title: Enhancement of homocysteine toxicity to insulin-secreting BRIN-BD11 cells in combination with alloxan.
abstract: Previous studies have shown that homocysteine (HC) has a detrimental impact on insulin secretion and pancreatic beta cell function. The aim of the present study was to determine the role of reactive oxygen species (ROS) in the in vitro toxic effects of HC on insulin secretion and function of BRIN-BD11 insulin-secreting cells. In this study, insulin secretion from BRIN-BD11 cells was determined radioimmunologically, cell viability by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and glucokinase activity by a glucose phosphorylation assay following culture with HC plus alloxan (Alx). Treatment with HC resulted in concentration-dependent inhibition of insulin secretion induced by glucose and other insulinotropic agents. HC in combination with Alx resulted in a more pronounced decline in insulin secretion, including that induced by 20  mM alanine, by 43% (P<0.001) and 30  mM KCl by 60% (P<0.001), compared with control culture. The glucokinase phosphorylating capacity in cells cultured with HC plus Alx was significantly lower, compared with control cells. The cells also displayed a significant 84% (P<0.001) decline in cell viability. Prolonged, 72-h culture of insulin-secreting cells with HC followed by 18-h culture without HC did not result in full restoration of beta cell responses to insulinotropic agents. In vitro oxygen consumption was enhanced by a combination of Alx with HC. The study arrived at the conclusion that HC generates ROS in a redox-cycling reaction with Alx that explains the decline in viability of insulin-secreting cells, leading to reduced glucokinase phosphorylating ability, diminished insulin secretory responsiveness and cell death.
authors.affiliation: Diabetes Research Group, SAAD Centre for Pharmacy and Diabetes, University of Ulster, Coleraine, Northern Ireland, UK. siobhan.scullion@liv.ac.uk
authors.forename: S M
authors.initials: SM
authors.lastname: Scullion
authors.forename: S M J
authors.initials: SM
authors.lastname: Scullion
authors.forename: E
authors.initials: E
authors.lastname: Gurgul-Convey
authors.forename: M
authors.initials: M
authors.lastname: Elsner
authors.forename: S
authors.initials: S
authors.lastname: Lenzen
authors.forename: P R
authors.initials: PR
authors.lastname: Flatt
authors.forename: N H
authors.initials: NH
authors.lastname: McClenaghan
mesh_terms: D000496:Alloxan / Q000008:administration & dosage / Q000494:pharmacology / Q000633:toxicity*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004338:Drug Combinations; D004353:Drug Evaluation, Preclinical; D004357:Drug Synergism; D005941:Glucokinase / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006710:Homocysteine / Q000008:administration & dosage / Q000494:pharmacology / Q000633:toxicity*; D006801:Humans; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D010100:Oxygen / Q000493:pharmacokinetics; D017382:Reactive Oxygen Species / Q000378:metabolism / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D004338:Drug Combinations; D007328:Insulin; D017382:Reactive Oxygen Species; D006710:Homocysteine; D000496:Alloxan; D005941:Glucokinase; D005947:Glucose; D010100:Oxygen