摘要
AIM: To investigate the protective mechanism of Gingko Biloba extract(EGb761) on the ability of retinal pigment epithelial(RPE) cells to resist light-induced damage in a comparative proteomics study. · METHODS: Human RPE cells(ARPE-19) were randomly distributed to one of three groups: normal control(NC group) and light-damaged model without or with EGb761 group(M and ME groups,respectively). The light-damaged model was formed by exposing to white light(2 200 ±300)lx for 6h. The RPE cells in ME group were conducted with EGb761(100μg/mL) before light exposure. The soluble cellular proteins extracting from each groups were separated by two-dimensional electrophoresis and stained by silver staining. Different proteins in the profiles of the gels were analyzed by Image Master Software. Two-fold expressing protein spots were identified by Matrix-assisted laser desorption/ ionization tandem time-of-flight(MALDI-TOF/TOF) mass spectrometry. ·RESULTS: NC,M and ME groups displayed 1 892±71,2 145 ±23 and 2 216 ±85 protein spots,respectively. We identified 33 proteins with different expression levels between the NC and M groups,25 proteins between the M and ME groups,and 11 proteins between the NC and ME groups. MALDI-TOF/TOF mass spectrometry successfully identified 16 proteins,including metabolic enzymes,cytoskeletal proteins,anti-oxidation proteins,and others. ·CONCLUSION: Differences in some important proteins,such as cathepsin B,heat shock protein,and cytochrome C reductase,indicated that multiple pathways may be induced in light-damaged RPE cells and the protective effect of EGb761.
AIM: To investigate the protective mechanism of Gingko Biloba extract(EGb761) on the ability of retinal pigment epithelial(RPE) cells to resist light-induced damage in a comparative proteomics study. · METHODS: Human RPE cells(ARPE-19) were randomly distributed to one of three groups: normal control(NC group) and light-damaged model without or with EGb761 group(M and ME groups,respectively). The light-damaged model was formed by exposing to white light(2 200 ±300)lx for 6h. The RPE cells in ME group were conducted with EGb761(100μg/mL) before light exposure. The soluble cellular proteins extracting from each groups were separated by two-dimensional electrophoresis and stained by silver staining. Different proteins in the profiles of the gels were analyzed by Image Master Software. Two-fold expressing protein spots were identified by Matrix-assisted laser desorption/ ionization tandem time-of-flight(MALDI-TOF/TOF) mass spectrometry. ·RESULTS: NC,M and ME groups displayed 1 892±71,2 145 ±23 and 2 216 ±85 protein spots,respectively. We identified 33 proteins with different expression levels between the NC and M groups,25 proteins between the M and ME groups,and 11 proteins between the NC and ME groups. MALDI-TOF/TOF mass spectrometry successfully identified 16 proteins,including metabolic enzymes,cytoskeletal proteins,anti-oxidation proteins,and others. ·CONCLUSION: Differences in some important proteins,such as cathepsin B,heat shock protein,and cytochrome C reductase,indicated that multiple pathways may be induced in light-damaged RPE cells and the protective effect of EGb761.
基金
Supported by National Natural Science Foundation of China(No.30500676)
