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Scientific reports v.6, 2016년, pp.37737 -    SCI SCIE
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Glyoxalase 1-knockdown in human aortic endothelial cells – effect on the proteome and endothelial function estimates

Stratmann, Bernd (Herz und Diabeteszentrum NRW, Diabeteszentrum, Ruhr-Universität Bochum, Bad Oeynhausen, Germany ) ; Engelbrecht, Britta (Herz und Diabeteszentrum NRW, Diabeteszentrum, Ruhr-Universität Bochum, Bad Oeynhausen, Germany ) ; Espelage, Britta C. (Herz und Diabeteszentrum NRW, Diabeteszentrum, Ruhr-Universität Bochum, Bad Oeynhausen, Germany ) ; Klusmeier, Nadine (Herz und Diabeteszentrum NRW, Diabeteszentrum, Ruhr-Universität Bochum, Bad Oeynhausen, Germany ) ; Tiemann, Janina (Herz und Diabeteszentrum NRW, Diabeteszentrum, Ruhr-Universität Bochum, Bad Oeynhausen, Germany ) ; Gawlowski, Thomas (Herz und Diabeteszentrum NRW, Diabeteszentrum, Ruhr-Universität Bochum, Bad Oeynhausen, Germany ) ; Mattern, Yvonne (Herz und Diabeteszentrum NRW, Diabeteszentrum, Ruhr-Universität Bochum, Bad Oeynhausen, Germany ) ; Eisenacher, Martin (Medical Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany ) ; Meyer, Helmut E. (Medical Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany ) ; Rabbani, Naila (Warwick Medical School, Clinical Sciences Research Laboratories, University of Warwick, University Hospital, Coventry UK ) ; Thornalley, Paul J. (Warwick Medi ) ; Tschoepe, Diethelm ; Poschmann, Gereon ; Stühler, Kai ;
  • 초록  

    Methylglyoxal (MG), an arginine-directed glycating agent, is implicated in diabetic late complications. MG is detoxified by glyoxalase 1 (GLO1) of the cytosolic glyoxalase system. The aim was to investigate the effects of MG accumulation by GLO1-knockdown under hyperglycaemic conditions in human aortic endothelial cells (HAECs) hypothesizing that the accumulation of MG accounts for the deleterious effects on vascular function. SiRNA-mediated knockdown of GLO1 was performed and MG concentrations were determined. The impact of MG on the cell proteome and targets of MG glycation was analysed, and confirmed by Western blotting. Markers of endothelial function and apoptosis were assessed. Collagen content was assayed in cell culture supernatant. GLO1-knockdown increased MG concentration in cells and culture medium. This was associated with a differential abundance of cytoskeleton stabilisation proteins, intermediate filaments and proteins involved in posttranslational modification of collagen. An increase in fibrillar collagens 1 and 5 was detected. The extracellular concentration of endothelin-1 was increased following GLO1-knockdown, whereas the phosphorylation and amount of eNOS was not influenced by GLO1-knockdown. The expression of ICAM-1, VCAM-1 and of MCP-1 was elevated and apoptosis was increased. MG accumulation by GLO1-knockdown provoked collagen expression, endothelial inflammation and dysfunction and apoptosis which might contribute to vascular damage.


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