Role and mechanism of NTR p75 receptor in oxidative damage of retinal pigment epithelium cells / 国际眼科杂志(Guoji Yanke Zazhi)

ABSTRACT

AIM:To study the oxidative damage and mechanism of p75 NTR receptor in retinal pigment epithelial cells (RPE).METHODS:The NTR p75 receptor was used to transfer the retinal pigment epithelium cells as the experimental group,and the non transfected retinal pigment epithelium cells were used as control group.The BrdU test detected the proliferation of two groups of cells.The rate of apoptosis in two sets of apoptosis was measured by PI/in the V-FITC double dye method.The laser microscope detects the ROS levels within the cell.The flow cytometer detected the levels of ROS,mitochondrial markers,cytochrome C expression in RPE cells.The Western blot method detected the expression level of Fas,Caspase-3,and VEGF165 in RPE cells.RESULTS:The RPE cell proliferation activity was gradually decreasing (P＜0.05) with the extension of the p75 NTR receptor transfer time in experimental group.The RPE cell proliferation activity in each transfection point was significantly lower in experimental group than in the control group (P＜0.05).The percentage of RPE apoptosis was gradually increased with the extension of transfection time in experimental group(P＜0.01).The percentage of RPE cell apoptosis in the experimental group was significantly higher than the control group (P＜0.01).ROS fluorescence was significantly better in the experimental group than the control group.Flow cytometry instrument method,according to the results of the experimental group PRE ROS levels in the cell,cytochrome C was significantly higher than control group (P ＜ 0.01),RPE cell mitochondria marker levels significantly lower than the control group (P＜0.01).The results of the Western blot method showed that the expression levels of VEGF165,Fas and Caspase-3 were significantly higher in the experimental group than in the control group (P＜0.01).CONCLUSION:The over expression of p75 NTR receptor could lead to damage of mitochondria in retinal pigment epithelium cells,but it could also promote the apoptosis reaction,eventually it led to the formation of choroidal neovascularization,so it could be speculated that p75 NTR receptor is the damage factors of retinal pigment epithelium.