title: Thiamine and benfotiamine prevent increased apoptosis in endothelial cells and pericytes cultured in high glucose.
abstract: BACKGROUND
  High glucose induces pathological alterations in small and large vessels, possibly through increased formation of AGE, activation of aldose reductase and protein kinase C, and increased flux through the hexosamine pathway. We showed previously that thiamine and benfotiamine correct delayed replication and increase lactate production in endothelial cells subjected to high glucose. We now aim at verifying the effects of thiamine and benfotiamine on cell cycle, apoptosis, and expression of adhesion molecules in endothelial cells and pericytes, under high ambient glucose.
  
  
  METHODS
  Human umbilical vein endothelial cells and bovine retinal pericytes were cultured in normal (5.6 mmol/L) or high (28 mmol/L) glucose, with or without thiamine or benfotiamine, 50 or 100 micro mol/L. Apoptosis was determined by two separate ELISA methods, measuring DNA fragmentation and caspase-3 activity, respectively. Cell cycle and integrin subunits alpha3, alpha5, and beta1 concentration were measured by flow cytometry.
  
  
  RESULTS
  Apoptosis was increased in high glucose after 3 days of culture, both in endothelium and pericytes. Thiamine and benfotiamine reversed such effects. Neither cell cycle traversal nor integrin concentrations were modified in these experimental conditions.
  
  
  CONCLUSIONS
  Thiamine and benfotiamine correct increased apoptosis due to high glucose in cultured vascular cells. Further elucidations of the mechanisms through which they work could help set the basis for clinical use of this vitamin in the prevention and/or treatment of diabetic microangiopathy.
authors.affiliation: Department of Internal Medicine, University of Turin, Italy. elena.beltramo@unito.it
authors.forename: E
authors.initials: E
authors.lastname: Beltramo
authors.forename: E
authors.initials: E
authors.lastname: Berrone
authors.forename: S
authors.initials: S
authors.lastname: Buttiglieri
authors.forename: M
authors.initials: M
authors.lastname: Porta
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods; D002453:Cell Cycle / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D020286:Pericytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D012160:Retina / Q000166:cytology* / Q000187:drug effects; D013831:Thiamine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D014471:Umbilical Veins
chemical_list: D005947:Glucose; D013831:Thiamine; C013835:benphothiamine