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Glycolytic enzyme expression and pyruvate kinase activity in cultured fibroblasts from type 1 diabetic patients with and without nephropathy

Author:
  • E Iori
  • R Millioni
  • L Puricelli
  • Giorgio Arrigoni
  • L Lenzini
  • R Trevisan
  • Peter James
  • GP Rossi
  • LA Pinna
  • P Tessari
Publishing year: 2008
Language: English
Pages: 627-633
Publication/Series: Biochimica et Biophysica Acta - Molecular Basis of Disease
Volume: 1782
Issue: 11
Document type: Journal article
Publisher: Elsevier

Abstract english

Since type 1 diabetes mellitus (T1DM) patients with nephropathy (DN+) are insulin-resistant, we aimed to identify (new) potential molecular sites involved in the alterations of glucose metabolism in these patients. We examined the expression of glycolytic enzymes in cultured fibroblasts from T1DM(DN+) patients as compared to those from T1DM patients without nephropathy (DN-) and from controls. Pyruvate kinase (PK) activity was also determined. Human skin fibroblasts were grown in normal glucose (6 mM), RNAs and proteins were analyzed, respectively, using cRNA microarray and two-dimensional electrophoresis followed by identification with mass spectrometry. PK activity was measured using a spectrophotometric assay. As compared to controls, increases in the gene expression of hexokinase, phosphoglucomutase, phosphofructokinase, aldolase and triosephosphate isomerase were found in T1DM(DN+) patients, but not in T1DM(DN-) patients. In T1DM(DN+) patients, the protein analysis showed an altered expression of three glycolytic enzymes: triosophosphate isomerase, enolase and PK. In addition, PK activity in fibroblasts from T1DM(DN+) patients was lower than that in T1DM(DN-) and in controls. In conclusion, this study reports novel alterations of enzymes involved in glucose metabolism that may be associated with the pathophysiology of insulin resistance and of renal damage in T1DM(DN+) patients.

Keywords

  • Immunology in the medical area

Other

Published
  • ISSN: 0925-4439
Peter James
E-mail: peter.james [at] immun.lth.se

Professor

Department of Immunotechnology

+46 46 222 14 96

+46 70 247 79 60

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