Expression of occludin in corneal epithelium after flap-free epipolis laser in-situ keratomileusis and laser-assisted subepithelial keratectomy

Abstract

Background Tight junctions are thought to play a significant role in the maintenance of the corneal epithelial defense for the eye,and the restoration of the tight junctions is critical during epithelial wound healing after refractive surgery.However,there are few reports about this study. Objective The aim of this study was to investigate and compare the expression of occludin in corneal epithelium following flap-free epipolis laser insitu keratomileusis (Epi-LASIK) and laser-assisted subepithelial keratectomy ( LASEK ). Methods Forty-eight clean New Zealand white rabbits were randomized into 2 groups,and 24 rabbits for each group.Flap-free Epi-LASIK and LASEK were performed in the right eyes of the rabbits in two groups,and other 2 age matched normal rabbits were as normal controls.The animals were sacrificed and the corneal samples were obtained at 1,2,3,5 days after surgery.Immunofluorescence staining was used to detect the expression of occludin in the corneal epithelium,and RT-PCR was used to identify the level of occludin mRNA in the central cornea. Results Occludin protein was expressed innormal corneal epithelium and showed the green fluorescence with the regular arrangement.The fluorescence intensity was lower in 1 -2 days in LASEK group with the irregular arrangement;while the fluorescence signal in corneal epithelium was stronger in flap-free Epi-LASIK group.3-5 days after surgery,the fluorescence intensities were bothenhanced in two groups.RT-PCR showed that the relative expression of occludin mRNA in corneal epithelium was 0.11 ±0.02,0.25 ± 0.03,0.43 ± 0.04 in LASEK group 1,2,3 days after surgery,respectively,and was lower than those of flap-free Epi-LASIK group ( 0.20± 0.04,0.44 ± 0.04,0.76 ± 0.04 ),showing significant differences between these two groups ( t =6.476,12.898,17.315,P＜ 0.05 ).No significant difference was seen in the expression of occludin mRNA in 5 days after surgery between two groups( t=-0.733,P＞0.05).The relative values of occludin mRNA expression in corneal epithelium were gradually increased with time prolongation,presenting a significant difference among various time points ( Ftime =768.903,P =0.000). Conclusions The reformation of occludin in flap-free Epi-LASIK group is faster than that in LASEK group.Therefore,flap-free Epi-LASIK is prominant in reducing the stimulated symptoms and complication after the surgery. Key words: Flap-free epipolis laser in-situ keratomileusis; Laser-assisted subepithelial keratectomy; Tight junction ; Occludin