Diabetologie und Stoffwechsel 2021; 16(S 01): S49
DOI: 10.1055/s-0041-1727459
07. Diabeteskomplikationen/Begleiterkrankungen

Proteomic analysis of hyperglycemic effects in GLUT4-overexpressing rat cardiomyoblasts – short vs. long-term hyperglycemia

B Stratmann
1   Herz-und Diabeteszentrum NRW, Diabeteszentrum, Bad Oeynhausen, Germany
,
B Eggers
2   Ruhr-Universität Bochum, Medizinisches Proteom-Center, Bochum, Germany
,
Y Mattern
3   Herz-und Diabeteszentrum NRW, Ruhr-Universität Bochum, Diabeteszentrum, Bad Oeynhausen, Germany
,
K Marcus
2   Ruhr-Universität Bochum, Medizinisches Proteom-Center, Bochum, Germany
,
D Tschöpe
3   Herz-und Diabeteszentrum NRW, Ruhr-Universität Bochum, Diabeteszentrum, Bad Oeynhausen, Germany
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    Diabetic cardiomyopathy is characterized by oversupply of nutrients with loss of metabolic flexibility. We established a stably GLUT4 overexpressing cell line (H9C2-GLUT4) derived from H9C2 presenting a diabetic cardiomyopathy like phenotype to evaluate short and long-term hyperglycemia effects. Protein expression patterns are analyzed by proteomic approaches.

    H9C2-GLUT4 were cultured under hyperglycemic conditions (30mM glucose) either short time (14 days, ST) or long time (> 3 months, LT). Expression profiles were compared to normoglycemic conditions (20mM glucose). Samples were prepared for mass spectrometric analysis by lysis followed by tryptic digestion. Peptides were analyzed on an Orbitrap Elite (ThermoFisher Scientific) and resulting files were taken for the identification and quantification of proteins using MaxQuant and Perseus. DAVID Bioinformatics Resources 6.8 functioned as Pathway and GO term enrichment tool for further analysis of the data set.

    2,841 proteins were quantified with 881 being significantly regulated in the whole analysis set (5% FDR; ANOVA corrected p-value). Further analysis of LT and ST hyperglycemia identified 15 significantly expressed proteins in ST H9C2-GLUT4 and 156 significantly expressed proteins in the LT H9C2-GLUT4 compared to normoglycemia (5% FDR; ANOVA corrected p-value). The comparison of ST exposure to LT exposure identified 135 significantly regulated proteins. In-depth pathway and GO term enrichment analysis support effects on metabolism, protein biosynthesis and handling, and ECM composition.

    Chronic glucose overload in cardiomyoblasts induced by GLUT4 overexpression and hyperglycemia resulted in time-dependent metabolic induced proteome profile change. This may elucidate the multifaceted metabolic effects of chronic hyperglycemia on cardiomyocyte protein profile and function.


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    Artikel online veröffentlicht:
    06. Mai 2021

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