Effects of echistatin on proliferation, adhestion and migration of human lens epithelial cell in vitro

Abstract

Background The incidence of posterior capsular opacification (PCO) is increasing with the growing of cataract surgery rate.Recent researches provend that disintegrin has inhibitory effect on PCO,and echistatin is one of the disintegrin prime families.Objective This study was to investigate the effects of disintegrin and echistatin on proliferation,adhestion and migration in human lens epithelial cells (LECs) line (SRA01/04).Methods Human LECs line at logarithmic growth phase was used in the study.Cells were cocultured with medium and different concentrations of echistatin (0,2.5,5.0,7.5,10.0,15.0,20.0 mg/L) for different time.The proliferative inhibitory rates of LECs were detected by MTT method 24,48 and 72 hours after cultured.Anti-adhesion effect of echistatin were analyzed by the same assay in 90 minutes.Cell scratching test was performed to evaluate the migration ability of LECs.The width of the scratch was recorded in the culture plate covered with cells under an inverted microscope.After being cultured for 24 hours and 48 hours with echistatin,cell migration distances was examined.Results Compared with the 0 mg/L echistatin group,cells proliferation was obviously inhibited.After cultured with 2.5,5.0,7.5,10.0,15.0,20.0 mg/L echistatin,the proliferation inhibitory rate was 2.6％,15.4％,21.2％,34.7％,46.1％,58.2％ at 24 hours;6.6％,21.9％,38.2％,50.0％,60.7％,76.9％ at 48 hours and 9.8％,29.0％,46.6％,63.4％,69.1％,92.4％ at 72 hours,respectively.The absorbance value (A) in the 5.0,7.5,10.0,15.0,20.0 mg/L groups were significantly lower than that in the 0 mg/L group (P＜ 0.05).With the prolongation of acting time of Ecs,the A value of the cells was gradually reduced,with statistically significant difference (P＜0.05).The adhesion inhibitory rate was 2.6％,15.0％,26.1％,35.3％,45.2％ and 54.5％ in the 2.5,5.0,7.5,10.0,15.0,20.0 mg/L group,respectively.Compared with the result in the 0 mg/L group,the A value in the 5.0,7.5,10.0,15.0,20.0 mg/L group was statistically significant (P＜0.05).After cultured for 24 hours and 48 hours,cell migration distance shortened in the 5.0,7.5,10.0,15.0,20.0 mg/L group,showing a statistically significant difference among them (P＜0.05).Cell migration distance was gradually shortened with the lapse of action time of Ecs with the significant difference (P ＜ 0.05).Conclusions echistatin has inhibitory effects on proliferation,adhestion and migration for human LECs in vitro in time-and dose-dependent manner.It is inferred that echistatin may play a role in the prevention and treatment of PCO. Key words: Posterior capsular opacification; Disintegrin; Lens epithelial cell; Echistatin