The Function And Mechanism Of Specialized Proresovling Meditor Maresin1 In Aspergillus Fumigatus Keratitis

Purpose: To investigate the function and mechanism of specialized proresolving meditors Maresin1 in Aspergillus fumigatus(A.fumigatus)keratitis.Methods: 1.Murine model of A.fumigatus corneal infection at 1d,3d and 5d was established by intrastromal injection.Two hours after infection,maresin1(5 ng/5 ?l)was delivered by subconjunctival injection.Then,topical administration of maresin1(5 ng/3 ?l)was applied to mouse corneas twice a day from day 1 to day 5.Control group was received same volume PBS by the same way.The development of FK lesions,the production of chemokines,the production of inflammation cytokines and the levels of p-GSK3? were measured via slit-lamp biomicroscope,quantitative polymerase chain reaction(q RT-PCR)and western blot.The presence of neutrophils in the cornea was detected by immunofluorescence staining and myeloperoxidase.The effect of maresin1 on A.fumigatus stimulated mouse macrophage RAW264.7 cells was assessed via PCR and Western blot.Results: 1.Maresin1 inhibits the inflammatory response caused by Aspergillus fumigatus in mouse compared with control group and clinical score was also decreased(p<0.05).The m RNA and protein levels of CXCL-1 were reduced at 3d(p<0.05)and 5d(p<0.05)post infection in Maresin1 group.The m RNA and protein levels of IL-10 were enhanced at 1d,3d and 5d post infection in Maresin1 group.2.Maresin1 inhibits the MPO numbers in the Maresin1 group.The results of immunofluorescent staining shows the infiltration of neutrophils were reduced in corneal stroma in Maresin1 group.3.Maresin1 inhibits the m RNA and protein levels of CXCL-1 in 75% ethanol-killed A.fumigatus.In the meantime,maresin1 inhibits the m RNA and protein levels of CXCL-1 in 75% ethanol-killed A.fumigatus.4.Protein levels of p-GSK3? were enhanced by pretreated with Maresin1.The combination of maresin1 and Li Cl can reduced CXCL-1 m RNA levels more when compared with Li Cl group and Maresin1 group.Similarly,The combination of maresin1 and Li Cl can enhanced IL-10 m RNA levels more when compared with Li Cl group and Maresin1 group.Conclusions: These findings demonstrate that treatment with maresin1 moderates corneal inflammation through reducing neutrophil recruitment and levels of the chemokine CXCL1 and enhancing the anti-inflammatory cytokine IL-10 in A.fumigatus keratitis.Additionally,maresin1 alters levels of GSK3? phosphorylation to regulate CXCL1 and IL-10 expression in response to A.fumigatus infection.Topical administration of maresin1 may emerge as a novel anti-inflammatory molecule and has a protective role in A.fumigatus keratitis.