Mechanism Of MicroRNA-17 Inhibiting The Repair Of Corneal Epithelial Injury

PurposeTo observe miR-17 in C57BL/6 mice corneal epithelial damage in the process of repairing corneal epithelial healing, and to explore its mechanism by(C57BL/6 mice experiment in vivo and in vitro experimental murine corneal epithelial progenitor cell lines. TKE2 cells). MethedsThe normal C57BL/6 mice were selected to establish the corneal epithelial wound healing model and administrated with subconjunctival injection.All Experiments were divided into control group(subconjunctival injection of NTC) and experimental group(subconjunctival injection of miR-17, FN drops). The healing rate of mice corneal epithelial wound was observed in the control group(NTC) and experimental group(miR-17);The healing rate of mice corneal epithelial wound was observed in the control group(NTC) and experimental group(FN, miR-17+FN);The expression level of mice corneal limbal stem cell related genes were detected in the control group(NTC) and the experimental group(FN, miR-17+FN).Mouse corneal epithelial stem/progenitor cell line(TKE2 cells) were cultured in vitro. The experiments were divided into control group(NTC) and experimental group(miR-17, miR-17+FN, FN).The expression level of stem cell related genes were detected in control group(NTC) and experimental group(miR-17); The cell cycle were detected in control group(NTC) and experimental group(miR-17);The expression level of FN in control group(NTC) and experimental group(miR-17); The cell migration and adhesion were tested in control group(NTC) and experimental group(miR-17, miR-17+FN, FN). ResultThe corneal epithelial wound healing model were sdtablished with normal C57BL/6 mice and received subconjunctival injection of NTC,miR-17. Research results showed:to be compared with mice with subconjunctival injection of the NTC,the rate of corneal epithelial wound healing of group mice with subconjunctival injection of mi R-17 decreased significantly(P<0.05),FN drops can promote mice corneal epithelial wound healing rate(P<0.05); Immunofluorescence staining showed delta-NP63 and Ki67 showed no significant difference.The result of the study of the cultivation of TKE2 in vitro showed that compared NTC with miR-17, the expression level of the stem cell related genes(delta-NP63, Ki67, ABCG2, Bmi1) showed no significant difference(P>0.05); To be compared with NTC, miR-17 had no obvious impact on cell cycle(P>0.05); Comparing with NTC, miR-17 can inhibit the expression of FN(P<0.05).Immunofluorescence staining showed significant differences on FN expression;Comparing with NTC, mi R-17 can inhibit cell migration and adhesion(P<0.05);To be compared with miR-17,miR-17+FN significantly promote cell migration and adhesion(P<0.05);FN can promote cell migration and adhesion more obviously(P<0.05). ConclusionmiR-17 inhibite mice corneal epithelial wound healing;miR-17 inhibite mice corneal epithelial adhesion and migration;miR-17 inhibits corneal epithelial wound healing related to regulation of the expression of fibronectin.