Shimoi, K., Okitsu, A., Green, M.H.L., Lowe, J.E., Ohta, T., Kaji, K., Terato, H., Ide, H. and Kinae, N. (2001) Oxidative DNA damage induced by high glucose and its suppression in human umbilical vein endothelial cells Mutation research - fundamental and molecular mechanisms of mutagenesis, 480. pp. 371-378. ISSN 0027-5107Full text not available from this repository.
In order to investigate the mechanism of the production of oxidative DNA damage by hyperglycemia, we measured formamidopyrimidine N-glycosylase (FPG)-sensitive sites by the comet assay in human umbilical vein endothelial cells (HUVECs) cultured under various conditions including high glucose. Mean values of FPG-sensitive sites were higher in HUVECs cultured for 5 days in high glucose (45 mM) compared with normal glucose (5 mM) medium (P<0.001). FPG-sensitive sites increased in a time-dependent manner under high glucose treatment (3 days: P<0.05, 5 days: P<0.001), whereas Image-glucose, which is taken up poorly into the cells, gave a slight increase in FPG-sensitive sites (P<0.05). Flow cytometric analysis using 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate, di(acetoxymethyl ester) showed that incubation with Image-glucose produced more reactive oxygen species than incubation with Image-glucose. However, these increases were slight (1.22- and 1.12-folds, respectively). Incubation of HUVECs with aminoguanidine (100 µM) or pyridoxamine (1 mM), which are inhibitors of glycation, decreased the levels of FPG-sensitive sites (P<0.001). However, these inhibitors did not suppress the intracellular generation of reactive oxygen species induced by high glucose. These results indicate that FPG-sensitive sites induced by high glucose are not due to intracellular reactive oxygen species. In order to clarify what caused the induction of FPG-sensitive sites, we investigated the effect of glyoxal and 3-deoxyglucosone (3-DG) on the induction of FPG-sensitive sites and the intracellular production of reactive oxygen species in HUVECs. Glyoxal and 3-DG at a concentration of 100 µg/ml induced FPG-sensitive sites (P<0.001, P<0.01, respectively). In contrast, glyoxal did not generate reactive oxygen species inside HUVECs. The results shown in this study suggest that glyoxal formed intracellularly or extracellularly during high glucose treatment might induce FPG-sensitive sites by a mechanism not involving reactive oxygen species.
|Item Type:||Journal article|
|Uncontrolled Keywords:||High glucose; Oxidative DNA damage; Comet assay; Glyoxal; Pyridoxamine|
|Subjects:||B000 Health Professions > B100 Anatomy Physiology and Pathology|
|DOI (a stable link to the resource):||10.1016/S0027-5107(01)00196-8|
|Faculties:||Faculty of Science and Engineering > School of Pharmacy and Biomolecular Sciences|
|Depositing User:||editor spbs|
|Date Deposited:||29 Nov 2006|
|Last Modified:||06 Mar 2013 15:04|
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