ABSTRACT
OBJECTIVE: MiRNA has been found to have therapeutic effect on corneal damage. This paper aimed to study the effect of miR-205-3p on corneal damage induced by ultraviolet (UV) radiation. MATERIALS AND METHODS: HCE cells were exposed to UV light and transfected. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot were used to determine miRNA/mRNA and protein expression. CCK-8 assay, Edu incorporation experiment, and flow cytometry were used to separately measure cell activity, proliferation and apoptosis. LC3 puncta were researched by immunofluorescence experiment. TNF-α, IL-6 and IL-1ß levels in cells were detected by enzyme-linked immunosorbent assay (ELISA) kit. MDA, SOD, and GSH-PX levels were measured using detection kits. Reactive oxygen species (ROS) level was reflected by detecting DCFH-DA density. Luciferase activity assay was performed to verify the regulating relationship between miR-205-3p and TLR4. RESULTS: UV radiation decreased HCE cell viability, proliferation, and increased HCE cell apoptosis and autophagy (all p < 0.01). When exposed UV radiation, the overexpression of miR-205-3p group elevated HCE cells viability, proliferation and weakened HCE cells apoptosis and autophagy (all p < 0.01). MiR-205-3p inhibited inflammation and oxidative stress in HCE cells induced by UV radiation (p < 0.01). MiR-205-3p directly inhibited TLR4 expression. The upregulation of TLR4 significantly reversed the effects of miR-205-3p on HCE cells phenotypes induced by UV radiation (p < 0.01). CONCLUSIONS: MiR-205-3p protected HCE cells from UV damage by inhibiting autophagy via targeting TLR4.
Subject(s)
Autophagy/physiology , Cornea/metabolism , MicroRNAs/biosynthesis , NF-kappa B/biosynthesis , Toll-Like Receptor 4/biosynthesis , Ultraviolet Rays/adverse effects , Autophagy/radiation effects , Cell Line , Cell Survival/physiology , Cell Survival/radiation effects , Cornea/pathology , Cornea/radiation effects , Epithelial Cells/metabolism , Epithelial Cells/pathology , Epithelial Cells/radiation effects , Humans , NF-kappa B/antagonists & inhibitors , Signal Transduction/physiology , Signal Transduction/radiation effects , Toll-Like Receptor 4/antagonists & inhibitorsABSTRACT
The polycarbonate sheet was modified with ammonia gaseous plasma and characterized by the contact angle measurement and ESCA analysis. The contact angles decreased significantly from 77 degrees to about 20 degrees-40 degrees, indicate that the polycarbonate sheet become more hydrophilic after plasma treatment. The ESCA analysis results showed that the hydrophilicity was mainly derived from the amino groups on the modified surface. In this study, a flow-chamber system was also constructed to evaluate the 3T3 fibroblast cells attachment phenomena on these modified sheets. Before the experimental run, the parameters of inoculated cell number and cell passage were examined previously. The results revealed that these two parameters are independent in shear experiment. And besides, 3-hours plating time has the better adhering fraction. The experimental results showed that the 3T3 fibroblast cells adhesion strength increased significantly on the plasma modified sheet.
