High glucose concentration induces elevated expression of anti-oxidant and proteolytic enzymes in cultured human retinal pigment epithelial cells
Autor: | Hidetoshi Tsukamoto, Hiromu K. Mishima, Ken Yamane, Tomoko Yokoyama, Atsushi Minamoto, Shunsuke Izumi, George Hoppe, Jonathan E. Sears, Hidetoshi Yamashita |
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Rok vydání: | 2006 |
Předmět: |
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Western Protein Disulfide-Isomerases Peptide Mapping Antioxidants Statistics Nonparametric Cathepsin B Cell Line Superoxide dismutase Cellular and Molecular Neuroscience chemistry.chemical_compound Heat shock protein Image Processing Computer-Assisted medicine Humans Electrophoresis Gel Two-Dimensional Pigment Epithelium of Eye Protein disulfide-isomerase Heat-Shock Proteins chemistry.chemical_classification Dose-Response Relationship Drug biology Superoxide Dismutase Glutathione peroxidase Proteolytic enzymes Glutathione Trypsin Molecular biology Sensory Systems Culture Media Ophthalmology Glucose chemistry Biochemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization biology.protein Peptide Hydrolases medicine.drug |
Zdroj: | Experimental Eye Research. 83:602-609 |
ISSN: | 0014-4835 |
DOI: | 10.1016/j.exer.2006.02.016 |
Popis: | We investigated the differential protein expression patterns of retinal pigment epithelial (RPE) cells exposed to increased glucose concentrations. Cultured human RPE cells (ARPE-19) were exposed for 4 days with normal blood glucose concentration (5.5 mM D-glucose), followed by exposure to either normal (5.5 mM) or high (33 mM) concentrations of D-glucose for 48h. Protein extracts of glucose-treated RPE cells were then subjected to comparative proteome analysis based on 2-D gel electrophoresis. Protein spots were visualized by silver staining. The differentially expressed proteins were excised and digested in-gel with trypsin, then analysed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The expression levels of cathepsin B, glutathione peroxidase and heat shock protein 27 were increased, and that of protein disulfide isomerase decreased in high glucose treated RPE compared to normal glucose. The isoelectric point of copper/zinc-containing superoxide dismutase (Cu/Zn-SOD) shifted toward acidic region in response to high glucose. Cu/Zn-SOD activity in high glucose group was significantly lower than that in normal glucose group (P |
Databáze: | OpenAIRE |
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