title: Induction of glycation suppresses glucokinase gene expression in HIT-T15 cells.
abstract: OBJECTIVE
  Chronic hyperglycaemia in patients with Type II (non-insulin-dependent) diabetes mellitus often leads to a decline in glucose-responsive insulin secretion from pancreatic beta cells, a phenomenon called glucose toxicity. Upon hyperglycaemia, glycation reaction occurs in the beta cells and induces oxidative stress. To understand the molecular basis of the beta-cell glucose toxicity, we investigated the possible effects of glycation on the expression and enzymatic activity of glucokinase, which plays a crucial part in glucose-responsive insulin secretion.
  
  
  METHODS
  Glycation and reactive oxygen species were induced in HIT-T15 cells by treatment with D-ribose and effects on glucokinase gene transcription, glucokinase protein amount, glucose phosphorylation activity, and DNA-binding activities of putative glucokinase gene transcription factors were evaluated.
  
  
  RESULTS
  When glycation was induced in HIT-T15 cells, the activity of the human glucokinase gene beta-cell-type promoter was suppressed substantially (83% reduction at 60 mmol/l D-ribose). Also, similar reductions in mRNA and protein amounts of glucokinase and in the Vmax of its enzymatic activity were observed. In agreement with the reduction in the promoter activity, the two major transcription factors of the glucokinase gene, the Pal-binding factor and PDX-1, reduced their binding to their target sequences in the glucokinase gene promoter in glycation-induced HIT cells. Because these effects of D-ribose were counteracted by aminoguanidine or N-acetylcysteine, reactive oxygen species, generated by the glycation reaction, appears to be involved in the phenomena.
  
  
  CONCLUSIONS
  The induction of the glycation reaction, which is known to occur in pancreatic beta cells in chronic hyperglycaemia, suppresses the glucokinase gene transcription and its enzymatic activity. Thus, hyperglycaemia-dependent inhibition of glucokinase activity could in part explain beta-cell glucose toxicity.
authors.affiliation: Department of Internal Medicine and Therapeutics, Osaka University Graduate School of Medicine, Suita, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Kajimoto
authors.forename: T
authors.initials: T
authors.lastname: Matsuoka
authors.forename: H
authors.initials: H
authors.lastname: Kaneto
authors.forename: H
authors.initials: H
authors.lastname: Watada
authors.forename: Y
authors.initials: Y
authors.lastname: Fujitani
authors.forename: M
authors.initials: M
authors.lastname: Kishimoto
authors.forename: K
authors.initials: K
authors.lastname: Sakamoto
authors.forename: M
authors.initials: M
authors.lastname: Matsuhisa
authors.forename: R
authors.initials: R
authors.lastname: Kawamori
authors.forename: Y
authors.initials: Y
authors.lastname: Yamasaki
authors.forename: M
authors.initials: M
authors.lastname: Hori
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D015870:Gene Expression; D005941:Glucokinase / Q000235:genetics* / Q000378:metabolism*; D006031:Glycosylation; D006146:Guanidines / Q000494:pharmacology; D018398:Homeodomain Proteins; D006801:Humans; D006943:Hyperglycemia / Q000378:metabolism; D007515:Islets of Langerhans / Q000201:enzymology*; D051266:Nuclear Respiratory Factor 1; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D012266:Ribose / Q000494:pharmacology; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D004268:DNA-Binding Proteins; D006146:Guanidines; D018398:Homeodomain Proteins; D051266:Nuclear Respiratory Factor 1; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D015534:Trans-Activators; D014157:Transcription Factors; C088388:alpha-Pal transcription factor; C084010:pancreatic and duodenal homeobox 1 protein; D012266:Ribose; D004247:DNA; D005941:Glucokinase; C004479:pimagedine; D000111:Acetylcysteine