Effects Of IL-17 On Choroidal Endothelial Cells Proliferation, Migration And Tube Formation

Objective:To investigate the influence of IL-17 on human choroidal endothelial cells proliferation, migration, tube formation.Methods:1. Human choroidal endothelial cells were obtained using a methods of CD31 coated immuomagnetic besds combine with traditional collagenase digestion and filtration. Human choroidal endothelial cells were assessed by cell morphology with a phase-contrast microscope. Human choroidal endothelial cells were further identified by WVF with immunocytochemical staining and by CD31 with flow cytometry.2. The expression of IL-17RA and IL-17RC on isolated human choroidal endothelial cells were analyzed by flow cytometry and immunofluorescent staining.3. The pro-angiogenesis effect of IL-17 on human choroidal endothelial cells was determined by proliferation with WST-1, migration with Transwell assay and wound healing assay, and tube formation with Matrigel matrix.4. The effect of IL-17 on cytoskeleton of human choroidal endothelial cells was observed by F-actin immunofluorescent staining.Results:Human choroidal endothelial cells were isolated with a method of using CD31 coated immuomagnetic besds and identified by positive immunofluorescent staining of WVF.The expression of CD31on human choroidal endothelial cells were above 95% by flow cytometry. With those methods, human choroidal endothelial cells were successfully obtained with high purity, activity and could be passaged serially.2. High expression of IL-17RC and low expression of IL-17RA by human choroidal endothelial cells was observed by immunofluorescent staining and flow cytometry.3. A wide range of doses of human IL-17 had no direct effect on the proliferation of choroidal endothelial cells.11-17 could promoted choroidal endothelial cells migration and tube formation. The migration of choroidal endothelial cells was suppressed by incubating with IL-17 adding PI3K inhibitor. IL-17 could modulated the actin cytoskeleton reorganization in human choroidal endothelial cells and PI3K inhibitor could inhibited this process.Conclusion:IL-17 had a pro-angiogenesis effect through promoting migration and tube formation on human choroidal endothelial cells in vitro. The promoted migration effect was dependent on PI3K mediated actin cytoskeleton remodeling.