Leech Extract On Bovine Retinal Pigment Epithelial Cells In The G-protein Signal Transduction Pathway

PURPOSETo observe effect of leech percolate on retinal pigment epithelium (RPE) cells cultured in vitro proliferation induced by thrombin, effect of leech percolate on G protein signal transduce of RPE cells.For exploring effect and mechanism of leech preventing and curing PVR. METHODS(1)Bovine retinal pigment epithelium cells were cultured in vitro with digest culture method.(2)The passage RPE were incubated for 24h, then exposed them to different dosage thrombin were placed and cultured for 24h. MTT method was used to detect the OD values.(3)Elected 128mg/ml leech percolate and 0.5U/ml thrombin; set blank, thrombin and percolation leech extract as control group, adopted 3 methods to add drugs (add two drugs at equal pace, added thrombin before percolation leech extract, added percolation leech extract before thrombin); choosed 20min as action time, to observe passage RPE growth condition, MTT method was used to detect OD values.(4)Selected 20min as action time, adopted 3 methods to add drugs, to determin amount of free Ca²⁺, cAMP catalyzed by per mg protein and PKC inside cells. RESULTS(1) Some dosage of thrombin could promote bovine RPE proliferation, especially 0.5NIHU/ml and 2NIHU/ml (p<0.01);...