| Objective: To study the regulatory effects of SHIP-1(SH2-containing inositol 5'-phosphatase 1)on the immune inflammation of herpes simplex keratitis,and to explore the treatment and effects of SHIP-1 on herpes simplex keratitis Impact of prognosis.Materials and methods: A 6-8 week old male inbred C57 BL / 6 mouse was selected,and a mouse herpes simplex keratitis infection model was established by corneal scratch inoculation method(scratched with a # character on the cornea and then dripped with virus solution),and observed clinical manifestations of a mouse model of herpes simplex keratitis at different times.SHIP-1 antagonist or SHIP-1 agonist intervention was given on the 14 th day after corneal herpes simplex virus infection modelling.The antagonist intervention was subconjunctival injection of 5 ? l of SHIP-1 antagonist 3 ?-Aminocholestane(aminocholestane),The control group was given subconjunctival injection of mice with the same dose of solvent.For agonist intervention,5 microliters of SHIP-1 agonist Rosaptor was injected subconjunctivally,and the control group was injected subconjunctivally with the same dose of solvent.Antagonists and agonists were followed up on the 1st,3rd,7th,and 14 th days after the intervention,and clinical scores were based on corneal ulcer area,depth,ulcer morphology,and corneal neovascularization.On the 14 th day after the intervention,the mice were killed by spinal dislocation.The cornea was taken for frozen section immunofluorescence staining to observe the CD4 + cells and SHIP-1 in the cornea.The frozen section for HE staining was used to observe the tissue morphology.The spleen and lymph nodes were subjected to flow cytometry.CD4 + cells and CD8 + cells were detected by surgery.Results: 1.On the first day after herpes simplex virus infection in mice,the corneal surface is rough,transparent,with clear iris texture,and the pupils are visible;on the third day after infection,the corneal surface is rough,part of the cornea is cloudy,the iris is visible,and the pupils cannot see clearly;infection On the seventh day after the corneal surface is rough,most of the cornea is cloudy,and the density is increased,and it is grayish-white turbid,part of the iris is visible,pupils can not be seen clearly,and new blood vessels are visible on the limbus.On the 14 th day after infection,the central corneal ulcer is white Dense,covering the entire pupil area,iris is not visible,new blood vessels grow to the pupil area,may have anterior chamber hemorrhage or anterior chamber pus.2.On the 14 th day after mouse SHIP-1 antagonist treatment,the corneal epithelium of the experimental group showed that the corneal epithelium was not smooth and intact,the corneal stroma was edema,and a large number of inflammatory cells were infiltrated;Degree of edema,a small amount of inflammation and infiltration.Immunofluorescence staining showed that the experimental group had more CD4 + cell infiltration than the control group,and SHIP-1 had less expression in the corneal subepithelium and stroma.Specific antagonism of SHIP-1 can aggravates the inflammatory response of herpes simplex keratitis in mice.On the 14 th day after treatment,the results of clinical scores(8.67±1.03 vs.6.33±0.52,t=5.534,p=0.003)and corneal neovascularization scores(7.83±0.41 vs.5.33±1.63,t=3.478,p=0.018)between the two groups showed significant results.Statistical difference.Flow cytometry of cervical lymph nodes and spleen of mice showed that the number of CD4 + and CD8 + cells in the experimental group was higher than that in the control group(peak 1.6K vs.675,peak 1.7K vs.550;peak 24 K vs.4K,peak 9.5K vs.1.5K).3.On the 14 th day after the SHIP-1 agonist treatment in mice,the corneal epithelium of the experimental group was smooth and intact,the corneal base was slightly edema,and the inflammatory cells were slightly infiltrated.In the control group,the corneal epithelium was smooth and intact,and the corneal stroma was slightly edema.Extensive inflammation and infiltration.Immunofluorescence staining showed that the experimental group had less CD4 + cell infiltration than the control group,and SHIP-1 was more expressed in the corneal subepithelium and stroma.After specific activation of SHIP-1,it can reduce the inflammatory response of herpes simplex keratitis in mice.On the 14 th day after treatment,the results of clinical scores(4.50±0.55 vs.6.50±0.55,t=-5.477,p=0.003)and corneal neovascularization scores(2.17±1.72 vs.3.38±0.41,t=-2.712,p=0.042)between the two groups showed significant results.Statistically significant difference.By performing flow cytometry on cervical lymph nodes in mice,the number of CD4 + and CD8 + cells in the experimental group was lower than that in the control group(peak 55 vs.140,peak 45 vs.110);Flow cytometry of mouse spleens revealed that the number of CD4 + cells in the experimental group was slightly higher than the control group(peak 275 vs.210),and the number of CD8 + cells was slightly higher than the control group(peak 160 vs.125).Conclusions: 1.The regulation of SHIP-1 can interferes with the development of herpes simplex keratitis.2.The effect of SHIP-1 on herpes simplex keratitis may be achieved through the regulation of CD4+ T cells. |
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