-1ß Signaling Pathway.

ABSTRACT

OBJECTIVE: To explore the anti-inflammatory effects and mechanisms of action of thymol in Aspergillus fumigatus (A. fumigatus) keratitis. METHODS: The minimum inhibitory concentration of thymol against A. fumigatus was detected. To characterize the anti-inflammatory effects of thymol, mouse corneas and human corneal epithelial cells were pretreated with thymol or dimethyl sulfoxide (DMSO) before infection with A. fumigatus spores. Slit-lamp microscopy, immunohistochemistry, myeloperoxidase detection, quantitative real-time polymerase chain reaction, and Western blotting were used to assess infection. Neutrophil and macrophage recruitment, in addition to the secretion of LOX-1 and IL-1ß, were quantified to evaluate the relative contribution of thymol to the inflammatory response. RESULTS: We confirmed that the growth of A. fumigatus was directly inhibited by thymol. In contrast with the DMSO group, there was a lower degree of inflammation in the mouse corneas of the thymol-pretreated group. This was characterized by significantly lower clinical scores, less inflammatory cell infiltration, and lower expression of LOX-1 and IL-1ß. Similarly, in vitro experiments indicated that the production of LOX-1 and IL-1ß was significantly inhibited after thymol treatment, in contrast with the DMSO-pretreated group. CONCLUSION: Our findings demonstrate that thymol exerted a direct fungistatic activity on A. fumigatus. Furthermore, thymol played a protective role in fungal keratitis by inhibiting LOX-1/IL-1ß signaling pathway and reducing the recruitment of neutrophils and macrophages.