Hyperglycaemia-induced impairment of endothelium-dependent vasorelaxation in rat mesenteric arteries is mediated by intracellular methylglyoxal levels in a pathway dependent on oxidative stress

O Brouwers, P M Niessen, G Haenen, T Miyata, M Brownlee, C D Stehouwer, J G De Mey, C G Schalkwijk

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

AIMS/HYPOTHESIS: Impaired nitric oxide (NO)-dependent vasorelaxation plays a key role in the development of diabetic vascular complications. We investigated the effect of hyperglycaemia on impaired vasoreactivity and a putative role therein of the AGE precursor methylglyoxal.

METHODS: The effects of high glucose and methylglyoxal on NO-dependent vasorelaxation in isolated rat mesenteric arteries from wild-type and transgenic glyoxalase (GLO)-I (also known as GLO1) rats, i.e. the enzyme detoxifying methylglyoxal, were recorded in a wire myograph. AGE formation of the major methylglyoxal-adduct 5-hydro-5-methylimidazolone (MG-H1) was detected with an antibody against MG-H1 and quantified with ultra-performance liquid chromatography (tandem) mass spectrometry. Reactive oxygen species formation was measured with a 5-(and-6)-chloromethyl-2'7'-dichlorodihydrofluorescein diacetate acetyl ester probe and by immunohistochemistry with an antibody against nitrotyrosine.

RESULTS: High glucose and methylglyoxal exposure of mesenteric arteries significantly reduced the efficacy of NO-dependent vasorelaxation (p < 0.05). This impairment was not observed in mesenteric arteries of GLO-I transgenic rats indicating a specific intracellular methylglyoxal effect. The diabetes-induced impaired potency (pD(2)) in mesenteric arteries of wild-type rats was significantly improved by GLO-I overexpression (p < 0.05). Methylglyoxal-modified albumin did not affect NO-dependent vasorelaxation, while under the same conditions the receptor for AGE ligand S100b did (p < 0.05). Methylglyoxal treatment of arteries increased intracellular staining of MG-H1 in endothelial cells and adventitia by fivefold accompanied by an eightfold increase in the oxidative stress marker nitrotyrosine. Antioxidant pre-incubation prevented methylglyoxal-induced impairment of vasoreactivity.

CONCLUSIONS/INTERPRETATION: These data show that hyperglycaemia-induced impairment of endothelium-dependent vasorelaxation is mediated by increased intracellular methylglyoxal levels in a pathway dependent on oxidative stress.

Original languageEnglish
JournalDiabetologia
Volume53
Issue number5
Pages (from-to)989-1000
Number of pages12
ISSN0012-186X
DOIs
Publication statusPublished - May 2010
Externally publishedYes

Keywords

  • Analysis of Variance
  • Animals
  • Cell Count
  • Cell Line
  • Cells, Cultured
  • Endothelium, Vascular
  • Humans
  • Hyperglycemia
  • Immunohistochemistry
  • Lactoylglutathione Lyase
  • Mesenteric Arteries
  • Nitric Oxide
  • Oxidative Stress
  • Pyruvaldehyde
  • Rats
  • Rats, Transgenic
  • Reactive Oxygen Species
  • Vasodilation

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