title: Protective effect of ultra low molecular weight heparin on glutamate-induced apoptosis in cortical cells.
abstract: OBJECTIVE
  To investigate the effect of ultra low molecular weight heparin (ULMWH) on glutamate induced apoptosis in rat cortical cells and to explore the possible mechanisms.
  
  
  METHODS
  Cell viability was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptosis was first analyzed with Hoechst 33258 and then confirmed by DNA fragmentation. The concentration of free intracellular calcium ([Ca(2+)](i)) was determined with fura-2/AM fluorometry. The expression of Bcl-2 family protein and caspase-3 were evaluated with Western blot.
  
  
  RESULTS
  Typical apoptotic morphological change in rat cortical cells treated with 100 micromol/L glutamate for 24h was detected by Hoechst 33258 staining, which was then confirmed by the DNA ladder of agarose gel electrophoresis. The apoptotic rate of the glutamate treated cells was up to 33.21%, and 24 h of treatment with glutamate increased [Ca(2+)](i), down-regulated Bcl-2 expression, up-regulated Bax expression, and increased caspase-3 activation in rat cortical cells. Our research demonstrated that ULMWH pretreatment can prevent the glutamate-induced apoptosis, attenuate the increase of [Ca(2+)](i) not only in medium containing Ca(2+) but also in Ca(2+)-free medium, up-regulate the expression of Bcl-2, down-regulate the expression of Bax, and decrease caspase-3 activation.
  
  
  CONCLUSIONS
  ULMWH has neuroprotective capacity to antagonize glutamate-induced apoptosis in cortical cells, through decrease of Ca(2+) release and modulation of apoptotic processes.
authors.affiliation: Pharmacological Institute of New Drugs, School of Pharmacy, Shandong University, 44 Wen Hua Xi Road, Jinan 250012, PR China.
authors.forename: Tian-Gui
authors.initials: TG
authors.lastname: Yu
authors.forename: Qing-Zhu
authors.initials: QZ
authors.lastname: Zhang
authors.forename: Zhi-Guo
authors.initials: ZG
authors.lastname: Zhang
authors.forename: Wei-Wei
authors.initials: WW
authors.lastname: Wang
authors.forename: Sheng-Li
authors.initials: SL
authors.lastname: Ji
authors.forename: Guan-Hua
authors.initials: GH
authors.lastname: Du
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D053938:DNA Fragmentation / Q000187:drug effects; D018698:Glutamic Acid / Q000494:pharmacology*; D006495:Heparin, Low-Molecular-Weight / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D051028:bcl-2-Associated X Protein / Q000378:metabolism
chemical_list: D006495:Heparin, Low-Molecular-Weight; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D018698:Glutamic Acid; D053148:Caspase 3; D002118:Calcium