The Role And Mechanism Of IL-11 In Scarring Of Fungal Keratitis

ObjectiveThe study aims to explore the effect of interleukin 11(IL-11)in the scarring process of fungal keratitis.MethodsSixty healthy male SPF grade C57BL/6J mice without eye diseases were selected,aged 8-12 weeks.Randomly divide the cages to establish the model:the control group(CON)is a simple fungal keratitis model group:subconjunctival injection of 1xPBS solution;after successful modeling,the slit lamp was used to observe and take pictures of the ocular surface of the mice,and the severity of the corneal lesions was scored clinically at 1d,2d,4d,7d,14d,and 21d,2 mice of the control(CON)group were euthanized by cervical dislocation at each time point.After the cornea was cut out,the protein level of IL-11 at different time points was measured by enzyme-linked immunosorbent assay(ELISA).The mice were sacrificed to immunofluorescence staining their whole cornea for labeling of IL-11 receptor(IL-11R)on model 1d;three mice were sacrificed at three time points of 7d,14d and 21d,using Western blotting(Western Blotting,WB)to determine the content of ?-SMA protein in mouse cornea;real-time fluorescence quantitative polymerase chain reaction(Real Time-PCR)was used to detect the expression of target gene IL-11,?-SMA in mouse cornea.Intervention was given to establish an experimental group(Neutralizing-IL-11 antibody group,N-IL-11):a fungal keratitis model with subconjunctival injection of neutralizing IL-11 antibody.After successful modeling,at the same time point as the CON group,the slit lamp was used to observe and take pictures of the ocular surface of the mice,and the clinical scores were obtained.And 7d,14d,21d at the three time points,three mice in each group of N-IL-11 were sacrificed.The content of ?-SMA protein in the cornea of the mouse was measured by Western Blotting,and the target gene ?-SMA in the cornea of the mouse was detected by Real Time-PCR.SPSS 24.0 and GraphPad Prism 8.2 statistical software were used to analyze and plot the obtained experimental data.ResultsAfter successful modeling of mice,there was no significant difference between the corneal opacity and neovascularization scores of the CON group and the N-IL-11 group at each time point of 1d,2d,and 4d(P>0.05),the clinical scores of the N-IL-11 group were lower than that of the CON group at 7d,14d,and 21d,the difference was statistically significant(P<0.05).Comparing the corneal perforation rates of the two groups,the corneal perforation rate of the CON group was 45.83%,the N-IL-11 group was 22.92%,and the N-IL-11 group was significantly lower than the CON group,the difference was statistically significant(?2=5.587,P=0.018<0.05).After establishing a fungal keratitis model,the IL-11 protein and gene expression at each time point in the CON group increased,peaking at 2d,and then the IL-11 protein level decreased and stabilized,but at each time point was still higher than the initial level before modeling,the difference is statistically significant(P<0.05);Immunofluorescence labeling of cornea in CON group was found to express IL-11R.The expression of ?-SMA protein and genes in the CON group and the N-IL-11 group increased significantly at 7d and 14 d after fungal infection,and the expression of the N-IL-11 group was lower than that of the CON group,the difference was statistically significant(P<0.05).ConclusionIn the mouse fungal keratitis model,the cornea expresses IL-11 and IL-11 R.IL-11 can promote the expression of ?-SMA protein in the cornea of fungal keratitis mice,thereby aggravating the degree of corneal scarring of fungal keratitis,and neutralizing IL-11 reduces the degree of scarring of fungal keratitis.