Leech Leachate Thrombin-induced Human Retinal Pigment Epithelial Cell Cycle

PurposeTo investigate the effects of leech percolate on cell division cycle of cultured human retinal pigment epithelium (hRPE) cells induced by thrombin. Exploring mechanism of leech percolate on preventing and curing PVR.Methods(1) Human retinal pigment epithelium (hRPE) cells were cultured in vitro with digest culture method.(2) The selected concentrations of leech percolate (128mg/mk 64mg/ml, 32mg/ml, 16mg/ml and 8mg/ml) were based on the previous experiments. With or without 0.5NIHU/ml thrombin, hRPE cells were treated with leech percolate by various concentrations for 24hrs, and the effects of cell division cycle of leech percolate were studied with Flow Cytometer(FCM) .(3) With or without 0.5NIHU/ml thrombin, hRPE cells were treated with 64mg/ml leech percolate at different times (6hr, 12hr, 24hr and 48hr) , and the effects of cell division cycle of leech percolate were treated with FCM.Results(1) There was a general tendency that leech percolate of the selected concentrations arrested hRPE cells on G₀/G₁ phase and 64mg/ml leech percolate inhibited 77.9% hRPE cells on G₀/G₁ phase and made hRPE cells on S phase less especially.(2) During the period of 6hrs and 48hrs the longer the treat time of 64mg/ml leech percolate , the more the RPE cells arrested on G₀/G₁ phase.ConclusionThe results approve leech percolate can inhibit proliferation of human RPE cells cultured in vitro. Its mechanism may be that leech percolate could act on cell division cycle and arrest RPE cells on G₀/G₁ phase to make proliferation power of hRPE cells lower.