| Objective: The corneal epithelial injury is the most common eye injury in clinic. Consummate healing of corneal injury is important for the maintenance of normal visual function and the effect of refractive surgery. The process of corneal healing is extremely complex ,it is a combination of many cells and cytokines in coordination with time and space. Aquaporin (AQPs) is a very important protein, and CsA could palliate the inflammation of eyes, which could reducing the damage to the aquaporin because of the inflammatory reaction.This study use mechanical method to remove the corneal epithelium of mouse, and then observe the expression of AQP3 in the process of corneal epithelial healing, at the same time, we observe the impact to above- mentioned indicators after treatment with cyclosporine A eye drops in low concentration, which focuses on the mechanisms of low concentration cyclosporine A eye drops increasing the expression of AQP3, and to provide theoretical basis for studying a drug which could effectively healing the injury of corneal epithelial.Methods: (1) The impingement was marked with 2 mm diameter annular bit in the center of the corneal, and the corneal epithelial of mice was scraped with mechanical curettage method under a microscope, so the corneal epithelial damage model was established. 35 mice were randomly divided into seven groups, each group of five mice were dropped with PBS and cyclosporine A eye drops of 0.0125%,0.025%,0.05%,0.1%,0 .2% and 0.4% concentration after corneal epithelial damage. 12 hours latter, these mice were put to death, their corneal epithelial was took out and disposed with AQP3 immunohistochemical staining. (2) 60 mice were randomly divided into two groups after corneal epithelial damage, one group was dropped with PBS as the control group, and the other group was dropped with CsA eye drops of 0.05% concentration as the experimental group. After 0 h,6h,12h,18h,24h,30h, all the mice were stained with 2% fluorescein-sodium and take a picture for the eyes, and observe the corneal epithelium repair situation. Then, all the mice were put to death, corneal was took out and disposed with AQP3 immunohistochemical staining. And then, observe the AQP3 expression under the light microscope and take quantitative image analysis.Results: The expression of AQP3 can be observed in the stroma cell layer of corneal epithelial, appears brown granular, and there is no expression in the epithelial cell layer and substantia propria layer. After 12 hours of corneal epithelial damage, with the application of 0.0125%,0.025%,0.05%,0.1% concentration of cyclosporine A eye drops respectively, the expression of AQP3 after immunohistochemical staining appears a progressive trend compared with the PBS control group, but with the application of 0.2% and 0.4% concentration, there is no significant difference with the 0.1% group of the expression of AQP3.After the treatment of cyclosporine A eye drops of 0.05% concentration, the corneal epithelia can be completely recovered in 24-30 hours after corneal epithelial damage of both the control group and the experiment group. In the 6 hours after injury, there is no significant difference between the control group and the experiment group of the wound healing area (P> 0.05), but 12h,18h and 24h after injury, there were significant difference between the control group and the experimental group of the wound healing area in statistically (P <0.05).)Conclusions: AQP3 exists in the stroma cell layer of corneal epithelial. After the treatment of cyclosporine A eye drops which is below the concentration of 0.1%, the expression of AQP3 in the corneal epithelium increases, and have a good dose-effect relationship. The cyclosporin A eye drops of 0.05% concentration can speed up the reparation of corneal epithelial damage by increasing the expression of AQP3.
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