Study On The Mechanism Of Rapamycin On Fungal Keratitis In Mice Infected By Aspergillus Fumigatus

Purpose: To observe the influence of rapamycin on the cornea infected by Aspergillus fumigatus in mice.To investigate the effect of rapamycin on regulating the responses of innate immunity in A.fumigatus keratitis.To explore the pathway of rapamycin against inflammation in fungal keratitis.Methods: The prepared healthy 6-8 weeks old female C57BL/6 mice were randomly divided into rapamycin solvent control group(group A)and rapamycin drug group(group B).Among them,A and B groups were divided into normal cornea group and fungal keratitis group infected with Aspergillus fumigatus.Photographs of mouse corneal infections were taken at 0,12 h,1 day and 3 days after infection to observe the corneal infections,and clinical scores and statistics were performed.The mice were sacrificed and the corneas of the mice were harvested.The expression of m TOR(mammalian rapamycin target),LOX-1(lectin-like oxidized low-density lipoprotein receptor 1),and proinflammatory cytokines IL-1?,IL-6,MIP-2,and IFN-? were detected by real time-PCR(6 eyes/group/time);Protein expressions of p-m TOR(phosphorylated m TOR),LOX-1,IL-1?,and IL-6 were detected by Western Blot.The immunofluorescence technique was used to verify the change of expression of LOX-1.Results: Rapamycin treatment decreased the clinical scores and production of proinflammatory cytokines IL-1?,IL-6,MIP-2 and IFN-? in C57BL/6 cornea after infection.The protein expression of p-m TOR was down-regulated by rapamycin.Expression levels of LOX-1 stimulated by A.fumigatus were also reduced by rapamycin.Conclusion: Compared to the solvent control group,immunosuppressant rapamycin significantly reduced the clinical score of Aspergillus fumigatus-infected fungal keratitis and plays an anti-inflammatory role in innate immunity of fungal keratitis in mice.The anti-inflammatory effect could be through m TOR/LOX-1 signaling pathway during A.fumigatus infection.