Inhibitory Effect Of Disintegrin On Cultured Human Lens Epithelial Cells In Vitro

OBJECTIVE To study the inhibitory effect of disintegrin on the growth of cultured human lens epithelial cells(HLECs) in vitro.METHODS In the congenital cataract surgery,the anterior capsule membrane sampled by CCC technique was cultured and subcultured and the second generations of HLECs were used in this experiment. Anti-adhesion experiment:The two passage HLECs suspended in 1640 medium were pre-incubated for 30 minutes in the presence of various concentrations of disintegin(0ng/ml,5ng/ml,10ng/ml,20ng/ml,40ng/ml,80ng/ml) at 37℃,5%CO2,then inoculated in 96-well plate coated with rat-tail collagen.After incubation for 8 hours,non-adherent cells were discarded and adherent cells were washed twice with HBSS,then the absorbance was measured at 570nm with an ELISA reader to detect the number of cells adhering.Anti-migration experiment:A denuded area was made when the second passage of HLECs was confluent into a monolayer.Cells were then exposed to media with/without 80ng/ml of disintegrin at 37℃,5%CO2,After 24hr,48hr and 72hr, the cells having migrated into the denuded area were enumerated under light microscope. Anti-proliferative experiment:16hr after the second passage of HLECs was incubated in the 96-well plate coated with rat-tail collagen at 37℃,5%CO2,they were exposed to disintegrin with different concentrations(0ng/ml,5ng/ml,10ng/ml,20ng/ml,40ng/ml,80ng/ml). Seven days later,the number of cells was determined by MTT method.RESULTS Disintegrin inhibited HLECs from attaching to collagen in a dose-dependent manner.The migration of HLECs was blocked efficiently with 80ng/ml of disintegrin.The inhibitory effect of disintegrin on HLECs proliferation was dose-dependent and a 7-day treatment with different concentrations of disintegrin significantly inhibited the proliferation of HLECs.CONCLUSIONS In vitro, disintegin can inhibit the growth of HLECs.