细胞凋亡
转染
膜联蛋白
流式细胞术
分子生物学
生物
达皮
细胞生物学
细胞色素c
化学
细胞培养
生物化学
遗传学
作者
Liu Hw,Mengsun Yu,J Zhang,Y-B Ren,Li Yc,Y Wang,Wang Jj
出处
期刊:PubMed
日期:2018-12-24
卷期号:32 (6): 1379-1387
被引量:1
摘要
The study aimed to investigate the role and mechanism of the p75 NTR receptor in the oxidative damage of retinal pigment epithelial cells (RPE). RPE cells transfected with the p75 NTR receptor were used as the experimental group, and the untransfected RPE cells as the control group. BrdU (5-Bromo-2-deoxyUridine) was used to detect cell proliferation activity; PI/Annexin V-FITC (fluorescein isothiocyanate, FITC) double staining was used to detect the apoptosis rate of the cells. The expression of reactive oxygen species (ROS) in cells was observed by laser microscope, and the expression of ROS, mitochondrial markers, and C expression of cytochrome in cells was detected by flow cytometry. Western blot was used to detect the Fas protein, pyrolysis Caspase-3, and expression level of the vascular endothelial growth factor 165 (VEGF165) protein. The results showed that the proliferation activity of RPE cells in the experimental group decreased gradually with the increase of transfection time, and the apoptosis rate of RPE cells in the experimental group increased gradually with the increase of transfection time, and the apoptosis rate of RPE cells at each time point was significantly higher than that of the control group. The fluorescence intensity of ROS in the experimental group was significantly stronger than that in the control group (P less than 0.01). The fluorescence intensity of cytochrome C in the RPE cells in the experimental group was significantly higher than that in the control group, while the number of positive mitochondria markers in the experimental group was less than that of the control group and the fluorescence intensity was weakened. The expression of Fas protein, Caspase-3 and VEGF165 protein in the experimental group was significantly higher than that in the control group (P less than 0.01). In conclusion, p75 NTR receptor may be a cause of oxidative damage in RPE cells. .
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