The Role Of Dectin-1/Raf-1 Signal Cascade In Innate Responses Of Aspergillus Fumigatus Keratitis In Human

Objective:To investigate the early expression of the v-raf-1 murine leukemia viral oncogene homolog 1(Raf-1)and its role in the innate immune response of telomerase-immertalized human corneal epithelial cells(THCE cells)in fungal keratitis.Methods:THCE cells were cultured in vitro.We divided THCE cells into control group,aspergillus fumigatus group(Af),GW5074(Raf-1 inhibitor)group and Laminarin(Dectin-1 inhibitor)group randomly.The protein expression level of p-Raf-land total Raf-1 was measured by western blot.The expression of IL-6 and IL-8 mRNA in each group was detected by Real-Time Polymerase Chain Reaction(RT-PCR).Result:Western blot result showed that total Raf-1 protein levels remain unchanged at every time point each group and p-Raf-1 had a little expression in control group and reached maximal at 30 min after fungi stimulation(p<0.05).After treatment with GW5074 and Laminarin,the expression levels of p-Raf-1 had a statistically significant decline compared with Af group(p<0.05).IL-6 and IL-8 mRNA expression were increased after stimulation with fumigatus obviously compared with control group(p<0.01).The expression levels of IL-6,IL-8 mRNA had a statistically significant decline compared with Af group following treatment with GW5074(p<0.05).Conclusion:1.Expression of Raf-1 can mediate anti fungal infections by inflammatory cytokines IL-6 and IL-8 in corneal epithelial cell.2.The fungus is to play the role of anti fungal infection through Dectin-1/Raf-1 signal pathway.