Leech Extract On Human Retinal Pigment Epithelial Cell Thrombin Receptor-1 Expression And Leech Extract On Human Retinal Pigment Epithelial Cell Transdifferentiation

Objectiveâ‘ To research the influence of leech extract on cultured human retinal pigment epithelial (hRPE) cells' thrombin receptor-1 that is induced by thrombin.â‘¡To observe the influence of leech extract on cultured human retinal pigment epithelial (hRPE) cells' transdifferentiation.Methods(1) Human retinal pigment epithelium(hRPE)cells were cultured in vitro with digest culture method.(2) Based on the previous experiments, the selected concentrations of leech extract is 64mg/ml. 0.5NIHU/ml thrombin as induced concentration. Flourescence expression of PAR-1 was measured by immumofluorescence method. Set blank, thrombin, leech extract and add two drug at equal pace, hRPE cells were treated with three methods for 24h, to observe flourescence expression of PAR-1 on hRPE.(3) After hRPE cells were treated with 64mg/ml leech extract for 24h and 48h, the cells were immunostained with NSE and CK. To observe the influence of leech extract on hRPE cells' transdifferentiation.Results(1) Flourescence integral optical density(IOD)of PAR-1 display: thrombin group compared with normal control group, IOD decreased(p<0.01); leech extract group compared with normal control group, IOD increased(p<0.01); added leech extract and thrombin at equal pace compared with thrombin group, IOD significant increased(p<0.01).(2) After hRPE cells were treated with 64mg/ml leech extract for 24h and 48h, we found NSE positive staining on some cells, and this cells have synapse and vesicular nucleus that are neuroepithelial cells' characteristic. Conclusion(1) leech extract can inhibit PAR-1 activation on hRPE cells induced by thrombin.(2) To some extent, the experimental result confirm leech extract can induce hRPE cells transformation to neuroepithelial cells.