The effects of leech percolate on cell dividing cycle of human retinal pigment epithelium cells induced by thrombin.

Abstract

Objective To investigate the effects of leech percolate on cell division cycle of culturedhuman retinal pigment epithelium (hRPE)cells induced by thrombin.Exploring mechanism of leech percolateon preventing and curing PVR.Methods (1)Human retinal pigment epithelium(hRPE)cells were cultured invitro with digest culture method. (2)The selected concentrations of leech percolate (128mg/ml,64mg/ml,32mg/ml,16mg/ml and 8mg/ml)were based on the previous experiments.With or without 0.5NIHU/mlthrombin, hRPE cells were treated with leech percolate by various concentrations for 24hrs, and the effects ofcell division cycle of leech percolate were studied with Flow Cytometer (FCM). (3)With or without0.5NIHU/ml thrombin, hRPE cells were treated with 64mg/ml leech percolate at different times(6hr, 12hr,24hrand 48hr), and the effects of cell division cycle of leech percolate were treated with FCM.Results (1)Therewas a general tendency that leech percolate of the selected concentrations arrested hRPE cells on G0/G1 phaseand 64mg/mi leech percolate inhibited 77.9% hRPE cells on G0/G1 phase and made hRPE cells on S phaseless especially. (2)During the period of 6hrs and 48hrs the longer the treat time of 64mg/ml leech percolate, themore the RPE cells arrested on G0/G1 phase.Conclusion The results approve leech percolate can inhibitproliferation of human RPE cells cultured in vitro.Its mechanism may be that leech percolate could act on celldivision cycle and arrest RPE cells on G0/G1 phase to make proliferation power of hRPE cells lower. Key words: Human retinal pigment epithelium cells; Leech percolate; Thrombin; Flow Cytometer; Cell division cycle